天花粉蛋白抑制乳腺癌生长及逆转ERα基因甲基化的研究
|
摘要
乳腺癌是严重危害女性健康的最常见的肿瘤,发病率在逐年上升。大量的实验与临床研究发现雌激素在乳腺癌的发生发展中起着重要作用。在乳腺癌人群中,大约2/3乳腺癌患者表达雌激素受体(Estrogen Receptor,ER),此类患者肿瘤细胞生长具有雌激素依赖性,其作用机制是雌激素通过与其受体结合,作用于乳腺上皮,促使乳腺肿瘤的发生与发展。临床上对于ER阳性乳腺癌患者常采取抗雌激素的内分泌治疗,该疗法与联合化疗疗效相当,且成本低,毒副反应小,口服方便。但是,临床上仍有1/3的原发乳腺癌患者缺乏ER表达,此类患者对内分泌治疗反应差。
ERα基因定位于染色体6q24-27上,其表达的ERα蛋白常作为一个重要分子标志物用于评估乳腺癌治疗的激素依赖性和预测内分泌治疗效果。研究发现ERα阴性乳腺癌细胞缺乏ERα蛋白的表达不是由于ERα基因内部发生突变,因此推测ERα基因的失转录是导致激素抵抗的可能机制。文献报道ERα阴性乳腺癌ER基因启动子CpG岛(富含胞嘧啶和鸟嘌呤)呈高甲基化,因为基因启动子CpG岛的甲基化可直接抑制基因转录,因此ERα蛋白的失表达很可能是由于雌激素受体(ER)基因启动子CpG岛异常的高甲基化所致。
本研究初步探讨了具有中国特色的抗肿瘤中药天花粉蛋白(Trichosanthin,TCS)对MDA-MB-231和MCF-7乳腺癌细胞系的生长抑制作用,并进一步研究其诱导雌激素受体(ER)阴性人乳腺癌细胞株MDA-MB-231的ERα基因去甲基化的可能性及ERα基因表达情况和功能状态,以期阐明乳腺癌中ERα基因的甲基化情况,以及通过改变ERα基因的甲基化而为乳腺癌治疗提供新的方法。具体实验研究设计如下:
(1)以MDA-MB-231和MCF-7乳腺癌细胞作为研究对象,TCS作用于乳腺癌细胞株,观察其对癌细胞生物学行为的影响。
(2)检测MDA-MB-231和MCF-7乳腺癌细胞中ERα的甲基化状态、ERαmRNA和蛋白表达,研究它们之间的相关性,初步探讨ERα在乳腺癌中表达沉默的机制。
(3)以TCS作用于MDA-MB-231乳腺癌细胞,并以去甲基化药物5-氮杂胞苷(5-azacytidine,5-Aza-C)为阳性对照,观察上述指标的变化及细胞对内分泌治疗药物他莫昔芬(Tamoxifen,TAM)的敏感性,探讨TCS的去甲基化效应。
(4)最后,在裸鼠体内实验中,研究TCS的抗乳腺癌效果及其在体内应用的安全性,并研究其对ERα表达的影响,以证明TCS的抑瘤作用和去甲基化在ERα表达下调中的作用。
第一章TCS对MDA-MB-231和MCF-7乳腺癌细胞的影响
目的:探讨TCS对MDA-MB-231和MCF-7乳腺癌细胞生物学行为的影响以及可能的量效关系。
方法:(1)MDA-MB-231和MCF-7乳腺癌细胞株常规培养,用TCS处理细胞,光学显微镜下观察药物处理前后乳腺癌细胞的形态学改变;(2)MTT法检测TCS处理MDA-MB-231和MCF-7乳腺癌细胞前后细胞的增殖情况;(3)TCS(90μg/ml)处理MDA-MB-231和MCF-7乳腺癌细胞48小时后,用流式细胞仪检测用药前后细胞周期时相变化和细胞凋亡率。
结果:(1)TCS处理MDA-MB-231和MCF-7乳腺癌细胞后细胞形态发生了变化,部分细胞出现变性坏死;(2)四唑盐比色法(MTT法)显示随着TCS作用时间延长,浓度增大,细胞的生长受到明显的抑制(p＜0.05);(3)流式细胞仪检测发现,TCS使MDA-MB-231和MCF-7乳腺癌细胞阻滞于G0/G1期,细胞不能通过G1/S限制点进入S期进行复制,S期细胞比例明显减少。流式细胞仪检测到凋亡峰。
结论:TCS可抑制MDA-MB-231和MCF-7乳腺癌细胞增殖,并呈时间和浓度依赖性;TCS可诱导MDA-MB-231和MCF-7乳腺癌细胞的凋亡,使细胞阻滞在G0/G1期。
第二章TCS对MDA-MB-231乳腺癌细胞ERα基因表达及甲基化的影响
目的:探讨乳腺癌ERα基因表达缺失与ERα基因高甲基化的相关性,并研究TCS对ERα阴性乳腺癌细胞株MDA-MB-231 ERα基因表达和CpG岛甲基化的影响及细胞经TCS处理后,可否恢复对TAM的敏感性。
方法:选用ERα阴性人乳腺癌细胞株MDA-MB-231为研究对象,ERα阳性人乳腺癌细胞株MCF-7为阳性对照,分别用去甲基化剂5-Aza-C(90μg/ml)和抗肿瘤中药TCS(5μM)处理MDA-MB-231乳腺癌细胞,并进行以下研究:(1)RT-PCR法检测细胞ERαmRNA的表达情况;(2)甲基化特异性PCR(methylation specific PCR,MSP)方法检测细胞ERα基因5′CpG岛甲基化的情况;(3)Western blot方法检测细胞ERα蛋白的表达;(4)MTT法检测MDA-MB-231乳腺癌细胞经TCS和5-Aza-C处理后,雌激素拮抗剂TAM对肿瘤细胞生长的影响。
结果:(1)RT-PCR法显示ERα阴性人乳腺癌细胞株MDA-MB-231ERαmRNA无表达,ERα阳性人乳腺癌细胞株MCF-7 ERαmRNA表达呈强阳性,5-Aza-C和TCS处理后的MDA-MB-231细胞ERαmRNA恢复表达;(2)MSP鉴定ERα甲基化状态:MDA-MB-231细胞仅甲基化引物扩增出特异PCR条带;MCF-7细胞仅非甲基化引物扩增出特异PCR条带;5-Aza-C和TCS处理后的MDA-MB-231细胞仅非甲基化引物扩增出特异PCR条带;(3)Western-blot方法分析显示MDA-MB-23细胞ERα蛋白不表达;MCF-7细胞ERα蛋白表达呈强阳性;5-Aza-C和TCS作用MDA-MB-231细胞后ERα蛋白恢复了表达;(4)MTT检测TCS和5-Aza-C处理ERα阴性MDA-MB-231细胞后,TAM恢复了对肿瘤细胞的生长抑制作用,且TCS处理组抑制作用比5-Aza-C组强(p＜0.05)。
结论:乳腺癌ERα基因表达缺失与ERα基因高甲基化有关,TCS可逆转ERα的甲基化,使ERαmRNA和蛋白重新表达,并可恢复对内分泌治疗药物TAM的敏感性。
第三章TCS对ERα阴性乳腺癌裸鼠移植瘤生长及ERα表达的影响
目的:研究TCS对ERα阴性乳腺癌裸鼠移植瘤生长及移植瘤内ERα基因表达和CpG岛甲基化的影响。
方法:(1)将MDA-MB-231乳腺癌细胞悬液接种于20只BALB/C(nu/nu)雌性裸鼠(SPF级)。18只裸鼠成瘤后将其分为三组:对照组、TCS组和盐水组,每组6只。分别于移植瘤旁注射TCS(0.4mg/kg)、盐水0.2ml,隔日注射一次,共7次;(2)检测裸鼠移植瘤体积和瘤重,计算肿瘤生长抑制率,检测各组裸鼠肝肾功能和血常规;(3)RT-PCR检测TCS对裸鼠乳腺癌移植瘤ERαmRNA表达的影响;(4)MSP检测TCS对裸鼠乳腺癌移植瘤ERα甲基化状态的影响;(5)Western blot检测TCS对裸鼠乳腺癌移植瘤组织ERα蛋白表达的影响。
结果:(1)细胞接种裸鼠成瘤成功率为90%(18/20);(2)TCS组裸鼠移植瘤体积和瘤重较对照组明显缩小(P＜0.05);(3)TCS对荷瘤裸鼠肾功能、血液系统无明显影响(P＞0.05),可引起谷丙转氨酶轻度升高(P＜0.05);(4)RT-PCR结果显示TCS治疗组裸鼠乳腺癌移植瘤ERαmRNA重新表达,而对照组和盐水组则无ERαmRNA表达;(5)裸鼠移植瘤模型的对照组和盐水组仅甲基化引物扩增出特异PCR条带,而TCS组仅非甲基化引物扩增出特异PCR条带;(6)治疗组乳腺癌组织可见ERα蛋白表达,对照组和盐水组ERα蛋白表达阴性。
结论:TCS可抑制乳腺癌肿瘤生长且在体内应用有一定的安全性;体内实验进一步证实TCS可以逆转ERα基因的高甲基化,使ERαmRNA和蛋白重新表达。
Breast cancer is the most common cancer in women,of which the morbidity increases year by year.Many researches presented estrogen is important in breast cancer.Two thirds of primary breast cancer have the expression of estrogen receptor(ER) and the growth of tumor cell is hormonal dependent.The function of estrogen depends on binding with its receptor and promoting the progression of breast tumor.The clinical therapy of breast cancer patients(ER positive) usually includes endocrinotherapy,which is equal to chemotherapy.However,one third breast cancer patients(ER negative) are not sensitive to endocrine therapy.
Estrogen receptor alpha(ERα) gene is located on the chromosome 6q24-27.The proteinum expressed by ERαgene is usually an important molecule marker to evaluate hormonal dependence and predicts the effect of endocrine therapy on breast cancer.Current researches report the mechanism of ERαgene expression silencing is not due to gene mutation, but probably associate with the genetic transcription.Because of the 5'CpG island methylation in the core promotor of ERαgene,the genetic transcription is suppressed which induce to the silence of ERαgene.
Trichosanthin(TCS) is a kind of traditional Chinese medicine,which we choose as experimental drug of demethylation.The purpose is to investigate the the effect of TCS on proliferation of MDA-MB-231 and MCF-7 breast cancer cell,the demethylating effect on ERαgene in MDA-MB-231 breast cancer cell,the relationship between demethylation and gene expression;meanwhile,to explore a new therapeutic strategy for breast cancer by demethylating treatment.The design of the study include:
(1) Investigate the effect of TCS acting on breast cancer cell MDA-MB-231 and MCF-7;
(2) Measure the demethylation of ERαmRNA and protein.Analyze the relationship between ERαmRNA expression and methylation;
(3) Explore the demethylating effect of TCS on MDA-MB-231 breast cancer cell and the sensitivity to endocrine therapy;
(4) Study the suppressive effect of TCS on transplanted human breast cancer in nude mice and the security in vivo.
The PartⅠ
Objective:Investigate the effects of TCS on proliferation,apoptosis of MDA-MB-231 and MCF-7 breast cancer cell.
Methods:(1) Human breast cancer cell lines MDA-MB-231 and MCF-7 were growing at 37℃and 5%CO_2.The morphological change of cells were observed by optical microscope after treated with TCS(90μg/ml) for 72 hours;(2) MTT was performed for the proliferation of MDA-MB-231 and MCF-7 cell after treated with TCS;(3) The period and apoptosis rate of MDA-MB-231 and MCF-7 cell were tested by Flow Cytometry method(FCM) after treated with TCS(90μg/ml) for 48 hours.
Results:(1) The cellular morphology changed after treated with TCS.Denaturation and necrosis occurred in some cells;(2) TCS could inhibit the growth of MDA-MB-231 and MCF-7 cell by MTT test;(3) TCS could suspend MDA-MB-231 and MCF-7 cell generation at G0/G1 period.It also induced MDA-MB-231 and MCF-7 cell apoptosis by FCM test.
Conclusion:TCS could inhibit MDA-MB-231 and MCF-7 cell proliferation in a time- and concentration-dependent manner.TCS could induce MDA-MB -231 and MCF-7 cell apoptosis and suspend the cell generation at G0/G1 period.
The PartⅡ
Objective:Explore the relationship between ERαgene expression silencing and hypermethylation.Study the effect of TCS on ERαgene expression and 5'CpG island methylation in MDA-MB-231 breast cancer cell.Test the possibility that TCS may induce MDA-MB-231 breast cancer cell sensitive to endocrine therapy.
Methods:We chose MDA-MB-231 breast cancer cell(ERαnegative) and MCF-7 breast cancer cell(ERαpositive).MDA-MB-231 cell was treated with TCS(90μg/ml) or 5-Aza-C(5μM).(1) The ERαmRNA level of the cells was measured by RT-PCR;(2) The 5'CpG island methylation of ERαgene was tested by Methylation Specific PCR(MSP) test;(3) Western blot method was performed for ERαprotein in the cells;(4) MTT method tested whether Tamoxifen could induce the growth inhibition of MDA-MB-231 cell after treated with TCS or 5-Aza-C.
Results:(1) The expression of ERαmRNA in MDA-MB-231 cell was silencing by RT-PCR test,while the expression of ERαmRNA in MCF-7 cell was positive.The re-expression of ERαmRNA in MDA-MB-231 cell treated with TCS or 5-Aza-C was shown by RT-PCR; (2) Methylation of ERαgene was found in MDA-MB-231 cell with MSP technique.Demethylation of ERαgene was found in MCF-7 cell and MDA-MB-231 cell treated with TCS or 5-Aza-C;(3) The expression of ERαprotein was negative in MDA-MB-231 cell,but positive in MCF-7 cell determined by western blot method,and re-expressed when treated with TCS or 5-Aza-C;(4) Tamoxifen could inhibit the proliferation of MDA-MB-231 cell treated with TCS efficiently through MTT method and the suppressive effect of TCS was stronger than 5-Aza-C(p＜0.05).
Conclusion:ERαgene expression silencing was related to the hypermethylation of ERαgene.TCS could induce the re-expression of ERαmRNA and protein in MDA-MB-231 cell by demethylating ERαgene.TCS could induce MDA-MB-231 breast cancer cell sensitive to Tamoxifen.
The PartⅢ
Objective:Study the suppressive effect of TCS on transplanted human breast cancer in nude mice and effect on the expression of ERα.
Methods:(1) 20 nude mice were transplanted with human breast cancer MDA-MB-231 cell to construct tumor models.Once the tumor model in nude mice was established,18 nude mice were devided into 3 groups:control group(without treatment),TCS group and saline group. TCS(0.4mg/kg) and sodium chloride were injected around the tumor.A total of 7 injections were given,once every 2 days;(2) Observations were made on tumor suppression rate,tumor volume and weight,liver and renal function,blood routine test;(3) The ERαmRNA level in transplanted neuplasma was measured by RT-PCR assays;(4) The methylation of ERαgene in transplanted neuplasma was performed by MSP test;(5) Immunohistochemistry and western blot were used to detemine the expression of ERαprotein in the 3 groups.
Results:(1) The subcutaneous tumor model in nude mice was successfully established(18/20);(2) The tumor volume and weight in TCS group decreased(p＜0.05);(3) Renal function and blood system were normal(P＞0.05) in TCS group.SGPT in TCS group was a little higher than control group(P＜0.05);(4) The re-expression of ERαmRNA was shown by RT-PCR in TCS group;(5) Demethylation of ERαgene was found in transplanted neuplasma treated with TCS by MSP test;(6) The expression,of ERαprotein was found in transplanted neuplasma treated with TCS by immunohistochemistry and western blot test.
Conclusions:TCS could effectively suppress human breast cancer cells growth rate in subcutaneous tumor of nude mice and it was relatively safe to use it.TCS could induce the re-expression of ERαmRNA and protein in vivo by demethylating ERαgene.
ERα基因定位于染色体6q24-27上,其表达的ERα蛋白常作为一个重要分子标志物用于评估乳腺癌治疗的激素依赖性和预测内分泌治疗效果。研究发现ERα阴性乳腺癌细胞缺乏ERα蛋白的表达不是由于ERα基因内部发生突变,因此推测ERα基因的失转录是导致激素抵抗的可能机制。文献报道ERα阴性乳腺癌ER基因启动子CpG岛(富含胞嘧啶和鸟嘌呤)呈高甲基化,因为基因启动子CpG岛的甲基化可直接抑制基因转录,因此ERα蛋白的失表达很可能是由于雌激素受体(ER)基因启动子CpG岛异常的高甲基化所致。
本研究初步探讨了具有中国特色的抗肿瘤中药天花粉蛋白(Trichosanthin,TCS)对MDA-MB-231和MCF-7乳腺癌细胞系的生长抑制作用,并进一步研究其诱导雌激素受体(ER)阴性人乳腺癌细胞株MDA-MB-231的ERα基因去甲基化的可能性及ERα基因表达情况和功能状态,以期阐明乳腺癌中ERα基因的甲基化情况,以及通过改变ERα基因的甲基化而为乳腺癌治疗提供新的方法。具体实验研究设计如下:
(1)以MDA-MB-231和MCF-7乳腺癌细胞作为研究对象,TCS作用于乳腺癌细胞株,观察其对癌细胞生物学行为的影响。
(2)检测MDA-MB-231和MCF-7乳腺癌细胞中ERα的甲基化状态、ERαmRNA和蛋白表达,研究它们之间的相关性,初步探讨ERα在乳腺癌中表达沉默的机制。
(3)以TCS作用于MDA-MB-231乳腺癌细胞,并以去甲基化药物5-氮杂胞苷(5-azacytidine,5-Aza-C)为阳性对照,观察上述指标的变化及细胞对内分泌治疗药物他莫昔芬(Tamoxifen,TAM)的敏感性,探讨TCS的去甲基化效应。
(4)最后,在裸鼠体内实验中,研究TCS的抗乳腺癌效果及其在体内应用的安全性,并研究其对ERα表达的影响,以证明TCS的抑瘤作用和去甲基化在ERα表达下调中的作用。
第一章TCS对MDA-MB-231和MCF-7乳腺癌细胞的影响
目的:探讨TCS对MDA-MB-231和MCF-7乳腺癌细胞生物学行为的影响以及可能的量效关系。
方法:(1)MDA-MB-231和MCF-7乳腺癌细胞株常规培养,用TCS处理细胞,光学显微镜下观察药物处理前后乳腺癌细胞的形态学改变;(2)MTT法检测TCS处理MDA-MB-231和MCF-7乳腺癌细胞前后细胞的增殖情况;(3)TCS(90μg/ml)处理MDA-MB-231和MCF-7乳腺癌细胞48小时后,用流式细胞仪检测用药前后细胞周期时相变化和细胞凋亡率。
结果:(1)TCS处理MDA-MB-231和MCF-7乳腺癌细胞后细胞形态发生了变化,部分细胞出现变性坏死;(2)四唑盐比色法(MTT法)显示随着TCS作用时间延长,浓度增大,细胞的生长受到明显的抑制(p＜0.05);(3)流式细胞仪检测发现,TCS使MDA-MB-231和MCF-7乳腺癌细胞阻滞于G0/G1期,细胞不能通过G1/S限制点进入S期进行复制,S期细胞比例明显减少。流式细胞仪检测到凋亡峰。
结论:TCS可抑制MDA-MB-231和MCF-7乳腺癌细胞增殖,并呈时间和浓度依赖性;TCS可诱导MDA-MB-231和MCF-7乳腺癌细胞的凋亡,使细胞阻滞在G0/G1期。
第二章TCS对MDA-MB-231乳腺癌细胞ERα基因表达及甲基化的影响
目的:探讨乳腺癌ERα基因表达缺失与ERα基因高甲基化的相关性,并研究TCS对ERα阴性乳腺癌细胞株MDA-MB-231 ERα基因表达和CpG岛甲基化的影响及细胞经TCS处理后,可否恢复对TAM的敏感性。
方法:选用ERα阴性人乳腺癌细胞株MDA-MB-231为研究对象,ERα阳性人乳腺癌细胞株MCF-7为阳性对照,分别用去甲基化剂5-Aza-C(90μg/ml)和抗肿瘤中药TCS(5μM)处理MDA-MB-231乳腺癌细胞,并进行以下研究:(1)RT-PCR法检测细胞ERαmRNA的表达情况;(2)甲基化特异性PCR(methylation specific PCR,MSP)方法检测细胞ERα基因5′CpG岛甲基化的情况;(3)Western blot方法检测细胞ERα蛋白的表达;(4)MTT法检测MDA-MB-231乳腺癌细胞经TCS和5-Aza-C处理后,雌激素拮抗剂TAM对肿瘤细胞生长的影响。
结果:(1)RT-PCR法显示ERα阴性人乳腺癌细胞株MDA-MB-231ERαmRNA无表达,ERα阳性人乳腺癌细胞株MCF-7 ERαmRNA表达呈强阳性,5-Aza-C和TCS处理后的MDA-MB-231细胞ERαmRNA恢复表达;(2)MSP鉴定ERα甲基化状态:MDA-MB-231细胞仅甲基化引物扩增出特异PCR条带;MCF-7细胞仅非甲基化引物扩增出特异PCR条带;5-Aza-C和TCS处理后的MDA-MB-231细胞仅非甲基化引物扩增出特异PCR条带;(3)Western-blot方法分析显示MDA-MB-23细胞ERα蛋白不表达;MCF-7细胞ERα蛋白表达呈强阳性;5-Aza-C和TCS作用MDA-MB-231细胞后ERα蛋白恢复了表达;(4)MTT检测TCS和5-Aza-C处理ERα阴性MDA-MB-231细胞后,TAM恢复了对肿瘤细胞的生长抑制作用,且TCS处理组抑制作用比5-Aza-C组强(p＜0.05)。
结论:乳腺癌ERα基因表达缺失与ERα基因高甲基化有关,TCS可逆转ERα的甲基化,使ERαmRNA和蛋白重新表达,并可恢复对内分泌治疗药物TAM的敏感性。
第三章TCS对ERα阴性乳腺癌裸鼠移植瘤生长及ERα表达的影响
目的:研究TCS对ERα阴性乳腺癌裸鼠移植瘤生长及移植瘤内ERα基因表达和CpG岛甲基化的影响。
方法:(1)将MDA-MB-231乳腺癌细胞悬液接种于20只BALB/C(nu/nu)雌性裸鼠(SPF级)。18只裸鼠成瘤后将其分为三组:对照组、TCS组和盐水组,每组6只。分别于移植瘤旁注射TCS(0.4mg/kg)、盐水0.2ml,隔日注射一次,共7次;(2)检测裸鼠移植瘤体积和瘤重,计算肿瘤生长抑制率,检测各组裸鼠肝肾功能和血常规;(3)RT-PCR检测TCS对裸鼠乳腺癌移植瘤ERαmRNA表达的影响;(4)MSP检测TCS对裸鼠乳腺癌移植瘤ERα甲基化状态的影响;(5)Western blot检测TCS对裸鼠乳腺癌移植瘤组织ERα蛋白表达的影响。
结果:(1)细胞接种裸鼠成瘤成功率为90%(18/20);(2)TCS组裸鼠移植瘤体积和瘤重较对照组明显缩小(P＜0.05);(3)TCS对荷瘤裸鼠肾功能、血液系统无明显影响(P＞0.05),可引起谷丙转氨酶轻度升高(P＜0.05);(4)RT-PCR结果显示TCS治疗组裸鼠乳腺癌移植瘤ERαmRNA重新表达,而对照组和盐水组则无ERαmRNA表达;(5)裸鼠移植瘤模型的对照组和盐水组仅甲基化引物扩增出特异PCR条带,而TCS组仅非甲基化引物扩增出特异PCR条带;(6)治疗组乳腺癌组织可见ERα蛋白表达,对照组和盐水组ERα蛋白表达阴性。
结论:TCS可抑制乳腺癌肿瘤生长且在体内应用有一定的安全性;体内实验进一步证实TCS可以逆转ERα基因的高甲基化,使ERαmRNA和蛋白重新表达。
Breast cancer is the most common cancer in women,of which the morbidity increases year by year.Many researches presented estrogen is important in breast cancer.Two thirds of primary breast cancer have the expression of estrogen receptor(ER) and the growth of tumor cell is hormonal dependent.The function of estrogen depends on binding with its receptor and promoting the progression of breast tumor.The clinical therapy of breast cancer patients(ER positive) usually includes endocrinotherapy,which is equal to chemotherapy.However,one third breast cancer patients(ER negative) are not sensitive to endocrine therapy.
Estrogen receptor alpha(ERα) gene is located on the chromosome 6q24-27.The proteinum expressed by ERαgene is usually an important molecule marker to evaluate hormonal dependence and predicts the effect of endocrine therapy on breast cancer.Current researches report the mechanism of ERαgene expression silencing is not due to gene mutation, but probably associate with the genetic transcription.Because of the 5'CpG island methylation in the core promotor of ERαgene,the genetic transcription is suppressed which induce to the silence of ERαgene.
Trichosanthin(TCS) is a kind of traditional Chinese medicine,which we choose as experimental drug of demethylation.The purpose is to investigate the the effect of TCS on proliferation of MDA-MB-231 and MCF-7 breast cancer cell,the demethylating effect on ERαgene in MDA-MB-231 breast cancer cell,the relationship between demethylation and gene expression;meanwhile,to explore a new therapeutic strategy for breast cancer by demethylating treatment.The design of the study include:
(1) Investigate the effect of TCS acting on breast cancer cell MDA-MB-231 and MCF-7;
(2) Measure the demethylation of ERαmRNA and protein.Analyze the relationship between ERαmRNA expression and methylation;
(3) Explore the demethylating effect of TCS on MDA-MB-231 breast cancer cell and the sensitivity to endocrine therapy;
(4) Study the suppressive effect of TCS on transplanted human breast cancer in nude mice and the security in vivo.
The PartⅠ
Objective:Investigate the effects of TCS on proliferation,apoptosis of MDA-MB-231 and MCF-7 breast cancer cell.
Methods:(1) Human breast cancer cell lines MDA-MB-231 and MCF-7 were growing at 37℃and 5%CO_2.The morphological change of cells were observed by optical microscope after treated with TCS(90μg/ml) for 72 hours;(2) MTT was performed for the proliferation of MDA-MB-231 and MCF-7 cell after treated with TCS;(3) The period and apoptosis rate of MDA-MB-231 and MCF-7 cell were tested by Flow Cytometry method(FCM) after treated with TCS(90μg/ml) for 48 hours.
Results:(1) The cellular morphology changed after treated with TCS.Denaturation and necrosis occurred in some cells;(2) TCS could inhibit the growth of MDA-MB-231 and MCF-7 cell by MTT test;(3) TCS could suspend MDA-MB-231 and MCF-7 cell generation at G0/G1 period.It also induced MDA-MB-231 and MCF-7 cell apoptosis by FCM test.
Conclusion:TCS could inhibit MDA-MB-231 and MCF-7 cell proliferation in a time- and concentration-dependent manner.TCS could induce MDA-MB -231 and MCF-7 cell apoptosis and suspend the cell generation at G0/G1 period.
The PartⅡ
Objective:Explore the relationship between ERαgene expression silencing and hypermethylation.Study the effect of TCS on ERαgene expression and 5'CpG island methylation in MDA-MB-231 breast cancer cell.Test the possibility that TCS may induce MDA-MB-231 breast cancer cell sensitive to endocrine therapy.
Methods:We chose MDA-MB-231 breast cancer cell(ERαnegative) and MCF-7 breast cancer cell(ERαpositive).MDA-MB-231 cell was treated with TCS(90μg/ml) or 5-Aza-C(5μM).(1) The ERαmRNA level of the cells was measured by RT-PCR;(2) The 5'CpG island methylation of ERαgene was tested by Methylation Specific PCR(MSP) test;(3) Western blot method was performed for ERαprotein in the cells;(4) MTT method tested whether Tamoxifen could induce the growth inhibition of MDA-MB-231 cell after treated with TCS or 5-Aza-C.
Results:(1) The expression of ERαmRNA in MDA-MB-231 cell was silencing by RT-PCR test,while the expression of ERαmRNA in MCF-7 cell was positive.The re-expression of ERαmRNA in MDA-MB-231 cell treated with TCS or 5-Aza-C was shown by RT-PCR; (2) Methylation of ERαgene was found in MDA-MB-231 cell with MSP technique.Demethylation of ERαgene was found in MCF-7 cell and MDA-MB-231 cell treated with TCS or 5-Aza-C;(3) The expression of ERαprotein was negative in MDA-MB-231 cell,but positive in MCF-7 cell determined by western blot method,and re-expressed when treated with TCS or 5-Aza-C;(4) Tamoxifen could inhibit the proliferation of MDA-MB-231 cell treated with TCS efficiently through MTT method and the suppressive effect of TCS was stronger than 5-Aza-C(p＜0.05).
Conclusion:ERαgene expression silencing was related to the hypermethylation of ERαgene.TCS could induce the re-expression of ERαmRNA and protein in MDA-MB-231 cell by demethylating ERαgene.TCS could induce MDA-MB-231 breast cancer cell sensitive to Tamoxifen.
The PartⅢ
Objective:Study the suppressive effect of TCS on transplanted human breast cancer in nude mice and effect on the expression of ERα.
Methods:(1) 20 nude mice were transplanted with human breast cancer MDA-MB-231 cell to construct tumor models.Once the tumor model in nude mice was established,18 nude mice were devided into 3 groups:control group(without treatment),TCS group and saline group. TCS(0.4mg/kg) and sodium chloride were injected around the tumor.A total of 7 injections were given,once every 2 days;(2) Observations were made on tumor suppression rate,tumor volume and weight,liver and renal function,blood routine test;(3) The ERαmRNA level in transplanted neuplasma was measured by RT-PCR assays;(4) The methylation of ERαgene in transplanted neuplasma was performed by MSP test;(5) Immunohistochemistry and western blot were used to detemine the expression of ERαprotein in the 3 groups.
Results:(1) The subcutaneous tumor model in nude mice was successfully established(18/20);(2) The tumor volume and weight in TCS group decreased(p＜0.05);(3) Renal function and blood system were normal(P＞0.05) in TCS group.SGPT in TCS group was a little higher than control group(P＜0.05);(4) The re-expression of ERαmRNA was shown by RT-PCR in TCS group;(5) Demethylation of ERαgene was found in transplanted neuplasma treated with TCS by MSP test;(6) The expression,of ERαprotein was found in transplanted neuplasma treated with TCS by immunohistochemistry and western blot test.
Conclusions:TCS could effectively suppress human breast cancer cells growth rate in subcutaneous tumor of nude mice and it was relatively safe to use it.TCS could induce the re-expression of ERαmRNA and protein in vivo by demethylating ERαgene.
引文
[1]熊用周,潘平,徐绍嫣,等.天花粉蛋白对体外合体滋养层细胞影响的研究.动物学报,1976,22:172-178
[2]Mosmann T.Rapid colorimetric assay for cellular growth and survival:application to proliferation and cytotoxicity assays.Journal of Immunological Methods,1983,65(1-2):55-63
[3]魏熙胤,牛瑞芳.流式细胞仪的发展历史及其原理和应用进展.现代仪器,2006,4,8-11
[4]Zhang XJ,Wang JH.Homology of trichosanthin and ricin A chain.Nature,1986,321:477-478
[5]Chow TP,Feldman RA,Lovett M,et al.Isolation and DNA sequence of a gene encoding alpha-trichosanthin,a type Ⅰribosome-inactivating protein.J Biol Chem,1990,265:8670-8674
[6]Shaw PC,Yung MH,Zhu RH,et al.Cloning of trichosanthin cDNA and its expression in Escherichia coli.Gene,1991,97:267-272
[7]郑洪刚,王斌,邵鹏柱,等.天花粉蛋白基因的克隆及序列分析.遗传学报,1994,21:42-51
[8]Collins EJ,R obertus JD,Lo Presti M,et al.Primary amino acid sequence of alpha-trichosanthin and molecular models for abrin A-chain and alpha-trichosanthin.J Biol Chem,1990,265:8665-8669
[9]Shaw PC,Chan W L,Yeung H W,et al.Minireview:trichosanthin,a protein with multiple pharmacological properties.Life Sci,1994,55(4):253
[10]Tsao SW,Yan KT,Yeung HW.Selective killing of chodocarcinoma cells in vitro by tdchosanthin,a plant protein purified from root tubers of the Chinese medicinal herb Trichosanthes kidlowii.Toxicon,1986,24:831-840
[11]Zheng YT,Zhang WF,Ben KL,et al.In vitro immunotoxicity and cytotoxicity of trichosanthin against human normal immunocytes and leukemia-lymphoma cells.Immunopharmacology and Immunotoxicology,1995 17:69-79
[12]Chan WL,Shaw PC,Tam SC,et al.Trichosanthin interacts with and enters cells via LDL receptor family members.Biochemical and Biophysical Research Communications,2000,270:453-457
[13]Ru QH,Luo GA,Liao JJ,et al.Capillary electrophoretic determination of apoptosis of HeLa cells induced by trichosanthin.Journal of Chromatography.2000,894:165-170
[14]Dou CM,Li JC.Effect of extracts of trichosanthes root tubers on HepAH cells and HeLa cells.World Journal of Gastroenterology.2004,10:2091-2094
[15]McGrath MS,Hwang KM,Caldwell SE,et al.GLQ223:an inhibitor of human immunodeficiency virus replication in acutely and chronically infected cells of lymphocyte and mononuclear phagocyte lineage.Proc Natl Acad Sci USA,1989,86:2844-2848
[16]李雪梅,黄利鸣.天花粉蛋白的研究进展.河北医药,2006,28(4):322-323
[17]Chan WL,Zheng YT,Huang H,et al.Relationship between trichosanthin cytotoxicity and its intracellular concentration.Toxicology,2002,177:245-251
[18]胡梅洁,张曙,章永平,等.天花粉蛋白和重组干扰素α-2b联合应用对消化道癌细胞的协同杀伤作用.肿瘤,1997,17(6):465-467
[19]涂水平,江石湖,乔敏敏,等.天花粉蛋白对胃癌多药耐药细胞的细胞毒和诱导凋亡作用.世界华人消化杂志,2000,8(2):150-152
[20]Bender CM,Pao MM,Jones FA.Inhibition of DNA methylation 5-Aza-2'-deoxycytidine suppresses the growth of human tumor cell lines.Cancer Res,1998,58(1):95-101
[21]夏晓峰,王少雄,等.天花粉蛋白在酸性条件下可自发插入脂单层.科学通报,1999,44(16):1740-1743
[22]戴荣禧,徐国江,刘莲英,等.天花粉蛋白对滋养层细胞转移损伤机制的研究.实验生物学报,1993,26:411-416
[23]许良中.实用肿瘤病理方法学(第一版).上海:上海医科大学出版社,1997.578-595
[24]Darzynkiewicz Z,Juan G,Li X,et al.Cytometryin cell necrobiology:analysis of apoptosis and accidental cell death(Necrosis).Cytometry,1997,27(1):1-20
[25]ZHANG SHU,HU MEIJIE,WU YUXIN,et al.Chinese Journal of Digestive Diseases,2001,2:125-128
[26]张曙,胡梅洁.天花粉蛋白诱导的胃癌细胞凋亡与bcl-2表达下降有关.中华消化杂志,2000,20(6):380
[27]Hu MJ,Zhang S,Wu YX,Jiang SH.Synergic cytotoxicity to gastric cancer cells by combined use of trichosanthin and recombinant interferon alpha-2b.J Shanghai Second Med University,2000,12:44-47
[28]Luo WD,Ren.CM,Zhu M,et al.Study on NB4 cell apoptosis induced by trichosanthin.Zhongguo Shi Yan Xue Ye Xue Za Zhi,2005,13:278-281
[29]孔梅,柯一保,周关云,等.天花粉蛋自诱发白血病细胞K562凋亡的研究.实验生物学报,1998,31:233-243
[30]黄益玲,黄利鸣,石新兰,等.天花粉蛋白对人宫颈癌HeLa细胞增殖和细胞凋亡的影响.中国药理学通报,2005,21:253-254
[31]毕黎琦,李洪军,张玉华,等.中药天花粉蛋白对黑色素瘤细胞凋亡及细胞周期的影响.中国中西医结合杂志,1998;18:35-37
[32]李振红,陆阳,刘晶星.天花粉化学成分与药理活性.国外医药(植物药分册),2003,18(1):1-4
[33]Li MX,Yeung HW,Pan LP,et al.Trichosanthin,a potent HIV-Ⅰinhibitor can cleave supercoiled DNA in vitro.Nucleic Acids Res,1991,19(22):6309-6312
[34]何肾辉,曾耀英,孙荭,等.天花粉蛋白诱导人类白血病细胞株HL-60细胞凋亡的研究.中国病理生理杂志,2001,17(3):200
[35]魏明卉,李英,刘征.核糖体失活蛋白及研究进展.南都学坛(自然科学版),2001,21(6):64-67
[36]胡红雨.鲁子贤.1H NMR法研究天花耢蛋白催化5'-AMP糖苷酶的裂解.科学通报,1994,39:1720-1722
[37]Meguire WL.Hormone receptors:their role in predicting prognosis and response to endocrine therapy.Semin Oncol,1978,5:428-433
[38]Sommer S,Fuqua SA.Estrogen receptor and breast cancer.Semin Cancer Biol,2001,11:339-352
[39]Lapidus RG,Nass SJ,Davidson NE.The loss of estrogen and progesterone receptor gene expression in human breast cancer.J Mamm Gland Biol Neoplasia,1998,3:85-94
[40]Ottaviano YL,Issa JP,Pad FF,et al.Methylation of the estrogen receptor gene CpG island marks loss of estrogen receptor expression in human breast cancer cells.Cancer Res,1994,54(10):2552-2555
[41]Ferguson AT,Lapidus RG,Baylin SB,et al.Demethylation of the estrogen receptor gene in estrogen receptor-negative breast cancer cells can reactivate estrogen receptor gene expression.Cancer Research,1995,55(11):2279-2283
[42]韩明强,黄韬,田元.雌激素水平对三苯氧胺抑制乳腺癌细胞的体外研究.中国普外基础与临床杂志,2005,12(6):589-590,594
[43]Matins-Guiu X,Catasus L,Bussaglia E,et al.Molecular pathology of endometrial hyperplasia and carcinoma.Hum Pathol,2001,32(6):569-577
[44]Becker K,Kellerq,Hoefler H.The use molecular biology in diagnosis and prognosis of gastric cancer.Surg Oncol,2000,9(1):5-11
[45]Nagane Y,Utsugisawa K,Tobgi H.PCR amplification in bisulfite methylcytosine mapping in the GC-rich promoter region of a myloid precursor protein gene in autopsy human brain.Brain Res Prow,2000,5(2):167-171
[46]Chide Y,Ishizaki M,Nakazawa M.Expression and methylation of the beta-subunit gene of prolyl 4-hydroxylase:in erythrocytes,tendon and cornea of chickembryos.Connect Tissue Res,1992,28(3):191-204
[47]Asch BB,Barcellos-Hoff MH.Epigenetics and breast cancer.J Mammary Gland Biol Neoplasia,2001,6:151-152
[48]Cameron EE,Bachman KE,Myohanen S,et al.Synergy of demethylation and histone deacetylase inhibition in the re-expression of genes silenced in cancer.Nat Genet,1999,21:103-107
[49]Donaghue C,Westley BR,May FE.Selective promoter usage of the human estrogen receptor-alpha gene and its regulation by estrogen.Mol Endocrinol,1999,13:1934-1950
[50]Osborne C K.Steroid hormone receptors in breast cancer management.Breast Cancer Research and Treatment,1998,51(3):227-238
[51]Ali S,Coombes RC.Estrogen receptor alpha in human breast cancer:occurrence and significance.J Mammary Gland Biol Neoplasia,2000,5(3):271-281
[52]Hayashi SI,Eguchi H,Tanimoto K,et al.The expression and function of estrogen receptor alpha and beta in human breast cancer and its clinical application.Endocr Relat Cancer,2003,10(2):193-202
[53]Roodi N Bailey LR,Kao WY,et al.Estrogen receptor gene analysis in estrogen receptor-positive and receptor-negative primary breast cancer.J Natl Cancer Inst,1995,87(6):446-451
[54] Watts CK, Handel ML, King RJ, et al. Oestrogen receptor gene structure and function in breast cancer. J Steroid Bio chem Mol Biol, 1992,41(3-8): 529-536
[55] Iwase H, Kobayashi S, Iwata H, et al. Molecular analysis of the estrogen receptor(ER) gene in association with ER negativity in breast cancer. Gan To Kagaku Ryoho. 1996,23(Suppl 1): 61-65
[56] Dowsett M, Dafada A, Chan CM, et al. Oestrogen receptor mutants and variants in breast cancer. Eur J Cancer, 1997, 33(8): 1177- 1183
[57] Stoner M, Saville B,Wormke M, et al. Hypoxia induces proteasome dependent degradation of estrogen receptor alpha in ZR275 breast cancer cells. Mol Endocrinol, 2002,16:2231-2242
[58] Roger P, Daures JP, Maudelonde T, et al. Dissociated over expression of cathepsin D and estrogen receptor alpha in preinvasive mammary tumors. Hum Pathol, 2000,31:593-600
[59] Hewitt SC, Bocchinfuso WP, Zhai J, et al. Lack of ductal development in the absence of functional estrogen receptor alpha delays mammary tumor formation induced by transgenic expression of ErbB2/neu. Cancer Res, 2002, 62(10): 2798-2805
[60] Lapidus RG» Nass SJ, Butash KA. et al. Mapping of ER gene CpG island methylation-specific polymerase chain reaction. Cancer Res. 1998, 58(12): 2515- 2519
[61] Xiong Z, Laird PW. COBRA: a sensitive and quantitative DNA methylation assay-Nucleic Acids Res, 1997,25(12): 2532-2534
[62] Suzuki H, Itoh F, Toyota M, et al. Quantitative DNA methylation analysis by fluorescent polymerase chain reaction single-strand conformation polymorphism using an automated DNAs equence. Electrophoresis, 2000,21(5): 904-908
[63] Gonzalgo ML, Jones PA. Rapid quantitation of methylation differences at specific sites using methylation-sensitive single nucleotide primer extention(Ms-SnuPE). Nucleic Acids Res, 1997,25(12): 2529-2531
[64] Bianco T, Hussey D, Dobrovic A. Methylation-sensitive single strand conformation analysis(MS-SSCA): A rapid method to screen for and analyze methylation. Hum Mutat, 1999,14(4): 289-293
[65] Eads CA, Danenberg KD, Kawakami K, et al. MethyLight: a high-throughput assay to measure DNA methylation. Nucleic Acids Res, 2000,28(8): E32
[66] Esteller M, Catasus L, Matias-Guiu X, et al.hMLHl promoter hypermethylation is an early event in human endometrial tumorigenesis. Am J Pathol, 1999,155:1767-1772
[67] Jones PA, Laird PW. Cancer epigenetics comes of age. Nat Genet, 1999, 21(2): 163-167
[68] Herman JG, Graff JR, Myohanen S, et al. Methylation-specific PCR: a novel PCR assay for methylation status of CpG islands. Proc Natl Acad Sci USA, 1996, 93(18): 9821-9826
[69] Singal R, Ginder GD. DNA methylation. Blood, 1999, 93:4059-4070
[70] Hortobagyi GN. Treatment of breast cancer. N Engl J Med, 1998, 339: 974- 984
[71] Weigel RJ, deConinck EC. Transcriptional control of estrogen receptor in estrogen receptor-negative breast carcinoma. Cancer Res 1993,53: 3472-3474
[72] Wolffe AP,Matzke MA. Epigenetics: regulation through repression. Science, 1999,286(5439): 481-486
[73] Zhao C, Lam EW , SuntersA. et al. Expression of estrogen recptor beta isoformsin normal breast epithelial cells and breast cancer : regulation by methylation. Oncogene, 2003, 22(48): 760007606
[74] Jones PA, Laird PW. Cancer epigenetics comes of age. Nat Genet, 1999, 21: 163-167
[75] Baylin SB, Herman JG. DNA hypermethylation in tumorigenesis: epigenetics joins genetics. Trend Genet, 2000,16: 1680174
[76] Momparler RL, Bovenzi V. DNA methylation and cancer. J Cell Physiol, 2000, 183(2): 45-54
[77] Bird AP CPC-rich islands and the function of DNA methylation. Nature, 1986, 321 (6067) : 206-213
[78] Dominguez G, Silva J, Garcia JM, et al. Prevalence of aberrant methylation of pl4ARF over p16INK4a in some human primary tumors. Mutat Res Fundam Mol Mech Mutagen, 2003,530 (1) : 9-17
[79] Nicolai M, Marco B, Frank R, et al. Hypermethylation and histone deacetylation lead to silencing of the maspin gene in human breast cancer. Biochemical and Biophysical Research Communications, 2002, 297(1): 125-128
[80]Christman JK.5-Azacytidine and 5-aza-2'-deoxycytidine as inhibitor of DNA methylation:mechanistic studies and their implications for cancer therapy.Oncogene,2002,21:5483
[81]温国林,陈晓光.DNA甲基化及其抑制剂研究进展.国外医学遗传学分册,2001,24(4):180-184
[82]Rochefort H,Glondu M,Sahla ME,et al.How to target estrogen receptor-negative breast cancer? Endocr Relat Cancer,2003,10(2):261-266
[83]Varga Z,Zhao J,Ohlschlegel C,et al.Preferential HER-2/neu overexpression and/or amplification in aggressive histological subtypes of invasive breast cancer.Histopathology,2004,44(4):332-338
[84]Laurent MV,Lucas A,Huttler S,et al.Procathepsin D interacts with prosapostn in can c cells but its internalization is not mediated by LDL receptor- related protein.Exp Cell Res,2002,277(2):210-219
[85]Shibata A,Nagaya T,Imai T,et al.Inhibition of NF-kappaB activity decreases the VEGF mRNA expression in MDA-MB-231 breast cancer cells.Breast Cancer Res Treat,2002,73(3):237-43
[86]Hirvonen R,Talvensaari MA,Paakko P,et al.Matrix metalloproteinase-2(MMP-2) in T_(1-2)N_0 breast carcinoma.Berast Cancer Res Treat,2003,77(1):85-91
[87]Franke H R,Kole S,Ciftci Z,et al.In vitro effects of estradiol,dydrogesterone,tamoxifen and cyclophosphamide on proliferation vs.death in human breast cancer cells.Cancer Letters,2003,190(1):113-118
[88]Shao Z,Jiang M,Yu L,et al.Estrogen receptor-negative breast cancer cells transfected with estrogen receptor exhibit decreased tumour progression and sensitivity to growth inhibition by estrogen.Chin Med Sci J,1997,12(1):11
[89]Tsao SW,Ng TB,Yeung HW.Toxicities of trichosanthin and alpha-momorcharin,abortifacient proteins from Chinese medicinal plants,on cultured tumor cell lines.Toxicon,1990,28:1183-1192
[90]汪钊猷主编.天花粉蛋白.上海:科学出版社,1990.4-7
[91]孙健,吴志全,李雁,等.天花粉蛋白与重组人干扰素α-1b协同抑制人肝细胞癌的实验研究.中国临床医学,2003,10(3):278-280
[92]徐振武,吴裕忻,陈陵际,等.天花粉蛋白对荷人结肠癌SW-1116裸鼠的体内实验研究肿瘤学杂志,1997,3(2):110-111
[93]Ln JB,LiMY,et al.Detenmination of the binding epitope for anti-trichosanthin monochonal antibodyT(8)C(12).Sheng Wu Hua Xue Yu Sheng Wu Wu Li Xue Bao,1995,30:137-141
[94]唐文皓,吴志全,等.樊嘉天花粉蛋白对荷人肝癌裸小鼠的抑癌作用.中华肝胆外科杂志,2003,9(4):228-230
[95]Ke YB,Chen J K,Hie HL,et al.Structure-function relationship of trichosanthin.Life Sci,1997,60:465-472
[96]张劲松,刘望夷.天花粉蛋白在真核细胞核糖体28S rRNA上的作用位点.生物化学与生物物理进展,1992,19:131
[97]田维毅,马春玲,白惠卿.天花粉及其组份对小鼠NK细胞杀伤活性的影响.贵州医学杂志,2001,25:982-984
[98]田维毅,马春玲,白惠卿.天花粉及其组份对小鼠NK细胞杀伤活性的影响.贵州中医,2001,25(11):289
[99]周广宇,汪丽,毕黎琦,等.天花粉蛋白对人PBMC分泌细胞因子水平及离子通道的影响.中国免疫学杂志,2001,17:650-652
[100]郭峰,刘小平,徐瑛.天花粉对小鼠艾氏腹水癌的疗效和对红细胞免疫功能的影响.中西医结合杂志,1989,9:418-420
[101]骆和生,王建华,邓文龙,等.中药方剂的药理与临床研究进展.广州:华南理工大学出版社,1991.78
[102]Myer RA,Sergios PA,Coonan K,et al.Trichosanthin treatment of HIV-induced immunedys regulation.Eur J Invest 1992,22(2):113
[103]奚正德,马宝骊,曹鹤年,等.天花粉蛋白羟胺裂解片段制备及其对补体的激活作用.中国免疫学杂志,1997,13(1):47
[104]郭峰,刘小平,徐瑛.天花粉对小鼠艾氏腹水癌的疗效和对红细胞免疫功能的影响.中西医结合杂志,1989,9(7):418-420
[105]徐瑛,郭峰,叶天星。天花粉治疗小鼠艾氏腹水癌对红细胞增强吞噬作用的研究.肿瘤,1990,10(5):224
[106]邬伟秀,郭峰.天花粉对血液肿瘤患者红细胞免疫功能影响的实验研究.上海免疫学杂志,1995,5(3):156
[107]洪建,富赛里,陆佩华,等.天花粉蛋白对T细胞活化的抑制作用与信号传导.中华微生物学和免疫学杂志,1998,18(2):111
[108]胡梅洁,张曙.天花粉蛋白与重组干扰素α-2b联用对荷人胃腺癌SGC-7901 裸小鼠的抑癌作用.Shanghai Med J,1998,21(8):456-457
[109]Jones PA,Taylor SM,Mohandas T,et al.Cell cycle-specific reactivation of an inactive X-chromosome locus by 5-aza-deoxycytidine.Proc Natl Acad Sci USA,1982,79:1215-1219
[110]Yoshiura K,Kanai Y,Ochiari A,et al.Silencing of the E-cadherin invasion-suppressor gene by CpG methylation in human carcinomas.Proc Natl Acad Sci USA,1995,92:7416-7419
[111]Au WY,Fung A,Man C,et al.Aberrant p15 gene promoter methylation in therapy-related myelodyspiastic syndrome and acute myeloid leukaemia:clinicopathological and karyotypic associations.Br J Haematoi,2003,120(6):1062-1065
[112]Lemaire M,Momparler LF,Farinha NJ,et al.Enhancement of antineoplastic action of 5-aza-2'-deoxycytidine by phenylbutyrate on L1210 leukemic cells.Leuk Lymphoma,2004,45(I):147-154
[113]Miyamoto K,Ushijima T.DNA methylation and cancer-DNA methylation as a target of cancer chemotherapy.Gan To Kagaku Ryoho,2003,30(13):2021-2029
[114]Silverman LR,Holland JF,Davis RB,et al.Azacytidine induces trilineage responses in patients with myelodysplastic syndromes:the Cancer and Leukemia Group B experience.Hematologica,1991,76:42
[115]Jeffrey G,Zella RZ,Richard KS,et al.Treatment of myelodysplastic syndromes with 5-azacytidine.Leukemia Research,2002,26:893-897
[116]Tamura T,Kataoka A,Shu LY.et al.An in vitro screening method for DNA cytosine-C5-methylase inhibitor.Nat Prod Lett,2002,16(1):25
[117]Jeong LS,Yoo S J,Lee KM,et al.Design,synthesis,and biological evaluation of fluoroneplanocin A as the novel mechanism-based inhibitor of S-adenosylhomocysteine hydrolase.J med Ctiem,2003,46(2):201-203
[118]黄利鸣,邢学森,黄益玲.p16基因甲基化与Hela细胞凋亡相关性的研究.中国肿瘤临床,2005,32(13):721-724
[119]RX Dai,GL Xu,LY Liu,et al.Study on injury-mechanism of trichosanthin on trophoblast cells and choriocarcinoma cells in culture.Shiyan Sshengwu Xuebao,1993,26(4):411-427
[120]胡梅洁,吴云林,张曙,等.消化病与内镜(第一版).上海:上海科技文 献出版社.1998.281-284
[121]马宝骊,陈汐,杨黎明.天花粉蛋白类过敏反应分子机理的研究.医学研究通讯,1993,22(5):25
[122]黄健,金毓翠.天花粉蛋白宫颈注射引产使用地塞米松减少副反应的分析.现代妇产科进展,1994,3:227-228
[123]Byers VS,Levin AS,Malvino A,et al.A phase Ⅱstudy ofeffect of addition of trichosanthinto zidovudine in patients with HIV disease and failing antiretroviral agents.AIDS Res Hum Retroviruses,1994,10(4):413-420
[124]张淑景,金毓翠,于金德,等.天花粉引产对孕妇心、肝、肾功能影响的进一步研究.生殖与避孕,1994.14:19-24
[125]张素胤,张家骝.8种抗癌药物对裸小鼠肾包膜下接种的人肺腺癌移植瘤(LAX-83)生长的影响.中国药理学报,1989,15(5):450-453
[126]王茂钦,李莉,王洁,等.天花粉蛋白致豚鼠全身过敏反应的实验研究.安徽中医学院学报,1999,18:47-49
[127]徐振武,吴裕忻,陈陵际,等.天花粉蛋白抗结肠癌体内实验研究.海峡药学,1997,9:12-13
[128]时伟红,宋今丹.含天花粉蛋白的抗人大肠癌免疫毒素制备及其细胞毒性研究.中国医科大学学报,1996,25(2):111-113
[1]Wolffe A P,Matzke M A.Epigenetics:regulation through repression.Science,1999,286(5439):.481-486
[2]王树人.基因的表达与Epigenetics.四川生理科学杂志.2006,28(1):29-33
[3] J aenisch R ,Bird A. Epigenetic regulation of gene expression : how the genome integrates intrinsic and environmental signals. Not Rev Genet ,2003 ,33 :245-254
[4] Matias K P. Study of epigenetic changes leads to treatment advances in adult leukemias. Oncolog ,2003 ,48(1): 1-4
[5] Okano M,Bell ,DW,Haber DA ,Li E. DNA methyltransferases Dnmt3a and Dnmt3b are essential for de novo methylation and mammalian development. Cell ,1999,99(3):247-257
[6] Reik W, Dean W, Walter J. Epigenetic reprogramming in mammalian development. Science, 2001,293(5532): 1089-1093
[7] Rush LJ, Plass C. Restriction landmark genomic scanning for DNA methylation in cancer :past ,present ,and future applications. Anal Biochem,2002 ,307(2): 191 -201
[8] Robertson KD Jones PA. DNA methylation : past ,present and future directions. Carcinogenesis ,2000,21 (3) :461-467
[9] Peter A J , Daiya T. The role of DNA methylation in mammalian epigenetics. Science, 2001,293(5532): 1068-1070
[10] Ichimura T , Watanabe S , Sakamoto Y, et al. Transcriptional repression and heterochromatin formation by MBD1 and MCAF/AM family proteins. J Biol Chem, 2005,280(14): 13928-13935
[11] Tamura G, Yin J , Wang S , et al. E-Cadherin gene promoter hypermethylation in primary human gastric carcinomas. J Natl Cancer Inst, 2000,92(7):569-573
[12] Manini DN ,Rodenhiser DI ,Ainsworth PJ , et al. CpG methylation within the 5'regulatory region of BRCAL ,genies tumor specific and includes a putative CREB binding site. Oncogene, 1998,16(9): 1161 - 1169
[13] Clark SJ ,Harrison J ,Molloy PL. Spl binding is inhibited by CpG methylation. Gene,1997,195 (1) :67-71
[14] Singal R, Ginder GD. DNA methylation. Blood, 1999,93(12) :4059-4070
[15] Nan X,Ng HH ,Johnson CA , et al . Transcriptional repression by the met hyl CpG binding protein MeCP2 involves a histonedeacetylase complex. Nature ,1998 ,393 (6683):386-389
[16] Ng HH , Zhang Y, Hendrich B, et al . MBD2 is a transcriptional repressor belonging to the MeCPl histone deacetylase complex. Nat Genet, 1999,23(1): 58-61
[17] Momparler RL ,Bovenzi V. DNA methylation and cancer. J Cell Physiol, 2000, 183(2): 145-154
[18] Jones PA, Laird PW. Cancer epigenetics comes of age. Nat Genet,1999,21(2): 163-167
[19] Hsieh CL. Evidence t hat protein binding specifies sites of DNA demethylation. Mol Cell Biol,1999,19(1) :46-56
[20] Wolffe AP ,Jones PL ,Wade PA. DNA demethylation. Proc Natl Acad Sci USA, 1999,96(11): 5894-5896
[21] Jost J P ,Siegmann M ,Sun L ,et al . Mechanisms of DNA demethylation in chicken embryos. Purification and properties of a 5-methylcytosine-DNA glycosylase.J Biol Chem,1995,270(17): 9734-9739
[22] Zhu B ,Zheng Y ,Hess D ,et al. 5-methylcytosine-DNA glycosylase activity is present in a cloned G/T mismatch DNA glycosylase associated with the chicken embryo DNA demethylation complex. Proc Natl Acad Sci USA,2000,97(10) :5135-5139
[23]朱卫国. DNA甲基化, 基因调控和癌症. 世界华人消化杂志, 2002, 10(6): 680-683
[24] Grady WM, Willis S , Guilford PJ, et al. Methylation of the CDH1 promoter as the second genetic hit in hereditary diffuse gastric cancer.Nat Genet, 2000,26(1): 16-17
[25] Jones PA , Takai D. The role of DNA methylation in mammalian epigenetics. Science, 2001,293(5532):1068- 1070
[26] Jones PA ,Baylin SB. The fundamental role of epigenetic events in cancer. Nat Rev Genet,2002,3(6):415-428
[27] Lee WH ,Morton RA ,Epstein J I ,Brooks JD ,Campbell PA ,Bova GS , et al . Cytidine methylation of regulatory sequences near the piclass glutathione S-transferase gene accompanies human prostatic carcinogenesis. Proc Natl Acad Sci USA, 1994,91(24): 11733-11737
[28] Leone G,Teofili L ,Voso MT, et al. DNA methylation and demethylating drugs in myelodysplastic syndromes and secondary leukemias. Haematologica, 2002, 87(12): 1324-1341
[29] Robertson KD. DNA methylation and human disease. Nat Rev Genet, 2005, 6(8):597-610
[30] Li QL , Ito K, Sakakura C , et al . Causal relationship between the loss of RUNX3 expression and gastric cancer. Cell, 2002 ,109(1): 113-124
[31]李燕妮,张维铭.促甲基化反应的研究进展.国际检验医学杂志,2006,27(6):527-528,531
[32]Butcher DT,Mancini-DiNardo DN,Archer TK,et al.DNA binding sites for putative methylation boundaries in the unmethylated region of the BRCA1 promoter.Int J Cancer,2004,111(5):669-678
[33]Etoh T,Kanai Y,Ushijima S,et al.Increased DNA methyltransferase 1(DNMTI) protein expression correlates significantly with poorer tumor differentiation and frequent DNA hypermethylation of multiple CpG islands in gastric cancers.Am J Pathol,2004,164(2):689-699
[34]Nagai M,Nakamura A,Makino R,et al.Expression of DNA(5-cytosin)-methyltransferases (DNMTs) in hepatocellular carcinomas.Hepatol Res,2003,26(3):186-191
[35]Paz M F,Fraga M F,Avila S,et al.A systematic profile of DNA methylation in human cancer cell lines.Cancer Res,2003,63(5):1114-1121
[36]Yang B,Guo M,Herman J G,et al.Aberrant promoter methylation profiles of tumor suppressor genes in heaptocellular carcinoma.Am J Pathol,2003,163(3):1101-1107
[37]Esteller M,Herman JG.Cancer as an epigenetic disease:DNA methylation and chromatin alterations in human tumours.J Pathol,2002,196(1):127
[38]Wong N,Lam W C,Lai P B,et al.Hypomethylation of chromosome 1heterochromatin DNA correlates with qarm copy gain in human heaptocellular carcinoma.Am J Pathol,2001,159(2):465-471
[39]Nakagawa T,Kanai Y,Ushijima S,et al.DNA hypomethylation on pericentromeric satellite regions significantly correlates with loss of heterozygosisty on chromosome 9 in urothelial heteozygosity on chromosome 9 in urothelial carcinomas.J Urol,2005,173(1):243-246
[40]Rusmintratip V,Riggs AD,Sowers LC.Examination of the DNA substrate selectivity of DNA cytosine methyltransferases using mass tagging.Nucleic Acids Res,2000,28(18):3594-3599
[41]Florl AR,Lower R,Schmitz Drager BJ,et al.DNA methylation and expression of LINE21 and HERV2Kprovirus sequences in urothelial and renal cell carcinomas.Br J Cancer,1999,80(9):1312-1321
[42]Feinberg AP.Imprinting of a genomic domain of 11p15 and loss of imprinting in cancer:an introduction.Cancer Res,1999,59(7Suppl):1743-1746
[43]Kokalj-Vokac N,Almeida A,Viegas-Pequignot E,et al.Specific induction of uncoiling and recombination by azacytidine in classical satellite-containing constitutive heterochromatin.Cytogenet Cell Genet,1993,63(1):11-15
[44]吕斌斌,彭剑雄.DNA甲基化与基因沉默及肿瘤.国际检验医学杂志,2006,27(1):52-54,57
[45]Tao L,Yang S,Xie M,et al.Hypomethylation and overexpression of c-jun and c-myc protooncogenes and increased DNA methyltransferase activity in dichloroacetic and trichloroacetic acid-promoted mouse liver tumors.Cancer Lett,2000,158(2):185-193
[46]Jang S J,Soria JC,Wang L,et al.Activation of melanoma antigen tumor antigens occurs early in lung carcinogenesis.Cancer Res,2001,61(21):7959-7963
[47]Feng Z,Hu W,Rom WN,et al.N-hydroxy-4-aminobiphenyl-DNA binding in human p53 gene:sequence preference and the effect of C5 cytosine methylation.Biochemistry,2002,41(20):6414-6421
[48]Florl AR,Steinhoff C,Muller M,et al.Coordinate hyper- methylation at specific genes in prostate carcinoma precedes L1NE-1 hypomethylation.Br J Cancer,2004,91(5):985-994
[49]Seiandrello G,Caradonna F,Mauro M,et al.Arsenic-induced DNA hypomethylation affects chromosomal instability in mammalian cells.Carcinogenesis,2004,25(3):413-417
[50]Ehrlich M.The ICF syndrome,a DNA methyltransferase 3B deficiency and immunodeficiency disease.Clin Immunol,2003,109(1):17-28
[51]Gaudet F,Hodgson JG,Eden A,et al.Induction of Tumors in Mice by Genomic Hypomethylation.Science,2003,300(5618):489-492
[52]Wu J,lssa J,Hermen J,et al.Expression of an exogenous eukaryotic DNA methyltransferase gene induces transformation of NIN3T3 ceils.Proc Natl Acad Sci USA,1993,90(19):8891-8895
[53]Oakeley EJ,Schmitt F,Jost JP.Quantification of 5-methylcytosine in DNA by the chloroacetaldehyde reaction.Biotechniques,1999,27(4):744-746
[54]Wainfan E,Dizik M,Stender M,et al.Methyl group in carcinogenesis:effects on DNA methylation and gene expression. Cancer Res, 1992,52(7Suppl): 2071s-2077s
[55] Richard L , Robey R , Sandor V , et al. Inhibitors of histone deacetylation , Depsipeptide (FR901228) , in the treatment of peripheral and cutaneous T-cell lymphoma: a case in report. Blood,2001,98(9):2865-2868
[56] Urnov FD , Wolffe AP. Chromatin remodeling and transcriptional activation: the cast (in order of appearance). Oncogene, 2001,20(24 Rev 3):2991-3006
[57] Li Q , Ahuja N , Burger PC , et all Methylation and silencing of the Thrombospondin-1 promoter in human cancer. Oncogene, 1999,18(21): 3284 -3289
[58] Pogribny I, Yi P , James SJ , et al. A sensitive new method for rapid detection of abnormal methylation patterns in global DNA and within CpG islands. Biochem Biophys Res Commun, 1999,262(3): 624-628
[59] Rein T , Natale DA , Gartner U , et al. Absence of an unusual"densely methlated island"at the hamster ori-beta. JBiol Chem, 1997,272(15): 10021 - 10029
[60] Yamaguchi Y., Wada T., Suzuki F., et al. Casein kinase II interacts with the bZIP domains of several transcription factors. Nucleic Acids Res, 1998, 26(16): 3862- 3864
[61] Herman JG, Graft JR , Myohanen S , et al. Methylation-specific PCR: a novel PCR assay for methylation status of CpG islands. Proc Natl Acad Sci USA, 1996 , 93(18):9821-9826
[62] Redner RL , Wang J , Liu JM. Chromatin remodeling and leukemia: new therapeutic paradigms. Blood, 1999,94(2): 417-428
[63] Rand K, Qu W, Ho T, et al. Conversion-specific detection of DNA methylation using real-time polymerase chain reaction (ConLight-MSP) to avoid false positives. Methods, 2002 ,27(2): 114-120
[64] Yan PS , Chen CM, Shi H , et al. Dissecting comples epigenetic alterations in breast cancer using CpG island microarrays. Cancer Res, 2001,61(23):8375-8380
[65] Gonzalgo ML , Jones PA1 Rapid quantitation of methylation differences at specific sites using methylation-sensitive single nucleotide primer extension (Ms- NuPE). Nucleic Acids Res, 1997,25(12): 2529-2531
[66] El-Maarri D , Herbiniaux U , Walter J , et al. A rapid quantitative non- adioactive bisuifite-SnuPE-IP RP HPLC assay for methylation analysis at specific CpG sites . Nucleic Acids Res, 2002 , 30(6):e25
[67]Grunstein M.Histone acetylation in chromatin structure and transcription.Nature,1997,389(6649):349-352
[68]Oefner Pi,Bonn GK,Huber CG,et al.Comparative study of capillary zone electrophoresis and high-performance liquid chromatoguaphy in the analysis of oligonucleotides and DNA.J Chromatogrl 1992,625(2):331-401
[69]邓大君,邓国仁,吕有勇,等.变性高效液相色谱法检测CpG岛胞嘧啶甲基化.中华医学杂志,2001,80(2):158-161
[70]Banmer A,Wiedemann U,HergersbergM,et al.A novel MSP-DHPLC method for the investigation of the methylation status of imprintedgenes enables the molecular detection of low cell mosaicisms.Hum Mutat,2001,17(5):423-430
[71]Slater H,Brtmo D,Ren H,et al.Improved testing for CMT1A and HNPP using multiplex ligation-dependent probe amplification(MLPA) with rapid DNA preparations:comparison withthe interphase FISH method.Hum Mutat,2004,24(2):164-171
[72]Slater HR,Bruno DL,Ren H,et al.Rapid,high throughput prenatal detection of aneuploidy using a novel quantitative method(ML PA).J Med Genet,2003,40(12):907-912
[73]Worsham MJ,Pals G,Schouten J P,et al.Delineating genetic pathways of disease progression in head and neck squamous cell carcinoma.Arch Otolaryngol Head Neck Surg,2003,129(7):702-708
[74]Schouten J P,Mcelgunn CJ,Waaijer R,et al.Relative quantification of 40nucleic acid sequences by multiplex ligation-dependent probe amplification.Nucleic Acids Res,2002,30(12):e57
[75]Nygren AO,Ameziane N,Duarte HM,et al.Methylation-Specific MLPA(MS-MLPA):simultaneous detection of CpG methylation and copy number changes of up to 40 sequences.Nucleic Acids Res,2005,33(14):e128
[1]Palmisano W A,Divine K K,Saccomanno G,et al.Predicting lung cancer by detecting aberrant promoter methylation in sputum[J].Cancer Res,2000,60(21):5954-5958
[2]Cavalli L R,Urban C A,Dai D,et al.Genetic and epigenetic alterations in sentinel lymph nodes metastatic lesions compared to their corresponding primary breast tumors[J].Cancer Genet Cytogenet,2003,146(1):33-40
[3]Primeau M,Gagnon J,Momparler R L.Synergistic antineoplastic action of DNA methylation inhibitor 5-AZA-2'-deoxycytidine and histone deacetylase inhibitor depsipeptide on human breast carcinoma cells[J].Int J Cancer,2003,103(2):177-184
[4]Yang X,Yan L,Davidson N E.DNA methylation in breast cancer[J].Endocr Relat Cancer,2001,8(2):115-127
[5]Herman J G,Baylin S B.Promoter region hypermethylation and gene silencing in human cancer[J].Curr Top Mierobiol Immunol,2000,24(9):35-54
[6]Woodcock D M,Linseumeyer M E,Doherty J P,et al.DNA methylation in the promoter region of the p16(CDKN2PMTS21PINK4A) gene in human breast tumours[J].Br J Cancer,1999,79(2):251-256
[7]王志伟,易静,陈玉英,等.乳腺癌中p16基因甲基化及其与临床病理学特征的关系[J].中华实验外科杂志,2002,19(1):81
[8] Takai D, Jones P A. Comprehensive analysis of CpG islands in human chromosomes 21 and 22[J]. Proc Natl Acad Sci USA, 2002,99(6): 3740-3745
[9] Jones P A, Laird P W. Cancer epigenetics comes of age[J]. Nat Genet, 1999 , 21(2): 163-167
[10] Yang X, Ferguson A T, Nass S J, et al. Transcriptional activation of estrogen receptor a in human breast cancer cells by histone deacetylase inhibition[J]. Cancer Res, 2000,60(24): 6890-6894
[11] Ferguson A T, Evron E, Umbricht C B, et al. High frequency of hypermethylation at the 14-3-3 o locus leads to gene silencing in breast cancer[J]. Proc Natl Acad Sci USA, 2000,97(11): 6049-6045
[12] Celis A, Rasmussen H H, Celis P, et al. Short-term culturing of low-grade superficial bladder transitional cell carcinomas leads to changes in the expression levels of several proteins involved in key cellular activities[J].Electrophoresis, 1999, 20(2): 355-361
[13] Ottaviano Y L, Issa J P, Parl F F, et al. Methylation of the estrogen receptor gene CpG island marks loss of estrogen receptor expression in human breast cancer cells[J]. Cancer Res, 1994, 54(10): 2552-2555
[14] Ferguson A T, Vertino P M, Spitzner J R, et al. Role of estrogen receptor gene demethylation and DNA methyltransferase- DNA adduct formation in 5-aza-2'-deoxycytidine-induced cytotoxicity in human breast cancer cells[J]. J Biol Chem, 1997,272(51): 32260-32266
[15] Onate S O, Tsai S Y, Tsai M J, et al. Sequence and characterization of a coactivator for the steroid hormone receptor superfamily[J]. Science, 1995, 270(5240): 1354-1356
[16] Du Y, Carling T, Fang W, et al. Hypermethylation in human cancers of the RIZ1 tumor suppressor gene, a member of a histone/protein methyltransferase superfamily [J]. Cancer Res, 2001, 61(22): 8094-8099
[17] Graff J R, Gabrielson E, Fujii H, et al. Methylation patterns of the E-cadherin 5' CpG island are unstable and reflect the dynamic, hetero- geneous loss of E-cadherin expression during metastatic progression[J]. J Biol Chem, 2000 ,275(4): 2727-2732
[18] Chen C L, Liu S S, Ip S M, et al. E-cadherin expression is silenced by DNA methylation in cervical cancer cell lines and tumours [J]. Eur J Cancer , 2003, 39(4): 517-523
[19]Nass S J,Herman J G,Gabrielson E,et al.Aberrant methylation of the estrogen receptor and E-cadherin 5'CpG islands increases with malignant progression in human breast cancer[J].Cancer Res,2000,60(16):4346-4348
[20]Toyooka K O,Toyooka S,Virmani A K,et al.Loss of expression and aberrant methylation of the CDH13(H-eadhedn) gene in breast and lung carcinomas[J].Cancer Res,2001,61(11):4556-4560
[21]Chen L M,Chai K X.Prostasin serine protease inhibits breast cancer invasiveness and is transcriptionally regulated by promoter dna methy- lation[J].Int J Cancer,2002,97(3):323-329
[22]Catteau A,Harris W H,Xu C F,et al.Methylation of the BRCA1 promoter region in sporadic breast and ovarian cancer;correlation with disease characteristics[J].Oncogene,1999,18(11):1957-1965
[23]Esteller M,Corn P G,Urena J M,et al.Inactivation of glutathione Stransferase P1 gene by promoter hypermethylation in human neoplasia[J].Cancer Res,1998,58(20):4515-4518
[24]李云青,张恩仲,肖翠英,等.乳腺癌患者BRCA1基因启动子区甲基化模式的初步研究[J].癌症,2001,20(10):1057-1060
[25]Magdinier F,Billard L M,Wittmann G,et al.Regional methylation of the 5'end CpG island of BRCA1 is associated with reduced gene expression in human somatic cells[J].FASEB J,2000,14(11):1585-1594
[26]Fujii H,Biel M A,Zhou W,et al.Methylation of the HIC-1 candidate tumor suppressor gene in human breast cancer[J].Oncogene,1998,16(16):2159-2164
[27]Esteller M,Sanchez-Cespedes M,Rosell R,et al.Detection of aberrant promoter hypermethylation of tumor suppressor genes in serum DNA from non-small cell lung cancer patients[J].Cancer Res,1999,59(1):67-70
[28]黄世杰.癌的表观遗传性治疗:过去,现在和将来[J].国外医学药学分册:2006,33(5):342-343
[29]Sommer S,Fuqua S A W.Estrogen receptor and breast cancer[J].Semin Cancer Biol,2001,11(5):339-352
[30]Stoner M,Saville B,Wormke M,et al.Hypoxia induces proteasome dependent degradation of estrogen receptor alpha in ZR275 breast cancer cells[J].Mol Endocrinol, 2002,16 (10) :2231-2242
[31] Roger P,Daures J P,Maudelonde T, et al. Dissociated overexp ression of cathep sin D and estrogen recep tor alpha in p reinvasive mammary tumors [J]. Hum Pathol, 2000, 31 (5) : 593-600
[32] Donaghue C, Westley B R, May F E. Selective p romoter usage of the human estrogen receptor-alpha gene and its regulation by estrogen[J]. Mol Endocrinol, 1999, 13 (11) :1934-1950
[33] Lapidus R G, Nass S J, Butash K A, et al. Mapping of ER gene CpG island methylation-specific polymerase chain reaction[J]. Cancer Res, 1998,58(12): 2515-2519
[34] Zhao C, Lam E W, Sunters A, et al . Expression of estrogen receptor beta isoforms in normal breast epithelial cells and breast cancer: regulation by methylation[J]. Oncogene, 2003,22(48): 7600-7606
[35] Nass S J, Ferguson A T, El Ashry D, et al. Expression of DNA methyltransferase (DMT) and the cell cycle in human breast cancer cells [J]. Oncogene, 1999,18(52): 7453-7461
[36] Yang X, Ferguson A T, Nass S J, et al. Transcriptional activation of estrogen receptor alpha in human breast cancer cells by histone deacety- lase inhibition[J]. Cancer Res, 2000,60(24):6890-6894
[37] Yang X, Phillips D L, Ferguson A T, et al. Synergistic activation of functional estrogen receptor (ER)-alpha by DNA methyltransferase and histone deacetylase inhibition in human ER-alpha-negative breast cancer cells[J]. Cancer Res, 2001, 61(19): 7025-7029
[38] Zhu W G, Lakshmanan R R, Beal M D, et al. DNA methyltransferase inhibition enhances apoptosis induced by histone deacetylase inhibitors [J]. Cancer Res, 2001, 61(4): 1327-1333
[39] Billard L M, Magdinier F, Lenoir G M, et al. MeCP2 and MBD2 expression during normal and pathological growth of the human mammary gland [J]. Oncogene, 2002,21(17): 2704-2712
[40] Macaluso M, Cinti C, Russo G, et al. pRb2/ p130-E2F4/ 5-HDAC1- SUV39H1-p300 and pRb2/ p130-E2F4/ 5-HDAC1-SUV39H1-DNMT1 multimolecular complexes mediate the transcription of estrogen receptor alpha in breast cancer[J]. Oncogene, 2003, 22(23): 3511-3517
[41]Castagnetta L A,Miceli M D,Sorci C M,et al.Growth of LNCaP human prostate cancer cells is stimulated by estradiol via its own receptor[J].Endocrinology,1995,136(5):2309-2319
[42]朱英坚,叶敏.雌激素受体、雄激素受体与前列腺癌[J].国外医学泌尿系统分册,2002,22(5):283-286
[43]Li LC,Chui R,Nakajima K,et al.Frequent methylation of estrogen receptor in prostate cancer:Correlation with tumor progression[J].Cancer Res,2000,60(3):702-706
[44]Belinsky S A,Nikula K J,Baylin S B,et al.Increased cytosine DNA-methyl-transferase activity is target-cell-specific and an early event in lung cancer[j].Proc Natl Acad Sci USA,1996,93(9):4045-4050
[45]Lau K M,LaSpina M,Long J,et al.Expression of estrogen receptor(ER)-αand ER-β in normal and malignant prostatic epithelial cells:Regulation by methylation and involvement in growth regulation[J].Cancer Res,2000,60(12):3175-3182
[46]Schiff R,Massarweh S,Shou J,et al.Breast cancer endocrine resistance:how growth factor signaling and estrogen receptor coregulators modulate response[J].Clin Cancer Res,2003,9(1pt2):447S-454S
[47]Shiozawa T,Itoh K,Horiuchi A,et al.Down-regulation of estrogen receptor by the methylation of the estrogen receptor gene in endometrial carcinoma[J].Anticancer Res,2002,22(1A):139-143
[48]Hori M,Iwasaki M,Shimazaki J,et al.Assessment of hypermethylated DNA in two promoter regions of the estrogen receptor alpha gene in human endometrial diseases[J].Gynecol Oncol,2000,76(1):89-96
[49]Sasaki M,Kotcherguina L,Dharia A,et al.Cytosine- phosphoguanine methylation of estrogen receptors in endometrial cancer[J].Cancer Res,2001,61(8):3262-3266
[50]Ghabreau L,Roux J P,Niveleau A,et al.Correlation between the DNA global methylation status and progesterone receptor expression in normal endometrium,endometrioid adenocarcinoma and precursors[J].Virchows Arch,2004,445(2):129-134
[51]Fiegl H,Gattringer C,Widschwendter A,et al.Methylated DNA collected by tampons-a new tool to detect endometrial cancer[J].Cancer Epidemiol Biomarkers Prev,2004,13(5):882-888
[52]Brueckner B,Lyko F.DNA methyltransferase inhibitors:old and new drugs for an epigenetic cancer therapy[J].Trends Pharmacol Sci,2004,25(11):551-554
[53]Issa J P,Garcia-Manero G,Giles F J.Phase 1 study of low-dose prolonged exposure schedules of the hypomethylating agent 5-aza-2'- deoxycytidine(decitabine) in hematopoietic malignancies[J].Blood,2004,103(5):1635-1640
[54]Grodstein F,Martinez M E,Platz E A.Postmenopausal hormone use and risk for colonrectal cancer[J].Annals Internal Medicine,1998,128(9):705-712
[55]Nanda K,Bastian L A,Hasselblad V,et al.Hormone replacement therapy and the risk of colonrectal cancer:a meta-analysis.Obstet Gynecol,1999,93(5 SUPPL):880-888
[56]Issa J P,Ottaviano Y L,Celano P,et al.Methylation of the estrogen receptor CpG island links aging and neoplasia in human colon[J].Nature Genet,1994,7(4):536-540
[57]苏琦,敖启林,贺修胜,等.胃癌ER基因表达与CpG岛甲基化的意义[J].中国肿瘤临床,2004,31(11):607-610
[58]于力.DNA甲基化与肿瘤[J].中华内科杂志,2005,44(6):403-404
[2]Mosmann T.Rapid colorimetric assay for cellular growth and survival:application to proliferation and cytotoxicity assays.Journal of Immunological Methods,1983,65(1-2):55-63
[3]魏熙胤,牛瑞芳.流式细胞仪的发展历史及其原理和应用进展.现代仪器,2006,4,8-11
[4]Zhang XJ,Wang JH.Homology of trichosanthin and ricin A chain.Nature,1986,321:477-478
[5]Chow TP,Feldman RA,Lovett M,et al.Isolation and DNA sequence of a gene encoding alpha-trichosanthin,a type Ⅰribosome-inactivating protein.J Biol Chem,1990,265:8670-8674
[6]Shaw PC,Yung MH,Zhu RH,et al.Cloning of trichosanthin cDNA and its expression in Escherichia coli.Gene,1991,97:267-272
[7]郑洪刚,王斌,邵鹏柱,等.天花粉蛋白基因的克隆及序列分析.遗传学报,1994,21:42-51
[8]Collins EJ,R obertus JD,Lo Presti M,et al.Primary amino acid sequence of alpha-trichosanthin and molecular models for abrin A-chain and alpha-trichosanthin.J Biol Chem,1990,265:8665-8669
[9]Shaw PC,Chan W L,Yeung H W,et al.Minireview:trichosanthin,a protein with multiple pharmacological properties.Life Sci,1994,55(4):253
[10]Tsao SW,Yan KT,Yeung HW.Selective killing of chodocarcinoma cells in vitro by tdchosanthin,a plant protein purified from root tubers of the Chinese medicinal herb Trichosanthes kidlowii.Toxicon,1986,24:831-840
[11]Zheng YT,Zhang WF,Ben KL,et al.In vitro immunotoxicity and cytotoxicity of trichosanthin against human normal immunocytes and leukemia-lymphoma cells.Immunopharmacology and Immunotoxicology,1995 17:69-79
[12]Chan WL,Shaw PC,Tam SC,et al.Trichosanthin interacts with and enters cells via LDL receptor family members.Biochemical and Biophysical Research Communications,2000,270:453-457
[13]Ru QH,Luo GA,Liao JJ,et al.Capillary electrophoretic determination of apoptosis of HeLa cells induced by trichosanthin.Journal of Chromatography.2000,894:165-170
[14]Dou CM,Li JC.Effect of extracts of trichosanthes root tubers on HepAH cells and HeLa cells.World Journal of Gastroenterology.2004,10:2091-2094
[15]McGrath MS,Hwang KM,Caldwell SE,et al.GLQ223:an inhibitor of human immunodeficiency virus replication in acutely and chronically infected cells of lymphocyte and mononuclear phagocyte lineage.Proc Natl Acad Sci USA,1989,86:2844-2848
[16]李雪梅,黄利鸣.天花粉蛋白的研究进展.河北医药,2006,28(4):322-323
[17]Chan WL,Zheng YT,Huang H,et al.Relationship between trichosanthin cytotoxicity and its intracellular concentration.Toxicology,2002,177:245-251
[18]胡梅洁,张曙,章永平,等.天花粉蛋白和重组干扰素α-2b联合应用对消化道癌细胞的协同杀伤作用.肿瘤,1997,17(6):465-467
[19]涂水平,江石湖,乔敏敏,等.天花粉蛋白对胃癌多药耐药细胞的细胞毒和诱导凋亡作用.世界华人消化杂志,2000,8(2):150-152
[20]Bender CM,Pao MM,Jones FA.Inhibition of DNA methylation 5-Aza-2'-deoxycytidine suppresses the growth of human tumor cell lines.Cancer Res,1998,58(1):95-101
[21]夏晓峰,王少雄,等.天花粉蛋白在酸性条件下可自发插入脂单层.科学通报,1999,44(16):1740-1743
[22]戴荣禧,徐国江,刘莲英,等.天花粉蛋白对滋养层细胞转移损伤机制的研究.实验生物学报,1993,26:411-416
[23]许良中.实用肿瘤病理方法学(第一版).上海:上海医科大学出版社,1997.578-595
[24]Darzynkiewicz Z,Juan G,Li X,et al.Cytometryin cell necrobiology:analysis of apoptosis and accidental cell death(Necrosis).Cytometry,1997,27(1):1-20
[25]ZHANG SHU,HU MEIJIE,WU YUXIN,et al.Chinese Journal of Digestive Diseases,2001,2:125-128
[26]张曙,胡梅洁.天花粉蛋白诱导的胃癌细胞凋亡与bcl-2表达下降有关.中华消化杂志,2000,20(6):380
[27]Hu MJ,Zhang S,Wu YX,Jiang SH.Synergic cytotoxicity to gastric cancer cells by combined use of trichosanthin and recombinant interferon alpha-2b.J Shanghai Second Med University,2000,12:44-47
[28]Luo WD,Ren.CM,Zhu M,et al.Study on NB4 cell apoptosis induced by trichosanthin.Zhongguo Shi Yan Xue Ye Xue Za Zhi,2005,13:278-281
[29]孔梅,柯一保,周关云,等.天花粉蛋自诱发白血病细胞K562凋亡的研究.实验生物学报,1998,31:233-243
[30]黄益玲,黄利鸣,石新兰,等.天花粉蛋白对人宫颈癌HeLa细胞增殖和细胞凋亡的影响.中国药理学通报,2005,21:253-254
[31]毕黎琦,李洪军,张玉华,等.中药天花粉蛋白对黑色素瘤细胞凋亡及细胞周期的影响.中国中西医结合杂志,1998;18:35-37
[32]李振红,陆阳,刘晶星.天花粉化学成分与药理活性.国外医药(植物药分册),2003,18(1):1-4
[33]Li MX,Yeung HW,Pan LP,et al.Trichosanthin,a potent HIV-Ⅰinhibitor can cleave supercoiled DNA in vitro.Nucleic Acids Res,1991,19(22):6309-6312
[34]何肾辉,曾耀英,孙荭,等.天花粉蛋白诱导人类白血病细胞株HL-60细胞凋亡的研究.中国病理生理杂志,2001,17(3):200
[35]魏明卉,李英,刘征.核糖体失活蛋白及研究进展.南都学坛(自然科学版),2001,21(6):64-67
[36]胡红雨.鲁子贤.1H NMR法研究天花耢蛋白催化5'-AMP糖苷酶的裂解.科学通报,1994,39:1720-1722
[37]Meguire WL.Hormone receptors:their role in predicting prognosis and response to endocrine therapy.Semin Oncol,1978,5:428-433
[38]Sommer S,Fuqua SA.Estrogen receptor and breast cancer.Semin Cancer Biol,2001,11:339-352
[39]Lapidus RG,Nass SJ,Davidson NE.The loss of estrogen and progesterone receptor gene expression in human breast cancer.J Mamm Gland Biol Neoplasia,1998,3:85-94
[40]Ottaviano YL,Issa JP,Pad FF,et al.Methylation of the estrogen receptor gene CpG island marks loss of estrogen receptor expression in human breast cancer cells.Cancer Res,1994,54(10):2552-2555
[41]Ferguson AT,Lapidus RG,Baylin SB,et al.Demethylation of the estrogen receptor gene in estrogen receptor-negative breast cancer cells can reactivate estrogen receptor gene expression.Cancer Research,1995,55(11):2279-2283
[42]韩明强,黄韬,田元.雌激素水平对三苯氧胺抑制乳腺癌细胞的体外研究.中国普外基础与临床杂志,2005,12(6):589-590,594
[43]Matins-Guiu X,Catasus L,Bussaglia E,et al.Molecular pathology of endometrial hyperplasia and carcinoma.Hum Pathol,2001,32(6):569-577
[44]Becker K,Kellerq,Hoefler H.The use molecular biology in diagnosis and prognosis of gastric cancer.Surg Oncol,2000,9(1):5-11
[45]Nagane Y,Utsugisawa K,Tobgi H.PCR amplification in bisulfite methylcytosine mapping in the GC-rich promoter region of a myloid precursor protein gene in autopsy human brain.Brain Res Prow,2000,5(2):167-171
[46]Chide Y,Ishizaki M,Nakazawa M.Expression and methylation of the beta-subunit gene of prolyl 4-hydroxylase:in erythrocytes,tendon and cornea of chickembryos.Connect Tissue Res,1992,28(3):191-204
[47]Asch BB,Barcellos-Hoff MH.Epigenetics and breast cancer.J Mammary Gland Biol Neoplasia,2001,6:151-152
[48]Cameron EE,Bachman KE,Myohanen S,et al.Synergy of demethylation and histone deacetylase inhibition in the re-expression of genes silenced in cancer.Nat Genet,1999,21:103-107
[49]Donaghue C,Westley BR,May FE.Selective promoter usage of the human estrogen receptor-alpha gene and its regulation by estrogen.Mol Endocrinol,1999,13:1934-1950
[50]Osborne C K.Steroid hormone receptors in breast cancer management.Breast Cancer Research and Treatment,1998,51(3):227-238
[51]Ali S,Coombes RC.Estrogen receptor alpha in human breast cancer:occurrence and significance.J Mammary Gland Biol Neoplasia,2000,5(3):271-281
[52]Hayashi SI,Eguchi H,Tanimoto K,et al.The expression and function of estrogen receptor alpha and beta in human breast cancer and its clinical application.Endocr Relat Cancer,2003,10(2):193-202
[53]Roodi N Bailey LR,Kao WY,et al.Estrogen receptor gene analysis in estrogen receptor-positive and receptor-negative primary breast cancer.J Natl Cancer Inst,1995,87(6):446-451
[54] Watts CK, Handel ML, King RJ, et al. Oestrogen receptor gene structure and function in breast cancer. J Steroid Bio chem Mol Biol, 1992,41(3-8): 529-536
[55] Iwase H, Kobayashi S, Iwata H, et al. Molecular analysis of the estrogen receptor(ER) gene in association with ER negativity in breast cancer. Gan To Kagaku Ryoho. 1996,23(Suppl 1): 61-65
[56] Dowsett M, Dafada A, Chan CM, et al. Oestrogen receptor mutants and variants in breast cancer. Eur J Cancer, 1997, 33(8): 1177- 1183
[57] Stoner M, Saville B,Wormke M, et al. Hypoxia induces proteasome dependent degradation of estrogen receptor alpha in ZR275 breast cancer cells. Mol Endocrinol, 2002,16:2231-2242
[58] Roger P, Daures JP, Maudelonde T, et al. Dissociated over expression of cathepsin D and estrogen receptor alpha in preinvasive mammary tumors. Hum Pathol, 2000,31:593-600
[59] Hewitt SC, Bocchinfuso WP, Zhai J, et al. Lack of ductal development in the absence of functional estrogen receptor alpha delays mammary tumor formation induced by transgenic expression of ErbB2/neu. Cancer Res, 2002, 62(10): 2798-2805
[60] Lapidus RG» Nass SJ, Butash KA. et al. Mapping of ER gene CpG island methylation-specific polymerase chain reaction. Cancer Res. 1998, 58(12): 2515- 2519
[61] Xiong Z, Laird PW. COBRA: a sensitive and quantitative DNA methylation assay-Nucleic Acids Res, 1997,25(12): 2532-2534
[62] Suzuki H, Itoh F, Toyota M, et al. Quantitative DNA methylation analysis by fluorescent polymerase chain reaction single-strand conformation polymorphism using an automated DNAs equence. Electrophoresis, 2000,21(5): 904-908
[63] Gonzalgo ML, Jones PA. Rapid quantitation of methylation differences at specific sites using methylation-sensitive single nucleotide primer extention(Ms-SnuPE). Nucleic Acids Res, 1997,25(12): 2529-2531
[64] Bianco T, Hussey D, Dobrovic A. Methylation-sensitive single strand conformation analysis(MS-SSCA): A rapid method to screen for and analyze methylation. Hum Mutat, 1999,14(4): 289-293
[65] Eads CA, Danenberg KD, Kawakami K, et al. MethyLight: a high-throughput assay to measure DNA methylation. Nucleic Acids Res, 2000,28(8): E32
[66] Esteller M, Catasus L, Matias-Guiu X, et al.hMLHl promoter hypermethylation is an early event in human endometrial tumorigenesis. Am J Pathol, 1999,155:1767-1772
[67] Jones PA, Laird PW. Cancer epigenetics comes of age. Nat Genet, 1999, 21(2): 163-167
[68] Herman JG, Graff JR, Myohanen S, et al. Methylation-specific PCR: a novel PCR assay for methylation status of CpG islands. Proc Natl Acad Sci USA, 1996, 93(18): 9821-9826
[69] Singal R, Ginder GD. DNA methylation. Blood, 1999, 93:4059-4070
[70] Hortobagyi GN. Treatment of breast cancer. N Engl J Med, 1998, 339: 974- 984
[71] Weigel RJ, deConinck EC. Transcriptional control of estrogen receptor in estrogen receptor-negative breast carcinoma. Cancer Res 1993,53: 3472-3474
[72] Wolffe AP,Matzke MA. Epigenetics: regulation through repression. Science, 1999,286(5439): 481-486
[73] Zhao C, Lam EW , SuntersA. et al. Expression of estrogen recptor beta isoformsin normal breast epithelial cells and breast cancer : regulation by methylation. Oncogene, 2003, 22(48): 760007606
[74] Jones PA, Laird PW. Cancer epigenetics comes of age. Nat Genet, 1999, 21: 163-167
[75] Baylin SB, Herman JG. DNA hypermethylation in tumorigenesis: epigenetics joins genetics. Trend Genet, 2000,16: 1680174
[76] Momparler RL, Bovenzi V. DNA methylation and cancer. J Cell Physiol, 2000, 183(2): 45-54
[77] Bird AP CPC-rich islands and the function of DNA methylation. Nature, 1986, 321 (6067) : 206-213
[78] Dominguez G, Silva J, Garcia JM, et al. Prevalence of aberrant methylation of pl4ARF over p16INK4a in some human primary tumors. Mutat Res Fundam Mol Mech Mutagen, 2003,530 (1) : 9-17
[79] Nicolai M, Marco B, Frank R, et al. Hypermethylation and histone deacetylation lead to silencing of the maspin gene in human breast cancer. Biochemical and Biophysical Research Communications, 2002, 297(1): 125-128
[80]Christman JK.5-Azacytidine and 5-aza-2'-deoxycytidine as inhibitor of DNA methylation:mechanistic studies and their implications for cancer therapy.Oncogene,2002,21:5483
[81]温国林,陈晓光.DNA甲基化及其抑制剂研究进展.国外医学遗传学分册,2001,24(4):180-184
[82]Rochefort H,Glondu M,Sahla ME,et al.How to target estrogen receptor-negative breast cancer? Endocr Relat Cancer,2003,10(2):261-266
[83]Varga Z,Zhao J,Ohlschlegel C,et al.Preferential HER-2/neu overexpression and/or amplification in aggressive histological subtypes of invasive breast cancer.Histopathology,2004,44(4):332-338
[84]Laurent MV,Lucas A,Huttler S,et al.Procathepsin D interacts with prosapostn in can c cells but its internalization is not mediated by LDL receptor- related protein.Exp Cell Res,2002,277(2):210-219
[85]Shibata A,Nagaya T,Imai T,et al.Inhibition of NF-kappaB activity decreases the VEGF mRNA expression in MDA-MB-231 breast cancer cells.Breast Cancer Res Treat,2002,73(3):237-43
[86]Hirvonen R,Talvensaari MA,Paakko P,et al.Matrix metalloproteinase-2(MMP-2) in T_(1-2)N_0 breast carcinoma.Berast Cancer Res Treat,2003,77(1):85-91
[87]Franke H R,Kole S,Ciftci Z,et al.In vitro effects of estradiol,dydrogesterone,tamoxifen and cyclophosphamide on proliferation vs.death in human breast cancer cells.Cancer Letters,2003,190(1):113-118
[88]Shao Z,Jiang M,Yu L,et al.Estrogen receptor-negative breast cancer cells transfected with estrogen receptor exhibit decreased tumour progression and sensitivity to growth inhibition by estrogen.Chin Med Sci J,1997,12(1):11
[89]Tsao SW,Ng TB,Yeung HW.Toxicities of trichosanthin and alpha-momorcharin,abortifacient proteins from Chinese medicinal plants,on cultured tumor cell lines.Toxicon,1990,28:1183-1192
[90]汪钊猷主编.天花粉蛋白.上海:科学出版社,1990.4-7
[91]孙健,吴志全,李雁,等.天花粉蛋白与重组人干扰素α-1b协同抑制人肝细胞癌的实验研究.中国临床医学,2003,10(3):278-280
[92]徐振武,吴裕忻,陈陵际,等.天花粉蛋白对荷人结肠癌SW-1116裸鼠的体内实验研究肿瘤学杂志,1997,3(2):110-111
[93]Ln JB,LiMY,et al.Detenmination of the binding epitope for anti-trichosanthin monochonal antibodyT(8)C(12).Sheng Wu Hua Xue Yu Sheng Wu Wu Li Xue Bao,1995,30:137-141
[94]唐文皓,吴志全,等.樊嘉天花粉蛋白对荷人肝癌裸小鼠的抑癌作用.中华肝胆外科杂志,2003,9(4):228-230
[95]Ke YB,Chen J K,Hie HL,et al.Structure-function relationship of trichosanthin.Life Sci,1997,60:465-472
[96]张劲松,刘望夷.天花粉蛋白在真核细胞核糖体28S rRNA上的作用位点.生物化学与生物物理进展,1992,19:131
[97]田维毅,马春玲,白惠卿.天花粉及其组份对小鼠NK细胞杀伤活性的影响.贵州医学杂志,2001,25:982-984
[98]田维毅,马春玲,白惠卿.天花粉及其组份对小鼠NK细胞杀伤活性的影响.贵州中医,2001,25(11):289
[99]周广宇,汪丽,毕黎琦,等.天花粉蛋白对人PBMC分泌细胞因子水平及离子通道的影响.中国免疫学杂志,2001,17:650-652
[100]郭峰,刘小平,徐瑛.天花粉对小鼠艾氏腹水癌的疗效和对红细胞免疫功能的影响.中西医结合杂志,1989,9:418-420
[101]骆和生,王建华,邓文龙,等.中药方剂的药理与临床研究进展.广州:华南理工大学出版社,1991.78
[102]Myer RA,Sergios PA,Coonan K,et al.Trichosanthin treatment of HIV-induced immunedys regulation.Eur J Invest 1992,22(2):113
[103]奚正德,马宝骊,曹鹤年,等.天花粉蛋白羟胺裂解片段制备及其对补体的激活作用.中国免疫学杂志,1997,13(1):47
[104]郭峰,刘小平,徐瑛.天花粉对小鼠艾氏腹水癌的疗效和对红细胞免疫功能的影响.中西医结合杂志,1989,9(7):418-420
[105]徐瑛,郭峰,叶天星。天花粉治疗小鼠艾氏腹水癌对红细胞增强吞噬作用的研究.肿瘤,1990,10(5):224
[106]邬伟秀,郭峰.天花粉对血液肿瘤患者红细胞免疫功能影响的实验研究.上海免疫学杂志,1995,5(3):156
[107]洪建,富赛里,陆佩华,等.天花粉蛋白对T细胞活化的抑制作用与信号传导.中华微生物学和免疫学杂志,1998,18(2):111
[108]胡梅洁,张曙.天花粉蛋白与重组干扰素α-2b联用对荷人胃腺癌SGC-7901 裸小鼠的抑癌作用.Shanghai Med J,1998,21(8):456-457
[109]Jones PA,Taylor SM,Mohandas T,et al.Cell cycle-specific reactivation of an inactive X-chromosome locus by 5-aza-deoxycytidine.Proc Natl Acad Sci USA,1982,79:1215-1219
[110]Yoshiura K,Kanai Y,Ochiari A,et al.Silencing of the E-cadherin invasion-suppressor gene by CpG methylation in human carcinomas.Proc Natl Acad Sci USA,1995,92:7416-7419
[111]Au WY,Fung A,Man C,et al.Aberrant p15 gene promoter methylation in therapy-related myelodyspiastic syndrome and acute myeloid leukaemia:clinicopathological and karyotypic associations.Br J Haematoi,2003,120(6):1062-1065
[112]Lemaire M,Momparler LF,Farinha NJ,et al.Enhancement of antineoplastic action of 5-aza-2'-deoxycytidine by phenylbutyrate on L1210 leukemic cells.Leuk Lymphoma,2004,45(I):147-154
[113]Miyamoto K,Ushijima T.DNA methylation and cancer-DNA methylation as a target of cancer chemotherapy.Gan To Kagaku Ryoho,2003,30(13):2021-2029
[114]Silverman LR,Holland JF,Davis RB,et al.Azacytidine induces trilineage responses in patients with myelodysplastic syndromes:the Cancer and Leukemia Group B experience.Hematologica,1991,76:42
[115]Jeffrey G,Zella RZ,Richard KS,et al.Treatment of myelodysplastic syndromes with 5-azacytidine.Leukemia Research,2002,26:893-897
[116]Tamura T,Kataoka A,Shu LY.et al.An in vitro screening method for DNA cytosine-C5-methylase inhibitor.Nat Prod Lett,2002,16(1):25
[117]Jeong LS,Yoo S J,Lee KM,et al.Design,synthesis,and biological evaluation of fluoroneplanocin A as the novel mechanism-based inhibitor of S-adenosylhomocysteine hydrolase.J med Ctiem,2003,46(2):201-203
[118]黄利鸣,邢学森,黄益玲.p16基因甲基化与Hela细胞凋亡相关性的研究.中国肿瘤临床,2005,32(13):721-724
[119]RX Dai,GL Xu,LY Liu,et al.Study on injury-mechanism of trichosanthin on trophoblast cells and choriocarcinoma cells in culture.Shiyan Sshengwu Xuebao,1993,26(4):411-427
[120]胡梅洁,吴云林,张曙,等.消化病与内镜(第一版).上海:上海科技文 献出版社.1998.281-284
[121]马宝骊,陈汐,杨黎明.天花粉蛋白类过敏反应分子机理的研究.医学研究通讯,1993,22(5):25
[122]黄健,金毓翠.天花粉蛋白宫颈注射引产使用地塞米松减少副反应的分析.现代妇产科进展,1994,3:227-228
[123]Byers VS,Levin AS,Malvino A,et al.A phase Ⅱstudy ofeffect of addition of trichosanthinto zidovudine in patients with HIV disease and failing antiretroviral agents.AIDS Res Hum Retroviruses,1994,10(4):413-420
[124]张淑景,金毓翠,于金德,等.天花粉引产对孕妇心、肝、肾功能影响的进一步研究.生殖与避孕,1994.14:19-24
[125]张素胤,张家骝.8种抗癌药物对裸小鼠肾包膜下接种的人肺腺癌移植瘤(LAX-83)生长的影响.中国药理学报,1989,15(5):450-453
[126]王茂钦,李莉,王洁,等.天花粉蛋白致豚鼠全身过敏反应的实验研究.安徽中医学院学报,1999,18:47-49
[127]徐振武,吴裕忻,陈陵际,等.天花粉蛋白抗结肠癌体内实验研究.海峡药学,1997,9:12-13
[128]时伟红,宋今丹.含天花粉蛋白的抗人大肠癌免疫毒素制备及其细胞毒性研究.中国医科大学学报,1996,25(2):111-113
[1]Wolffe A P,Matzke M A.Epigenetics:regulation through repression.Science,1999,286(5439):.481-486
[2]王树人.基因的表达与Epigenetics.四川生理科学杂志.2006,28(1):29-33
[3] J aenisch R ,Bird A. Epigenetic regulation of gene expression : how the genome integrates intrinsic and environmental signals. Not Rev Genet ,2003 ,33 :245-254
[4] Matias K P. Study of epigenetic changes leads to treatment advances in adult leukemias. Oncolog ,2003 ,48(1): 1-4
[5] Okano M,Bell ,DW,Haber DA ,Li E. DNA methyltransferases Dnmt3a and Dnmt3b are essential for de novo methylation and mammalian development. Cell ,1999,99(3):247-257
[6] Reik W, Dean W, Walter J. Epigenetic reprogramming in mammalian development. Science, 2001,293(5532): 1089-1093
[7] Rush LJ, Plass C. Restriction landmark genomic scanning for DNA methylation in cancer :past ,present ,and future applications. Anal Biochem,2002 ,307(2): 191 -201
[8] Robertson KD Jones PA. DNA methylation : past ,present and future directions. Carcinogenesis ,2000,21 (3) :461-467
[9] Peter A J , Daiya T. The role of DNA methylation in mammalian epigenetics. Science, 2001,293(5532): 1068-1070
[10] Ichimura T , Watanabe S , Sakamoto Y, et al. Transcriptional repression and heterochromatin formation by MBD1 and MCAF/AM family proteins. J Biol Chem, 2005,280(14): 13928-13935
[11] Tamura G, Yin J , Wang S , et al. E-Cadherin gene promoter hypermethylation in primary human gastric carcinomas. J Natl Cancer Inst, 2000,92(7):569-573
[12] Manini DN ,Rodenhiser DI ,Ainsworth PJ , et al. CpG methylation within the 5'regulatory region of BRCAL ,genies tumor specific and includes a putative CREB binding site. Oncogene, 1998,16(9): 1161 - 1169
[13] Clark SJ ,Harrison J ,Molloy PL. Spl binding is inhibited by CpG methylation. Gene,1997,195 (1) :67-71
[14] Singal R, Ginder GD. DNA methylation. Blood, 1999,93(12) :4059-4070
[15] Nan X,Ng HH ,Johnson CA , et al . Transcriptional repression by the met hyl CpG binding protein MeCP2 involves a histonedeacetylase complex. Nature ,1998 ,393 (6683):386-389
[16] Ng HH , Zhang Y, Hendrich B, et al . MBD2 is a transcriptional repressor belonging to the MeCPl histone deacetylase complex. Nat Genet, 1999,23(1): 58-61
[17] Momparler RL ,Bovenzi V. DNA methylation and cancer. J Cell Physiol, 2000, 183(2): 145-154
[18] Jones PA, Laird PW. Cancer epigenetics comes of age. Nat Genet,1999,21(2): 163-167
[19] Hsieh CL. Evidence t hat protein binding specifies sites of DNA demethylation. Mol Cell Biol,1999,19(1) :46-56
[20] Wolffe AP ,Jones PL ,Wade PA. DNA demethylation. Proc Natl Acad Sci USA, 1999,96(11): 5894-5896
[21] Jost J P ,Siegmann M ,Sun L ,et al . Mechanisms of DNA demethylation in chicken embryos. Purification and properties of a 5-methylcytosine-DNA glycosylase.J Biol Chem,1995,270(17): 9734-9739
[22] Zhu B ,Zheng Y ,Hess D ,et al. 5-methylcytosine-DNA glycosylase activity is present in a cloned G/T mismatch DNA glycosylase associated with the chicken embryo DNA demethylation complex. Proc Natl Acad Sci USA,2000,97(10) :5135-5139
[23]朱卫国. DNA甲基化, 基因调控和癌症. 世界华人消化杂志, 2002, 10(6): 680-683
[24] Grady WM, Willis S , Guilford PJ, et al. Methylation of the CDH1 promoter as the second genetic hit in hereditary diffuse gastric cancer.Nat Genet, 2000,26(1): 16-17
[25] Jones PA , Takai D. The role of DNA methylation in mammalian epigenetics. Science, 2001,293(5532):1068- 1070
[26] Jones PA ,Baylin SB. The fundamental role of epigenetic events in cancer. Nat Rev Genet,2002,3(6):415-428
[27] Lee WH ,Morton RA ,Epstein J I ,Brooks JD ,Campbell PA ,Bova GS , et al . Cytidine methylation of regulatory sequences near the piclass glutathione S-transferase gene accompanies human prostatic carcinogenesis. Proc Natl Acad Sci USA, 1994,91(24): 11733-11737
[28] Leone G,Teofili L ,Voso MT, et al. DNA methylation and demethylating drugs in myelodysplastic syndromes and secondary leukemias. Haematologica, 2002, 87(12): 1324-1341
[29] Robertson KD. DNA methylation and human disease. Nat Rev Genet, 2005, 6(8):597-610
[30] Li QL , Ito K, Sakakura C , et al . Causal relationship between the loss of RUNX3 expression and gastric cancer. Cell, 2002 ,109(1): 113-124
[31]李燕妮,张维铭.促甲基化反应的研究进展.国际检验医学杂志,2006,27(6):527-528,531
[32]Butcher DT,Mancini-DiNardo DN,Archer TK,et al.DNA binding sites for putative methylation boundaries in the unmethylated region of the BRCA1 promoter.Int J Cancer,2004,111(5):669-678
[33]Etoh T,Kanai Y,Ushijima S,et al.Increased DNA methyltransferase 1(DNMTI) protein expression correlates significantly with poorer tumor differentiation and frequent DNA hypermethylation of multiple CpG islands in gastric cancers.Am J Pathol,2004,164(2):689-699
[34]Nagai M,Nakamura A,Makino R,et al.Expression of DNA(5-cytosin)-methyltransferases (DNMTs) in hepatocellular carcinomas.Hepatol Res,2003,26(3):186-191
[35]Paz M F,Fraga M F,Avila S,et al.A systematic profile of DNA methylation in human cancer cell lines.Cancer Res,2003,63(5):1114-1121
[36]Yang B,Guo M,Herman J G,et al.Aberrant promoter methylation profiles of tumor suppressor genes in heaptocellular carcinoma.Am J Pathol,2003,163(3):1101-1107
[37]Esteller M,Herman JG.Cancer as an epigenetic disease:DNA methylation and chromatin alterations in human tumours.J Pathol,2002,196(1):127
[38]Wong N,Lam W C,Lai P B,et al.Hypomethylation of chromosome 1heterochromatin DNA correlates with qarm copy gain in human heaptocellular carcinoma.Am J Pathol,2001,159(2):465-471
[39]Nakagawa T,Kanai Y,Ushijima S,et al.DNA hypomethylation on pericentromeric satellite regions significantly correlates with loss of heterozygosisty on chromosome 9 in urothelial heteozygosity on chromosome 9 in urothelial carcinomas.J Urol,2005,173(1):243-246
[40]Rusmintratip V,Riggs AD,Sowers LC.Examination of the DNA substrate selectivity of DNA cytosine methyltransferases using mass tagging.Nucleic Acids Res,2000,28(18):3594-3599
[41]Florl AR,Lower R,Schmitz Drager BJ,et al.DNA methylation and expression of LINE21 and HERV2Kprovirus sequences in urothelial and renal cell carcinomas.Br J Cancer,1999,80(9):1312-1321
[42]Feinberg AP.Imprinting of a genomic domain of 11p15 and loss of imprinting in cancer:an introduction.Cancer Res,1999,59(7Suppl):1743-1746
[43]Kokalj-Vokac N,Almeida A,Viegas-Pequignot E,et al.Specific induction of uncoiling and recombination by azacytidine in classical satellite-containing constitutive heterochromatin.Cytogenet Cell Genet,1993,63(1):11-15
[44]吕斌斌,彭剑雄.DNA甲基化与基因沉默及肿瘤.国际检验医学杂志,2006,27(1):52-54,57
[45]Tao L,Yang S,Xie M,et al.Hypomethylation and overexpression of c-jun and c-myc protooncogenes and increased DNA methyltransferase activity in dichloroacetic and trichloroacetic acid-promoted mouse liver tumors.Cancer Lett,2000,158(2):185-193
[46]Jang S J,Soria JC,Wang L,et al.Activation of melanoma antigen tumor antigens occurs early in lung carcinogenesis.Cancer Res,2001,61(21):7959-7963
[47]Feng Z,Hu W,Rom WN,et al.N-hydroxy-4-aminobiphenyl-DNA binding in human p53 gene:sequence preference and the effect of C5 cytosine methylation.Biochemistry,2002,41(20):6414-6421
[48]Florl AR,Steinhoff C,Muller M,et al.Coordinate hyper- methylation at specific genes in prostate carcinoma precedes L1NE-1 hypomethylation.Br J Cancer,2004,91(5):985-994
[49]Seiandrello G,Caradonna F,Mauro M,et al.Arsenic-induced DNA hypomethylation affects chromosomal instability in mammalian cells.Carcinogenesis,2004,25(3):413-417
[50]Ehrlich M.The ICF syndrome,a DNA methyltransferase 3B deficiency and immunodeficiency disease.Clin Immunol,2003,109(1):17-28
[51]Gaudet F,Hodgson JG,Eden A,et al.Induction of Tumors in Mice by Genomic Hypomethylation.Science,2003,300(5618):489-492
[52]Wu J,lssa J,Hermen J,et al.Expression of an exogenous eukaryotic DNA methyltransferase gene induces transformation of NIN3T3 ceils.Proc Natl Acad Sci USA,1993,90(19):8891-8895
[53]Oakeley EJ,Schmitt F,Jost JP.Quantification of 5-methylcytosine in DNA by the chloroacetaldehyde reaction.Biotechniques,1999,27(4):744-746
[54]Wainfan E,Dizik M,Stender M,et al.Methyl group in carcinogenesis:effects on DNA methylation and gene expression. Cancer Res, 1992,52(7Suppl): 2071s-2077s
[55] Richard L , Robey R , Sandor V , et al. Inhibitors of histone deacetylation , Depsipeptide (FR901228) , in the treatment of peripheral and cutaneous T-cell lymphoma: a case in report. Blood,2001,98(9):2865-2868
[56] Urnov FD , Wolffe AP. Chromatin remodeling and transcriptional activation: the cast (in order of appearance). Oncogene, 2001,20(24 Rev 3):2991-3006
[57] Li Q , Ahuja N , Burger PC , et all Methylation and silencing of the Thrombospondin-1 promoter in human cancer. Oncogene, 1999,18(21): 3284 -3289
[58] Pogribny I, Yi P , James SJ , et al. A sensitive new method for rapid detection of abnormal methylation patterns in global DNA and within CpG islands. Biochem Biophys Res Commun, 1999,262(3): 624-628
[59] Rein T , Natale DA , Gartner U , et al. Absence of an unusual"densely methlated island"at the hamster ori-beta. JBiol Chem, 1997,272(15): 10021 - 10029
[60] Yamaguchi Y., Wada T., Suzuki F., et al. Casein kinase II interacts with the bZIP domains of several transcription factors. Nucleic Acids Res, 1998, 26(16): 3862- 3864
[61] Herman JG, Graft JR , Myohanen S , et al. Methylation-specific PCR: a novel PCR assay for methylation status of CpG islands. Proc Natl Acad Sci USA, 1996 , 93(18):9821-9826
[62] Redner RL , Wang J , Liu JM. Chromatin remodeling and leukemia: new therapeutic paradigms. Blood, 1999,94(2): 417-428
[63] Rand K, Qu W, Ho T, et al. Conversion-specific detection of DNA methylation using real-time polymerase chain reaction (ConLight-MSP) to avoid false positives. Methods, 2002 ,27(2): 114-120
[64] Yan PS , Chen CM, Shi H , et al. Dissecting comples epigenetic alterations in breast cancer using CpG island microarrays. Cancer Res, 2001,61(23):8375-8380
[65] Gonzalgo ML , Jones PA1 Rapid quantitation of methylation differences at specific sites using methylation-sensitive single nucleotide primer extension (Ms- NuPE). Nucleic Acids Res, 1997,25(12): 2529-2531
[66] El-Maarri D , Herbiniaux U , Walter J , et al. A rapid quantitative non- adioactive bisuifite-SnuPE-IP RP HPLC assay for methylation analysis at specific CpG sites . Nucleic Acids Res, 2002 , 30(6):e25
[67]Grunstein M.Histone acetylation in chromatin structure and transcription.Nature,1997,389(6649):349-352
[68]Oefner Pi,Bonn GK,Huber CG,et al.Comparative study of capillary zone electrophoresis and high-performance liquid chromatoguaphy in the analysis of oligonucleotides and DNA.J Chromatogrl 1992,625(2):331-401
[69]邓大君,邓国仁,吕有勇,等.变性高效液相色谱法检测CpG岛胞嘧啶甲基化.中华医学杂志,2001,80(2):158-161
[70]Banmer A,Wiedemann U,HergersbergM,et al.A novel MSP-DHPLC method for the investigation of the methylation status of imprintedgenes enables the molecular detection of low cell mosaicisms.Hum Mutat,2001,17(5):423-430
[71]Slater H,Brtmo D,Ren H,et al.Improved testing for CMT1A and HNPP using multiplex ligation-dependent probe amplification(MLPA) with rapid DNA preparations:comparison withthe interphase FISH method.Hum Mutat,2004,24(2):164-171
[72]Slater HR,Bruno DL,Ren H,et al.Rapid,high throughput prenatal detection of aneuploidy using a novel quantitative method(ML PA).J Med Genet,2003,40(12):907-912
[73]Worsham MJ,Pals G,Schouten J P,et al.Delineating genetic pathways of disease progression in head and neck squamous cell carcinoma.Arch Otolaryngol Head Neck Surg,2003,129(7):702-708
[74]Schouten J P,Mcelgunn CJ,Waaijer R,et al.Relative quantification of 40nucleic acid sequences by multiplex ligation-dependent probe amplification.Nucleic Acids Res,2002,30(12):e57
[75]Nygren AO,Ameziane N,Duarte HM,et al.Methylation-Specific MLPA(MS-MLPA):simultaneous detection of CpG methylation and copy number changes of up to 40 sequences.Nucleic Acids Res,2005,33(14):e128
[1]Palmisano W A,Divine K K,Saccomanno G,et al.Predicting lung cancer by detecting aberrant promoter methylation in sputum[J].Cancer Res,2000,60(21):5954-5958
[2]Cavalli L R,Urban C A,Dai D,et al.Genetic and epigenetic alterations in sentinel lymph nodes metastatic lesions compared to their corresponding primary breast tumors[J].Cancer Genet Cytogenet,2003,146(1):33-40
[3]Primeau M,Gagnon J,Momparler R L.Synergistic antineoplastic action of DNA methylation inhibitor 5-AZA-2'-deoxycytidine and histone deacetylase inhibitor depsipeptide on human breast carcinoma cells[J].Int J Cancer,2003,103(2):177-184
[4]Yang X,Yan L,Davidson N E.DNA methylation in breast cancer[J].Endocr Relat Cancer,2001,8(2):115-127
[5]Herman J G,Baylin S B.Promoter region hypermethylation and gene silencing in human cancer[J].Curr Top Mierobiol Immunol,2000,24(9):35-54
[6]Woodcock D M,Linseumeyer M E,Doherty J P,et al.DNA methylation in the promoter region of the p16(CDKN2PMTS21PINK4A) gene in human breast tumours[J].Br J Cancer,1999,79(2):251-256
[7]王志伟,易静,陈玉英,等.乳腺癌中p16基因甲基化及其与临床病理学特征的关系[J].中华实验外科杂志,2002,19(1):81
[8] Takai D, Jones P A. Comprehensive analysis of CpG islands in human chromosomes 21 and 22[J]. Proc Natl Acad Sci USA, 2002,99(6): 3740-3745
[9] Jones P A, Laird P W. Cancer epigenetics comes of age[J]. Nat Genet, 1999 , 21(2): 163-167
[10] Yang X, Ferguson A T, Nass S J, et al. Transcriptional activation of estrogen receptor a in human breast cancer cells by histone deacetylase inhibition[J]. Cancer Res, 2000,60(24): 6890-6894
[11] Ferguson A T, Evron E, Umbricht C B, et al. High frequency of hypermethylation at the 14-3-3 o locus leads to gene silencing in breast cancer[J]. Proc Natl Acad Sci USA, 2000,97(11): 6049-6045
[12] Celis A, Rasmussen H H, Celis P, et al. Short-term culturing of low-grade superficial bladder transitional cell carcinomas leads to changes in the expression levels of several proteins involved in key cellular activities[J].Electrophoresis, 1999, 20(2): 355-361
[13] Ottaviano Y L, Issa J P, Parl F F, et al. Methylation of the estrogen receptor gene CpG island marks loss of estrogen receptor expression in human breast cancer cells[J]. Cancer Res, 1994, 54(10): 2552-2555
[14] Ferguson A T, Vertino P M, Spitzner J R, et al. Role of estrogen receptor gene demethylation and DNA methyltransferase- DNA adduct formation in 5-aza-2'-deoxycytidine-induced cytotoxicity in human breast cancer cells[J]. J Biol Chem, 1997,272(51): 32260-32266
[15] Onate S O, Tsai S Y, Tsai M J, et al. Sequence and characterization of a coactivator for the steroid hormone receptor superfamily[J]. Science, 1995, 270(5240): 1354-1356
[16] Du Y, Carling T, Fang W, et al. Hypermethylation in human cancers of the RIZ1 tumor suppressor gene, a member of a histone/protein methyltransferase superfamily [J]. Cancer Res, 2001, 61(22): 8094-8099
[17] Graff J R, Gabrielson E, Fujii H, et al. Methylation patterns of the E-cadherin 5' CpG island are unstable and reflect the dynamic, hetero- geneous loss of E-cadherin expression during metastatic progression[J]. J Biol Chem, 2000 ,275(4): 2727-2732
[18] Chen C L, Liu S S, Ip S M, et al. E-cadherin expression is silenced by DNA methylation in cervical cancer cell lines and tumours [J]. Eur J Cancer , 2003, 39(4): 517-523
[19]Nass S J,Herman J G,Gabrielson E,et al.Aberrant methylation of the estrogen receptor and E-cadherin 5'CpG islands increases with malignant progression in human breast cancer[J].Cancer Res,2000,60(16):4346-4348
[20]Toyooka K O,Toyooka S,Virmani A K,et al.Loss of expression and aberrant methylation of the CDH13(H-eadhedn) gene in breast and lung carcinomas[J].Cancer Res,2001,61(11):4556-4560
[21]Chen L M,Chai K X.Prostasin serine protease inhibits breast cancer invasiveness and is transcriptionally regulated by promoter dna methy- lation[J].Int J Cancer,2002,97(3):323-329
[22]Catteau A,Harris W H,Xu C F,et al.Methylation of the BRCA1 promoter region in sporadic breast and ovarian cancer;correlation with disease characteristics[J].Oncogene,1999,18(11):1957-1965
[23]Esteller M,Corn P G,Urena J M,et al.Inactivation of glutathione Stransferase P1 gene by promoter hypermethylation in human neoplasia[J].Cancer Res,1998,58(20):4515-4518
[24]李云青,张恩仲,肖翠英,等.乳腺癌患者BRCA1基因启动子区甲基化模式的初步研究[J].癌症,2001,20(10):1057-1060
[25]Magdinier F,Billard L M,Wittmann G,et al.Regional methylation of the 5'end CpG island of BRCA1 is associated with reduced gene expression in human somatic cells[J].FASEB J,2000,14(11):1585-1594
[26]Fujii H,Biel M A,Zhou W,et al.Methylation of the HIC-1 candidate tumor suppressor gene in human breast cancer[J].Oncogene,1998,16(16):2159-2164
[27]Esteller M,Sanchez-Cespedes M,Rosell R,et al.Detection of aberrant promoter hypermethylation of tumor suppressor genes in serum DNA from non-small cell lung cancer patients[J].Cancer Res,1999,59(1):67-70
[28]黄世杰.癌的表观遗传性治疗:过去,现在和将来[J].国外医学药学分册:2006,33(5):342-343
[29]Sommer S,Fuqua S A W.Estrogen receptor and breast cancer[J].Semin Cancer Biol,2001,11(5):339-352
[30]Stoner M,Saville B,Wormke M,et al.Hypoxia induces proteasome dependent degradation of estrogen receptor alpha in ZR275 breast cancer cells[J].Mol Endocrinol, 2002,16 (10) :2231-2242
[31] Roger P,Daures J P,Maudelonde T, et al. Dissociated overexp ression of cathep sin D and estrogen recep tor alpha in p reinvasive mammary tumors [J]. Hum Pathol, 2000, 31 (5) : 593-600
[32] Donaghue C, Westley B R, May F E. Selective p romoter usage of the human estrogen receptor-alpha gene and its regulation by estrogen[J]. Mol Endocrinol, 1999, 13 (11) :1934-1950
[33] Lapidus R G, Nass S J, Butash K A, et al. Mapping of ER gene CpG island methylation-specific polymerase chain reaction[J]. Cancer Res, 1998,58(12): 2515-2519
[34] Zhao C, Lam E W, Sunters A, et al . Expression of estrogen receptor beta isoforms in normal breast epithelial cells and breast cancer: regulation by methylation[J]. Oncogene, 2003,22(48): 7600-7606
[35] Nass S J, Ferguson A T, El Ashry D, et al. Expression of DNA methyltransferase (DMT) and the cell cycle in human breast cancer cells [J]. Oncogene, 1999,18(52): 7453-7461
[36] Yang X, Ferguson A T, Nass S J, et al. Transcriptional activation of estrogen receptor alpha in human breast cancer cells by histone deacety- lase inhibition[J]. Cancer Res, 2000,60(24):6890-6894
[37] Yang X, Phillips D L, Ferguson A T, et al. Synergistic activation of functional estrogen receptor (ER)-alpha by DNA methyltransferase and histone deacetylase inhibition in human ER-alpha-negative breast cancer cells[J]. Cancer Res, 2001, 61(19): 7025-7029
[38] Zhu W G, Lakshmanan R R, Beal M D, et al. DNA methyltransferase inhibition enhances apoptosis induced by histone deacetylase inhibitors [J]. Cancer Res, 2001, 61(4): 1327-1333
[39] Billard L M, Magdinier F, Lenoir G M, et al. MeCP2 and MBD2 expression during normal and pathological growth of the human mammary gland [J]. Oncogene, 2002,21(17): 2704-2712
[40] Macaluso M, Cinti C, Russo G, et al. pRb2/ p130-E2F4/ 5-HDAC1- SUV39H1-p300 and pRb2/ p130-E2F4/ 5-HDAC1-SUV39H1-DNMT1 multimolecular complexes mediate the transcription of estrogen receptor alpha in breast cancer[J]. Oncogene, 2003, 22(23): 3511-3517
[41]Castagnetta L A,Miceli M D,Sorci C M,et al.Growth of LNCaP human prostate cancer cells is stimulated by estradiol via its own receptor[J].Endocrinology,1995,136(5):2309-2319
[42]朱英坚,叶敏.雌激素受体、雄激素受体与前列腺癌[J].国外医学泌尿系统分册,2002,22(5):283-286
[43]Li LC,Chui R,Nakajima K,et al.Frequent methylation of estrogen receptor in prostate cancer:Correlation with tumor progression[J].Cancer Res,2000,60(3):702-706
[44]Belinsky S A,Nikula K J,Baylin S B,et al.Increased cytosine DNA-methyl-transferase activity is target-cell-specific and an early event in lung cancer[j].Proc Natl Acad Sci USA,1996,93(9):4045-4050
[45]Lau K M,LaSpina M,Long J,et al.Expression of estrogen receptor(ER)-αand ER-β in normal and malignant prostatic epithelial cells:Regulation by methylation and involvement in growth regulation[J].Cancer Res,2000,60(12):3175-3182
[46]Schiff R,Massarweh S,Shou J,et al.Breast cancer endocrine resistance:how growth factor signaling and estrogen receptor coregulators modulate response[J].Clin Cancer Res,2003,9(1pt2):447S-454S
[47]Shiozawa T,Itoh K,Horiuchi A,et al.Down-regulation of estrogen receptor by the methylation of the estrogen receptor gene in endometrial carcinoma[J].Anticancer Res,2002,22(1A):139-143
[48]Hori M,Iwasaki M,Shimazaki J,et al.Assessment of hypermethylated DNA in two promoter regions of the estrogen receptor alpha gene in human endometrial diseases[J].Gynecol Oncol,2000,76(1):89-96
[49]Sasaki M,Kotcherguina L,Dharia A,et al.Cytosine- phosphoguanine methylation of estrogen receptors in endometrial cancer[J].Cancer Res,2001,61(8):3262-3266
[50]Ghabreau L,Roux J P,Niveleau A,et al.Correlation between the DNA global methylation status and progesterone receptor expression in normal endometrium,endometrioid adenocarcinoma and precursors[J].Virchows Arch,2004,445(2):129-134
[51]Fiegl H,Gattringer C,Widschwendter A,et al.Methylated DNA collected by tampons-a new tool to detect endometrial cancer[J].Cancer Epidemiol Biomarkers Prev,2004,13(5):882-888
[52]Brueckner B,Lyko F.DNA methyltransferase inhibitors:old and new drugs for an epigenetic cancer therapy[J].Trends Pharmacol Sci,2004,25(11):551-554
[53]Issa J P,Garcia-Manero G,Giles F J.Phase 1 study of low-dose prolonged exposure schedules of the hypomethylating agent 5-aza-2'- deoxycytidine(decitabine) in hematopoietic malignancies[J].Blood,2004,103(5):1635-1640
[54]Grodstein F,Martinez M E,Platz E A.Postmenopausal hormone use and risk for colonrectal cancer[J].Annals Internal Medicine,1998,128(9):705-712
[55]Nanda K,Bastian L A,Hasselblad V,et al.Hormone replacement therapy and the risk of colonrectal cancer:a meta-analysis.Obstet Gynecol,1999,93(5 SUPPL):880-888
[56]Issa J P,Ottaviano Y L,Celano P,et al.Methylation of the estrogen receptor CpG island links aging and neoplasia in human colon[J].Nature Genet,1994,7(4):536-540
[57]苏琦,敖启林,贺修胜,等.胃癌ER基因表达与CpG岛甲基化的意义[J].中国肿瘤临床,2004,31(11):607-610
[58]于力.DNA甲基化与肿瘤[J].中华内科杂志,2005,44(6):403-404