肾上腺髓质素在卵巢癌中的表达及其对ERK活性的影响
摘要
前言
卵巢癌是女性生殖器官常见的恶性肿瘤之一,发病率仅次于宫颈癌和子宫体癌而居第三位,但死亡率却居各类妇科肿瘤的首位。其特点是发现晚,进展快,治疗效果不理想,死亡率高,由于缺乏有效的早期诊断和治疗方法,发现时多属晚期,手术及化疗效果不佳。卵巢癌的发病机制十分复杂,现代分子生物学研究表明:恶性肿瘤的发生、发展不仅与肿瘤细胞的异常增殖、分化及细胞凋亡抑制有关,而且与信号转导机制异常有关。肾上腺髓质素(adrenomedullin,AM)是Kitamura等首次从人的肾上腺嗜铬细胞瘤中发现并分离出来的一种多功能肽类物质,由于在肾上腺髓质中的表达最高,所以被称为AM。现已知,AM主要由血管内皮细胞和血管平滑肌细胞合成分泌,随着对AM分子结构、生理作用以及信号转导机制研究的不断深入,发现AM在人类肿瘤的发生、侵袭及转移机制中也发挥重要的作用。现已知,有丝分裂原激活蛋白激酶(MAPK)/细胞外信号调节激酶(ERK)信号转导通路是AM的重要传导通路之一,是细胞信号转导的重要途径,ERK与恶性肿瘤预后密切相关。
本实验采用RT-PCR、MTT比色法及免疫印迹法检测卵巢上皮性恶性肿瘤,卵巢良性肿瘤及正常卵巢组织中,AM的表达情况及AM、AM受体阻断剂(AM22-52)对卵巢癌细胞增殖及ERK活性的影响,探讨AM在卵巢癌发生、发展中的作用及其信号转导途径,为卵巢癌的生物治疗提供新的实验依据。
材料与方法
一、临床资料
本实验标本取自2005年05月至2008年07月在中国医科大学附属第一医院住院行手术治疗,术前未行化疗和放疗,术后均经病理检查确诊的病例。组织学分类包括:正常卵巢组织10例(均来自年龄45-50岁,未绝经、患子宫肌瘤行预防性卵巢切除的患者)、卵巢上皮性良性肿瘤10例(浆液性囊腺瘤4例、粘液性囊腺瘤6例)、卵巢上皮性恶性肿瘤30例(浆液性囊腺癌21例、粘液性囊腺癌9例),共50例。卵巢上皮性恶性肿瘤病例分期标准参照国际妇产科联盟(FederationInternational of Gynecology and Obstetrics,FIGO)1986年手术病理分期标准:Ⅰ期10例、Ⅱ期10例、Ⅲ期10例。组织学分级,高分化(G1)17例,中分化(G2)5例,低分化(G3)8例。
二、实验方法
通过RT-PCR方法检测卵巢上皮性恶性肿瘤、卵巢上皮性良性肿瘤及正常卵巢组织中AM的表达量;体外培养卵巢癌CAVO3细胞株,加入不同浓度的AM及AM受体阻断剂(AM22-52),作用一定时间后MTT比色法检测细胞增殖情况;收集细胞,免疫印迹法检测ERK蛋白活性的变化。
三、统计学分析
实验数据以(?)±s表示,采用SPSS12.0统计软件进行独立样本t检验及相关分析,P＜0.05表示差异有统计学意义。
结果
(一) AM在卵巢上皮性恶性肿瘤、卵巢上皮性良性肿瘤及正常卵巢组织中的表达。
RT-PCR实验结果显示:AM在卵巢上皮性恶性肿瘤、卵巢上皮性良性肿瘤及正常卵巢组织中均有表达。在正常卵巢组织、卵巢上皮性良性肿瘤中呈低表达,在卵巢上皮性恶性肿瘤中呈高表达。其表达量与术后病理分级相关(P＜0.05),与手术病理分期无关。从而从基因表达水平验证了AM在上皮性卵巢癌中的表达情况。
(二)不同浓度的AM及AM受体阻断剂(AM22-52)在卵巢癌CAVO3细胞株中作用不同时间,检测CAVO3细胞增殖情况。
MTT比色法结果显示:AM促进卵巢癌CAVO3细胞株的增殖,AM四种不同浓度均表现为促进肿瘤增殖的作用,并且有明显的剂量依赖关系,随着作用时间的延长这种促进作用也随之增强,存在时间依赖关系,因此可以看出AM是CAOV3细胞增殖的促进剂。其两者之间存在剂量-时间依赖关系;同时实验中还观察到:AM受体阻断剂(AM22-52)抑制作用也存在剂量-时间的依赖关系,其作用与AM相反。从而从细胞水平验证AM在上皮性卵巢恶性肿瘤发生、发展中的作用。
(三)在加入不同浓度AM及AM受体阻断剂(AM22-52)的卵巢癌细胞株CAVO3中,检测ERK活性的变化。
免疫印迹法结果显示:AM可使ERK1/2活性升高。4种不同浓度的AM激动剂及AM受体阻断剂(AM22-52)(1×10~(-6)mol/L、1×10~(-7)mol/L、1×10~(-8)mol/L、1×10~(-9)mol/L)分别加入卵巢癌细胞株CAOV3中,用磷酸化的ERK1/2抗体检测ERK活性的变化。随着AM作用浓度的增加,磷酸化的ERK1/2的表达随之增高,反映ERK1/2活性随之升高;反之,随着AM受体阻断剂(AM22-52)浓度的增加,磷酸化的ERK1/2的表达随之降低,反映ERK1/2活性随之降低,两者之间存在着明显的剂量依赖关系,两者之间有统计学意义(P＜0.05)。
结论
1、AM在卵巢上皮性恶性肿瘤组织中呈现高表达,与术后病理分级密切相关。
2、AM能显著促进卵巢癌CAOV3细胞的增殖,AM受体阻断剂(AM22-52)则明显抑制卵巢癌CAOV3细胞的增殖,其两者之间存在着量效-时效的依赖关系。
3、AM能显著促进ERK活性,AM受体阻断剂(AM 22-52)则明显抑制ERK活性,两者之间存在着明显的剂量依赖关系。
Objective
Ovary cancer is one of common female malignant tumor.The disease incidence rate occupies the third place which is only inferior in cervical cancer and the uterus cancer.But the mortality rate occupies the first place in each kind of gynecology tumor.Its characteristic is discovers late,expands quickly,the treatment result is not ideal,the mortality rate is high.Because lacks of the effective early diagnosis and the method of treatment,When discovery is the later period,the surgery and the chemotherapy effect is not good.Pathogenesis of ovarian cancer is very complex.The modern molecular biology research indicated:carcinogenesis,development of malignant tumor not noly related with unusual multiplication of the differentiation of tumour cell and the cell perish weakly suppress,but also related with signal transduction mechanism exceptionally.The adrenomedullin is a kind of multi-purpose peptide class material which discovers from person's adrenal gland,Because of expression of adrenomedullin is highest,so called AM.Presently known,AM mainly synthesizes the secretion by the blood vessel endothelial cell and the blood vessel smooth muscle cell.Along with unceasingly thorough research AM molecular structure,physiological action as well as signal transduction mechanism.Discovered that it plays the vital role in the carcinogenesis,the attack and the shift mechanism of human tumor in AM.Presently known,the mitogen activated prote in kinase(MAPK)/extracellular signal regulated kinase(ERK) the signal transduction circuit is one of important conduction circuits in AM.it is the important way to cell signal transduction.ERK is a important link.ERK is closely related with the prognosis of malignant tumor.
The expression level of AM and receptor antagonist(AM22-52) evaluated by RT-PCR、MTT and western blot in ovarian cancer、normal ovarian and benign ovarian tumor.Discusses AM in the ovary carcinogenesis,development and the signal transduction pathway,provides the new experiment basis for biological treatment of ovarian cancer.
Methods
1、Patients
In this study,the samples were chosen from normal、benign and malignant ovarian cancer 50 cases who suffered from hysterectomy at the Dpartment of Gynecology,the First hospital of Medical University,from 2005-05 to 2008-07.They had been diagnosised correctly by routine pathological examination.None of the patients had received radiotherapy,chemotherapy,and other treatment before operation.normal ovarian tissue 10 cases,benign ovarian epithelial tumor 10 cases(serous cystadenoma 4 cases,mucinous cystadenoma 6 cases),ovarian malignant epithelial tumors 30 cases(serous cystadenocarcinoma 21 cases,Mucinous Cystadenocarcinoma 9 cases),total 50 cases,surgical-pathologic staging:stageⅠ10 cases,stageⅡ10 cases,stageⅢ10 cases,histological grade:well differentiated(G1) 17 cases,middle differentiation(G2)5 cases,poorly differentiated(G3) 8 cases.
2、Methods
The expression of AM in tissues from normal ovarian tissue、benign ovarian epithelial tumor、ovarian malignant epithelial tumors were detected by using RT-PCR method.Raise ovarian cancer CAVO3 cell line,Joins the different density AM and the AM acceptor blocking agent(AM22-52).After function certain time,MTT method examination cell multiplication situation.Collecting cell,western blot examines the ERK protein active change.
3、Statistical Analysis
Spss 12.0 software was employed to analyze all data.Independent-samples t test,P＜0.05 was considered as statistical significance。
Results
1、Expression of AM in normal ovarian tissue、benign ovarian epithelial tumor、ovarian malignant epithelial tumors.
The results of RT-PCR expressed of AM in normal ovarian tissue、benign ovarian epithelial tumor、ovarian malignant epithelial tumors.AM were expressed highly in the ovarian malignant epithelial tumors.but shows little expression in normal ovarian tissue and benign ovarian epithelial tumor.There was significantly difference between different histological grade,the expression of AM has no significant difference between clinical stage(P＜0.05).
2、Different density AM and AM acceptor blocking agent (AM22-52),affects the different time in the ovarian cancer CAVO3 cell line,examines the CAVO3 cell multiplication situation.
MTT result demonstration:AM promotion ovarian cancer multiplication in CAVO3 cell line.The AM four kind of different densities display to promote the function which the tumor multiplies.And has the obvious dosage dependence.Along with response time extension this kind of promoter action also along with it enhancement,it has existence time dependence.Therefore AM can promot the multiplication of CAOV3 cell.Between its both has the dosage-time dependence.The AM acceptor blocking agent(AM22-52) the inhibitory action also has the dosage-time the dependence.
3、After ovarian cancer cell line CAVO3 joins different density AM and the AM acceptor blocking agent(AM22-52),ERK protein active change.
Increases the ERK1/2 activeness along with the AM density to elevate along with it.Otherwise the density increases the ERK1/2 activeness along with the AM acceptor blocking agent(AM22-52) to reduce along with it.Between both has the obvious dosage dependence.Assumes with the AM density is remarkable is being related(AM22-52) assumes the remarkable inverse correlation with the AM acceptor blocking agent.
Conclusion
1、AM were expressed highly in the ovarian malignant epithelial tumors.There was significantly difference between different histological grade.
2、AM can obviously promote the ovarian cancer multiplication of CAOV3 cell, the AM acceptor blocking agent(AM22-52) obviously to suppress the ovarian cancer multiplication of CAOV3 cell.
3、AM can obviously promote the ERK activeness,the AM acceptor blocking agent(AM22-52) obviously to suppress ERK activeness.Between both has the obvious dosage dependence.
卵巢癌是女性生殖器官常见的恶性肿瘤之一,发病率仅次于宫颈癌和子宫体癌而居第三位,但死亡率却居各类妇科肿瘤的首位。其特点是发现晚,进展快,治疗效果不理想,死亡率高,由于缺乏有效的早期诊断和治疗方法,发现时多属晚期,手术及化疗效果不佳。卵巢癌的发病机制十分复杂,现代分子生物学研究表明:恶性肿瘤的发生、发展不仅与肿瘤细胞的异常增殖、分化及细胞凋亡抑制有关,而且与信号转导机制异常有关。肾上腺髓质素(adrenomedullin,AM)是Kitamura等首次从人的肾上腺嗜铬细胞瘤中发现并分离出来的一种多功能肽类物质,由于在肾上腺髓质中的表达最高,所以被称为AM。现已知,AM主要由血管内皮细胞和血管平滑肌细胞合成分泌,随着对AM分子结构、生理作用以及信号转导机制研究的不断深入,发现AM在人类肿瘤的发生、侵袭及转移机制中也发挥重要的作用。现已知,有丝分裂原激活蛋白激酶(MAPK)/细胞外信号调节激酶(ERK)信号转导通路是AM的重要传导通路之一,是细胞信号转导的重要途径,ERK与恶性肿瘤预后密切相关。
本实验采用RT-PCR、MTT比色法及免疫印迹法检测卵巢上皮性恶性肿瘤,卵巢良性肿瘤及正常卵巢组织中,AM的表达情况及AM、AM受体阻断剂(AM22-52)对卵巢癌细胞增殖及ERK活性的影响,探讨AM在卵巢癌发生、发展中的作用及其信号转导途径,为卵巢癌的生物治疗提供新的实验依据。
材料与方法
一、临床资料
本实验标本取自2005年05月至2008年07月在中国医科大学附属第一医院住院行手术治疗,术前未行化疗和放疗,术后均经病理检查确诊的病例。组织学分类包括:正常卵巢组织10例(均来自年龄45-50岁,未绝经、患子宫肌瘤行预防性卵巢切除的患者)、卵巢上皮性良性肿瘤10例(浆液性囊腺瘤4例、粘液性囊腺瘤6例)、卵巢上皮性恶性肿瘤30例(浆液性囊腺癌21例、粘液性囊腺癌9例),共50例。卵巢上皮性恶性肿瘤病例分期标准参照国际妇产科联盟(FederationInternational of Gynecology and Obstetrics,FIGO)1986年手术病理分期标准:Ⅰ期10例、Ⅱ期10例、Ⅲ期10例。组织学分级,高分化(G1)17例,中分化(G2)5例,低分化(G3)8例。
二、实验方法
通过RT-PCR方法检测卵巢上皮性恶性肿瘤、卵巢上皮性良性肿瘤及正常卵巢组织中AM的表达量;体外培养卵巢癌CAVO3细胞株,加入不同浓度的AM及AM受体阻断剂(AM22-52),作用一定时间后MTT比色法检测细胞增殖情况;收集细胞,免疫印迹法检测ERK蛋白活性的变化。
三、统计学分析
实验数据以(?)±s表示,采用SPSS12.0统计软件进行独立样本t检验及相关分析,P＜0.05表示差异有统计学意义。
结果
(一) AM在卵巢上皮性恶性肿瘤、卵巢上皮性良性肿瘤及正常卵巢组织中的表达。
RT-PCR实验结果显示:AM在卵巢上皮性恶性肿瘤、卵巢上皮性良性肿瘤及正常卵巢组织中均有表达。在正常卵巢组织、卵巢上皮性良性肿瘤中呈低表达,在卵巢上皮性恶性肿瘤中呈高表达。其表达量与术后病理分级相关(P＜0.05),与手术病理分期无关。从而从基因表达水平验证了AM在上皮性卵巢癌中的表达情况。
(二)不同浓度的AM及AM受体阻断剂(AM22-52)在卵巢癌CAVO3细胞株中作用不同时间,检测CAVO3细胞增殖情况。
MTT比色法结果显示:AM促进卵巢癌CAVO3细胞株的增殖,AM四种不同浓度均表现为促进肿瘤增殖的作用,并且有明显的剂量依赖关系,随着作用时间的延长这种促进作用也随之增强,存在时间依赖关系,因此可以看出AM是CAOV3细胞增殖的促进剂。其两者之间存在剂量-时间依赖关系;同时实验中还观察到:AM受体阻断剂(AM22-52)抑制作用也存在剂量-时间的依赖关系,其作用与AM相反。从而从细胞水平验证AM在上皮性卵巢恶性肿瘤发生、发展中的作用。
(三)在加入不同浓度AM及AM受体阻断剂(AM22-52)的卵巢癌细胞株CAVO3中,检测ERK活性的变化。
免疫印迹法结果显示:AM可使ERK1/2活性升高。4种不同浓度的AM激动剂及AM受体阻断剂(AM22-52)(1×10~(-6)mol/L、1×10~(-7)mol/L、1×10~(-8)mol/L、1×10~(-9)mol/L)分别加入卵巢癌细胞株CAOV3中,用磷酸化的ERK1/2抗体检测ERK活性的变化。随着AM作用浓度的增加,磷酸化的ERK1/2的表达随之增高,反映ERK1/2活性随之升高;反之,随着AM受体阻断剂(AM22-52)浓度的增加,磷酸化的ERK1/2的表达随之降低,反映ERK1/2活性随之降低,两者之间存在着明显的剂量依赖关系,两者之间有统计学意义(P＜0.05)。
结论
1、AM在卵巢上皮性恶性肿瘤组织中呈现高表达,与术后病理分级密切相关。
2、AM能显著促进卵巢癌CAOV3细胞的增殖,AM受体阻断剂(AM22-52)则明显抑制卵巢癌CAOV3细胞的增殖,其两者之间存在着量效-时效的依赖关系。
3、AM能显著促进ERK活性,AM受体阻断剂(AM 22-52)则明显抑制ERK活性,两者之间存在着明显的剂量依赖关系。
Objective
Ovary cancer is one of common female malignant tumor.The disease incidence rate occupies the third place which is only inferior in cervical cancer and the uterus cancer.But the mortality rate occupies the first place in each kind of gynecology tumor.Its characteristic is discovers late,expands quickly,the treatment result is not ideal,the mortality rate is high.Because lacks of the effective early diagnosis and the method of treatment,When discovery is the later period,the surgery and the chemotherapy effect is not good.Pathogenesis of ovarian cancer is very complex.The modern molecular biology research indicated:carcinogenesis,development of malignant tumor not noly related with unusual multiplication of the differentiation of tumour cell and the cell perish weakly suppress,but also related with signal transduction mechanism exceptionally.The adrenomedullin is a kind of multi-purpose peptide class material which discovers from person's adrenal gland,Because of expression of adrenomedullin is highest,so called AM.Presently known,AM mainly synthesizes the secretion by the blood vessel endothelial cell and the blood vessel smooth muscle cell.Along with unceasingly thorough research AM molecular structure,physiological action as well as signal transduction mechanism.Discovered that it plays the vital role in the carcinogenesis,the attack and the shift mechanism of human tumor in AM.Presently known,the mitogen activated prote in kinase(MAPK)/extracellular signal regulated kinase(ERK) the signal transduction circuit is one of important conduction circuits in AM.it is the important way to cell signal transduction.ERK is a important link.ERK is closely related with the prognosis of malignant tumor.
The expression level of AM and receptor antagonist(AM22-52) evaluated by RT-PCR、MTT and western blot in ovarian cancer、normal ovarian and benign ovarian tumor.Discusses AM in the ovary carcinogenesis,development and the signal transduction pathway,provides the new experiment basis for biological treatment of ovarian cancer.
Methods
1、Patients
In this study,the samples were chosen from normal、benign and malignant ovarian cancer 50 cases who suffered from hysterectomy at the Dpartment of Gynecology,the First hospital of Medical University,from 2005-05 to 2008-07.They had been diagnosised correctly by routine pathological examination.None of the patients had received radiotherapy,chemotherapy,and other treatment before operation.normal ovarian tissue 10 cases,benign ovarian epithelial tumor 10 cases(serous cystadenoma 4 cases,mucinous cystadenoma 6 cases),ovarian malignant epithelial tumors 30 cases(serous cystadenocarcinoma 21 cases,Mucinous Cystadenocarcinoma 9 cases),total 50 cases,surgical-pathologic staging:stageⅠ10 cases,stageⅡ10 cases,stageⅢ10 cases,histological grade:well differentiated(G1) 17 cases,middle differentiation(G2)5 cases,poorly differentiated(G3) 8 cases.
2、Methods
The expression of AM in tissues from normal ovarian tissue、benign ovarian epithelial tumor、ovarian malignant epithelial tumors were detected by using RT-PCR method.Raise ovarian cancer CAVO3 cell line,Joins the different density AM and the AM acceptor blocking agent(AM22-52).After function certain time,MTT method examination cell multiplication situation.Collecting cell,western blot examines the ERK protein active change.
3、Statistical Analysis
Spss 12.0 software was employed to analyze all data.Independent-samples t test,P＜0.05 was considered as statistical significance。
Results
1、Expression of AM in normal ovarian tissue、benign ovarian epithelial tumor、ovarian malignant epithelial tumors.
The results of RT-PCR expressed of AM in normal ovarian tissue、benign ovarian epithelial tumor、ovarian malignant epithelial tumors.AM were expressed highly in the ovarian malignant epithelial tumors.but shows little expression in normal ovarian tissue and benign ovarian epithelial tumor.There was significantly difference between different histological grade,the expression of AM has no significant difference between clinical stage(P＜0.05).
2、Different density AM and AM acceptor blocking agent (AM22-52),affects the different time in the ovarian cancer CAVO3 cell line,examines the CAVO3 cell multiplication situation.
MTT result demonstration:AM promotion ovarian cancer multiplication in CAVO3 cell line.The AM four kind of different densities display to promote the function which the tumor multiplies.And has the obvious dosage dependence.Along with response time extension this kind of promoter action also along with it enhancement,it has existence time dependence.Therefore AM can promot the multiplication of CAOV3 cell.Between its both has the dosage-time dependence.The AM acceptor blocking agent(AM22-52) the inhibitory action also has the dosage-time the dependence.
3、After ovarian cancer cell line CAVO3 joins different density AM and the AM acceptor blocking agent(AM22-52),ERK protein active change.
Increases the ERK1/2 activeness along with the AM density to elevate along with it.Otherwise the density increases the ERK1/2 activeness along with the AM acceptor blocking agent(AM22-52) to reduce along with it.Between both has the obvious dosage dependence.Assumes with the AM density is remarkable is being related(AM22-52) assumes the remarkable inverse correlation with the AM acceptor blocking agent.
Conclusion
1、AM were expressed highly in the ovarian malignant epithelial tumors.There was significantly difference between different histological grade.
2、AM can obviously promote the ovarian cancer multiplication of CAOV3 cell, the AM acceptor blocking agent(AM22-52) obviously to suppress the ovarian cancer multiplication of CAOV3 cell.
3、AM can obviously promote the ERK activeness,the AM acceptor blocking agent(AM22-52) obviously to suppress ERK activeness.Between both has the obvious dosage dependence.
引文
1 Ataajan M,Saravanan SM,ElsonDL.Advanced ovarian carcinoma as a chronic disease:a case report and reriew.S D J Med.2003,56(12):515-521.
2 Allaker RP,Zihni C,Kapas S.An investigation into the antimicrobial effects of adrenomedullin on members of the skin,oral respiratory tract and gut microflora.FEMS Immunol Med Microbiol,1999,23:289-293.
3 Hay DL,Smith DM.Adrenomedullin receptors:molecular identity and function[J].Peptides,2001,22(11):1753-1763.
4 Juaneda C,Dumont Y,Quirion R.The molecular pharmacology of CGRP and related peptide receptor subtypes[J].Trends Pharmacol Sci,2000,21(11):432-438.
5 Choi KC,Auersperg N,Leung PC.Mitogen-activated protein kinases in normal and(pre)neoplastic ovarian surface epithelium.Reprod Biol Endocrinol.2003,7(1):71-78.
6 Rosemary SZ,Heathereather AW,Pingyu Z,et al.Mechanisms Regulating the Constitutive Activation of the Extrecellular Signal-Regulated Kinase(ERK) Signaling Pathway in Ovarian Cancer and the Effect of Ribonucleic Acid Interference for ERK1/2 on Cancer Cell Proliferation.Molecular Endocrinology.2004,18(10):2570-2582.
7 韦淑琴,隋丽华,戚基萍,等。卵巢上皮性肿瘤组织中ERK的表达与活化。哈尔滨医科大学学报,2004,38(3):233-236.
8 Hata K,Takebayashi Y,Akiba S,et al.Expression of the adrenomedullin gene in epithelial ovarian cancer.Mol Hum Reprod.2000 Oct;6(10):867-872.
9 Giacalone PL,Vuaroqueaux V,Daures JP,et al.Expression of adrenomedullin in human ovaries,ovarian cysts and cancers.Correlation with estrogens receptor status.Eur J Obstet Gynecol Reprod Biol.2003 Oct 10;110(2):224-229.
10 王越,姚智。IL-6、IL-8与卵巢癌。国外医学分册,2004,31(12);939-943.
11 郭科军,张颐.Genistein和PD98059对aFGF及bFGF诱导的CAOV3细胞增殖的抑制作用.中国医科大学学报,2004,10(5):450-453.
12 Oehler MK,Norbury C,Hague S,et al.Adrenomedullin inhibits hypoxic cell death by upregulation of Bcl-2 in endometrial cancer cells:a possible promotion mechanism for tumour growth.Oncogene,2001,20:2937-2945.
13 Satoh F,Takahashi K,Murakami O,et al.Adrenomedullin in human brain,adrenal glands andtumor tissues of pheochromocytoma ganglioneuro blastoma and neuroblastoma.J Clin Endocrinol Metab,1995,80:1750-1752.
14 Martinez A,Miller MJ,Unsworth EJ,et al.Expression of adenomedullin in normal humanl ung and in pulmonary tumors.Endocrinology,1995,136:4099-4105.
15 Martinez A,Vos M,Guedez L,et al.The effects of adrenomedullin overexpression in breast tumor cell. J Natt Cancer Inst,2002,94(16): 1226-1237.
16 Oehler MK,Hague S,Rees MC, et al. Adrenomedullin promotes formation of xenografted endometrial tumors by stimulation of autocrine growth and aogenesis. Oncogene, 2002, 21(18):2815-2821.
17 Ousfik L ,Sauze S, Boudouresque F, et al. Neutralization of adrenomedullin inhibits the growth of human glioblastoma cell lines in vitro and suppresses tumor xenograft growth in vivo. Am J Pathol,2002,160(4): 1279-1292.
18 Miyashita K, Itoh H, Sawada N, et al. Adrenomedullin promotes proliferation and migration of cultured endothelial cells. Hypertens Res,2003, 26: 93-98.
19 Frede S, Freitag P, Otto T,et al. The proinflammatory cytokine interleukin 1 beta and hypoxia cooperatively induce the expression of adrenomedullin in ovarian carcinoma cells through hypoxia inducible factor 1 activation (J) .Cancer Res,2005,65(l1) :4690.
20 Zudaire E,Martinez A,Garayoa M,et al.Adrenomedullinis a cross talk molecule that regulates tumor and mast cell function during human carcinogenesis [J] . Am J Pathol,2006,168(l) :280. 2006,7 (21 ) :373-375.
22 Oehler MK, Fischer DC, Orlowska-Volk M, et al. Tissue and plasma expression of the angiogenic peptide adrenomedullin in breast cancer (J) Br J Cancer, 2003, 89(10) ::1927.
23 Michishita M, Minegishi T, Abe K, et al. Expression of adrenomedullin in the endometrium of the human uterus. Obstet Gyneco,1999, 93: 66-70.
24 Li Z, Takeuchi S, Otani T, et al. Implications of adrenomedullin expression in the invasion of squamous cell carcinoma of the uterine cervix. Int J Clin Oncol,2001, 6: 263-270.
25 Moriyama T, Otani T, Maruo T. Expression of adrenomedullin by human placental cytotrophoblasts and choriocarcinoma JAr cells. J Clin Endocrinol Metab, 2001, 86 :3958-3961.
26 Herrera R. Modulation of hepatocyte growth factor - induced scattering of HT29 colon cancer cells. Involvement of the MAPK pathway[J]. J Cell Sci, 1998, 11 l(pt8):1039-1049.
1 Sakata J, Shimokubo T, Kitamura K, et al.Molecular cloning and biological activities of rat adrenomedullin, a hypotensive peptidel[J].Biochem Biophys Res Commun, 1993, 195: 9212-9271.
2 JougasakiM, Stevens TL, Borgeson DD, et al. Adrenomedullin in experimental congestive heart failure: cardiorenal activation[J].Am J Physiol, 1997, 273: 1392-1399.
3 Beltowski J ,Jamroz A. Adrenomedullin what do we know 10 years since its discovery? [J]. Pol J Pharmacol,2004 ,56 ,5227.
4 J uancda C ,Dumont Y,Quirion R. molecular pharmacology of CGRP and related peptide receptor subtypes[J].Trends Pharmacol Sci ,2000 ,21(11) :4322-4328.
5 Parameswaran N , Spielman WS , Brooks DP , et al.SB-03580 reverses adrenomedullin ' s effect on proliferation and apoptosis in cultured mesangial cells[J].Eur J Pharma,1999 ,371(1) :75-82.
6 Parameswaran N,Hall CS,McCabe LR ,et al.Adrenomedullin increases AP-1 expression in rat mesangial cells via activation of protein kinase-A and p38 MAPK[J]. Cell Physiol Biochem,2003,13(6):367-374.
7 Ouafik L, Sauze S, Boudouresque F, et al. Neutralization of adrenomedullin inhibits the growth of human glioblastoma cell lines in vitro and suppresses tumor xenograft growth in vivo[J]. Am J Pathol ,2002,160(4):1279-1292
8 Martinez A, Vos M, Guedez L, et al. The effects of adrenomedullin overexpression in breast tumor cell[J]. J Natt CancerInst ,2002,94:1226-1237.
9 Kamoi H, Kanazawa H, Hirata K, et al. Adrenomedullin inhibits the secretion of cytokine-induced neutrophil chemoattractant, a member of the interleukin-8 family, from rat alveolar macrophages[J]. Biochem Biophys Res Commun ,1995,211(3):1031-1035.
10 Iimuro S, Shindo T, Moriyama N, et al. Angiogenic effects of adrenomedullin in ischemia and tumor growth[J]. CircRes,2004,95(4):415-423.
11 OehlerMK, Norbury C, Hague S, et al. Adrenomedullin inhibits hypoxic cell death by up regulation of Bcl-2 in endometrial cancer cells: a possible promotion mechanism for tumour growth[J]. Oncogene,2001, 20: 2937-2945.
12 MichishitaM,Minegishi T, Abe K, et al. Expression of adrenomedullin in the endometriun of the human uterus[J].Obstet Gynecol,1999,93(1): 66-70.
13 @@@, 2007,30(4),231-234.
14 Li Z, Takeuchi S, Otani T, et al.Implications of adrenomedullin expression in the invasion of squamous cell carcinoma of the uterine cervixl[J].Int J Clin Oncol, 2001, 6:263-270.
15 Li Z, Takeuchi S, Ohara N, et al. Paradoxically abundant expression of Bcl-2 and adrenomedullin in invasive cervical squamous carcinoma[J].Int J Clin Oncol,2003,8(2):83-89.
16 Hata K,Takebayashi Y,Akiba S,et al.Expression of the adrenomedullin gene in epithelial ovarian cancer[J].Mol Hum Reprod,2002,6(10):867-872.
17 Moriyama T,Otani T,Maruo T.Expression of adrenomedullin by human placental cytotrophoblasts and choriocarcinoma JAr cells[J].J Clin EndocrinolMetab,2001,86:3958-3961.
18 江元,田雪红,袁捷,等.肾上腺髓质素在子宫平滑肌肿瘤组织中的表达及其与血管生成和预后的研究[J].中国实用妇科与产科杂志,2006,22(4):262-264.
19 Rice PI,Goldberg RJ,Ray EC,et al.Inhibition of extracellular signal - regulated kinase 1/2phosphorylation and induction of apotosis by sulindac metabolites[J].Cancer Res,2001,61(4):1541-1547.
20 Krueger JS,Song RX,Mcpherson R,et al.Temporal and quantitative regulation of mitogen activated protein kinase(MAPK)modulates cell motility and invasion[J].Oncogene,2001,20(31):4209- 4218.
21 赵杨,郭科军,张颐,等.细胞外信号调节激酶在卵巢癌中的表达及临床意义[J].中国肿瘤临床,2006,33(17):978-981.
22 Givant,Horwitz V,Davidson B,Lazarovici P,et al.Mitogen-activated protein kinases (MAPK) as predictors of clinical outcome in serous ovarian carcinoma in effusions[J].Gynecol Oncol,2003,91(1):160-172.
2 Allaker RP,Zihni C,Kapas S.An investigation into the antimicrobial effects of adrenomedullin on members of the skin,oral respiratory tract and gut microflora.FEMS Immunol Med Microbiol,1999,23:289-293.
3 Hay DL,Smith DM.Adrenomedullin receptors:molecular identity and function[J].Peptides,2001,22(11):1753-1763.
4 Juaneda C,Dumont Y,Quirion R.The molecular pharmacology of CGRP and related peptide receptor subtypes[J].Trends Pharmacol Sci,2000,21(11):432-438.
5 Choi KC,Auersperg N,Leung PC.Mitogen-activated protein kinases in normal and(pre)neoplastic ovarian surface epithelium.Reprod Biol Endocrinol.2003,7(1):71-78.
6 Rosemary SZ,Heathereather AW,Pingyu Z,et al.Mechanisms Regulating the Constitutive Activation of the Extrecellular Signal-Regulated Kinase(ERK) Signaling Pathway in Ovarian Cancer and the Effect of Ribonucleic Acid Interference for ERK1/2 on Cancer Cell Proliferation.Molecular Endocrinology.2004,18(10):2570-2582.
7 韦淑琴,隋丽华,戚基萍,等。卵巢上皮性肿瘤组织中ERK的表达与活化。哈尔滨医科大学学报,2004,38(3):233-236.
8 Hata K,Takebayashi Y,Akiba S,et al.Expression of the adrenomedullin gene in epithelial ovarian cancer.Mol Hum Reprod.2000 Oct;6(10):867-872.
9 Giacalone PL,Vuaroqueaux V,Daures JP,et al.Expression of adrenomedullin in human ovaries,ovarian cysts and cancers.Correlation with estrogens receptor status.Eur J Obstet Gynecol Reprod Biol.2003 Oct 10;110(2):224-229.
10 王越,姚智。IL-6、IL-8与卵巢癌。国外医学分册,2004,31(12);939-943.
11 郭科军,张颐.Genistein和PD98059对aFGF及bFGF诱导的CAOV3细胞增殖的抑制作用.中国医科大学学报,2004,10(5):450-453.
12 Oehler MK,Norbury C,Hague S,et al.Adrenomedullin inhibits hypoxic cell death by upregulation of Bcl-2 in endometrial cancer cells:a possible promotion mechanism for tumour growth.Oncogene,2001,20:2937-2945.
13 Satoh F,Takahashi K,Murakami O,et al.Adrenomedullin in human brain,adrenal glands andtumor tissues of pheochromocytoma ganglioneuro blastoma and neuroblastoma.J Clin Endocrinol Metab,1995,80:1750-1752.
14 Martinez A,Miller MJ,Unsworth EJ,et al.Expression of adenomedullin in normal humanl ung and in pulmonary tumors.Endocrinology,1995,136:4099-4105.
15 Martinez A,Vos M,Guedez L,et al.The effects of adrenomedullin overexpression in breast tumor cell. J Natt Cancer Inst,2002,94(16): 1226-1237.
16 Oehler MK,Hague S,Rees MC, et al. Adrenomedullin promotes formation of xenografted endometrial tumors by stimulation of autocrine growth and aogenesis. Oncogene, 2002, 21(18):2815-2821.
17 Ousfik L ,Sauze S, Boudouresque F, et al. Neutralization of adrenomedullin inhibits the growth of human glioblastoma cell lines in vitro and suppresses tumor xenograft growth in vivo. Am J Pathol,2002,160(4): 1279-1292.
18 Miyashita K, Itoh H, Sawada N, et al. Adrenomedullin promotes proliferation and migration of cultured endothelial cells. Hypertens Res,2003, 26: 93-98.
19 Frede S, Freitag P, Otto T,et al. The proinflammatory cytokine interleukin 1 beta and hypoxia cooperatively induce the expression of adrenomedullin in ovarian carcinoma cells through hypoxia inducible factor 1 activation (J) .Cancer Res,2005,65(l1) :4690.
20 Zudaire E,Martinez A,Garayoa M,et al.Adrenomedullinis a cross talk molecule that regulates tumor and mast cell function during human carcinogenesis [J] . Am J Pathol,2006,168(l) :280. 2006,7 (21 ) :373-375.
22 Oehler MK, Fischer DC, Orlowska-Volk M, et al. Tissue and plasma expression of the angiogenic peptide adrenomedullin in breast cancer (J) Br J Cancer, 2003, 89(10) ::1927.
23 Michishita M, Minegishi T, Abe K, et al. Expression of adrenomedullin in the endometrium of the human uterus. Obstet Gyneco,1999, 93: 66-70.
24 Li Z, Takeuchi S, Otani T, et al. Implications of adrenomedullin expression in the invasion of squamous cell carcinoma of the uterine cervix. Int J Clin Oncol,2001, 6: 263-270.
25 Moriyama T, Otani T, Maruo T. Expression of adrenomedullin by human placental cytotrophoblasts and choriocarcinoma JAr cells. J Clin Endocrinol Metab, 2001, 86 :3958-3961.
26 Herrera R. Modulation of hepatocyte growth factor - induced scattering of HT29 colon cancer cells. Involvement of the MAPK pathway[J]. J Cell Sci, 1998, 11 l(pt8):1039-1049.
1 Sakata J, Shimokubo T, Kitamura K, et al.Molecular cloning and biological activities of rat adrenomedullin, a hypotensive peptidel[J].Biochem Biophys Res Commun, 1993, 195: 9212-9271.
2 JougasakiM, Stevens TL, Borgeson DD, et al. Adrenomedullin in experimental congestive heart failure: cardiorenal activation[J].Am J Physiol, 1997, 273: 1392-1399.
3 Beltowski J ,Jamroz A. Adrenomedullin what do we know 10 years since its discovery? [J]. Pol J Pharmacol,2004 ,56 ,5227.
4 J uancda C ,Dumont Y,Quirion R. molecular pharmacology of CGRP and related peptide receptor subtypes[J].Trends Pharmacol Sci ,2000 ,21(11) :4322-4328.
5 Parameswaran N , Spielman WS , Brooks DP , et al.SB-03580 reverses adrenomedullin ' s effect on proliferation and apoptosis in cultured mesangial cells[J].Eur J Pharma,1999 ,371(1) :75-82.
6 Parameswaran N,Hall CS,McCabe LR ,et al.Adrenomedullin increases AP-1 expression in rat mesangial cells via activation of protein kinase-A and p38 MAPK[J]. Cell Physiol Biochem,2003,13(6):367-374.
7 Ouafik L, Sauze S, Boudouresque F, et al. Neutralization of adrenomedullin inhibits the growth of human glioblastoma cell lines in vitro and suppresses tumor xenograft growth in vivo[J]. Am J Pathol ,2002,160(4):1279-1292
8 Martinez A, Vos M, Guedez L, et al. The effects of adrenomedullin overexpression in breast tumor cell[J]. J Natt CancerInst ,2002,94:1226-1237.
9 Kamoi H, Kanazawa H, Hirata K, et al. Adrenomedullin inhibits the secretion of cytokine-induced neutrophil chemoattractant, a member of the interleukin-8 family, from rat alveolar macrophages[J]. Biochem Biophys Res Commun ,1995,211(3):1031-1035.
10 Iimuro S, Shindo T, Moriyama N, et al. Angiogenic effects of adrenomedullin in ischemia and tumor growth[J]. CircRes,2004,95(4):415-423.
11 OehlerMK, Norbury C, Hague S, et al. Adrenomedullin inhibits hypoxic cell death by up regulation of Bcl-2 in endometrial cancer cells: a possible promotion mechanism for tumour growth[J]. Oncogene,2001, 20: 2937-2945.
12 MichishitaM,Minegishi T, Abe K, et al. Expression of adrenomedullin in the endometriun of the human uterus[J].Obstet Gynecol,1999,93(1): 66-70.
13 @@@, 2007,30(4),231-234.
14 Li Z, Takeuchi S, Otani T, et al.Implications of adrenomedullin expression in the invasion of squamous cell carcinoma of the uterine cervixl[J].Int J Clin Oncol, 2001, 6:263-270.
15 Li Z, Takeuchi S, Ohara N, et al. Paradoxically abundant expression of Bcl-2 and adrenomedullin in invasive cervical squamous carcinoma[J].Int J Clin Oncol,2003,8(2):83-89.
16 Hata K,Takebayashi Y,Akiba S,et al.Expression of the adrenomedullin gene in epithelial ovarian cancer[J].Mol Hum Reprod,2002,6(10):867-872.
17 Moriyama T,Otani T,Maruo T.Expression of adrenomedullin by human placental cytotrophoblasts and choriocarcinoma JAr cells[J].J Clin EndocrinolMetab,2001,86:3958-3961.
18 江元,田雪红,袁捷,等.肾上腺髓质素在子宫平滑肌肿瘤组织中的表达及其与血管生成和预后的研究[J].中国实用妇科与产科杂志,2006,22(4):262-264.
19 Rice PI,Goldberg RJ,Ray EC,et al.Inhibition of extracellular signal - regulated kinase 1/2phosphorylation and induction of apotosis by sulindac metabolites[J].Cancer Res,2001,61(4):1541-1547.
20 Krueger JS,Song RX,Mcpherson R,et al.Temporal and quantitative regulation of mitogen activated protein kinase(MAPK)modulates cell motility and invasion[J].Oncogene,2001,20(31):4209- 4218.
21 赵杨,郭科军,张颐,等.细胞外信号调节激酶在卵巢癌中的表达及临床意义[J].中国肿瘤临床,2006,33(17):978-981.
22 Givant,Horwitz V,Davidson B,Lazarovici P,et al.Mitogen-activated protein kinases (MAPK) as predictors of clinical outcome in serous ovarian carcinoma in effusions[J].Gynecol Oncol,2003,91(1):160-172.