沙利度胺联合表阿霉素治疗小鼠H22肝癌移植瘤的实验研究及其机制的探讨
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摘要
目的:通过构建小鼠H22肝癌移植瘤模型,观察沙利度胺联合表阿霉素对小鼠H22肝癌移植瘤生长的影响,并初步探讨沙利度胺抗肿瘤的作用机制。
方法:取40只健康雄性昆明种小鼠,无菌条件下将小鼠H22肝癌腹水瘤细胞接种到小鼠右腋皮下,0.2ml/只(2.0×106个细胞),构建荷瘤肝癌小鼠模型。接种第二天将荷瘤小鼠随机分为4组,每组10只:沙利度胺组(灌胃沙利度胺200mg/kg/日,连续给药10天)、沙利度胺联合表阿霉素组(灌胃沙利度胺200mg/kg/日,连续给药10天;腹腔注射表阿霉素50mg/m2,接种第二天给药一次)、表阿霉素组(腹腔注射表阿霉素50mg/m2,接种第二天给药一次)、生理盐水对照组(灌胃生理盐水0.3ml/日,连续给药10天),停药次日脱臼处死,完整剥离瘤组织,称重,计算抑瘤率及两药相互作用系数。分别提取各组瘤细胞总RNA,鉴定其完整性及含量,采用半定量RT-PCR方法检测肿瘤组织内bFGF mRNA、MMP-2 mRNA的表达水平,应用免疫组化方法检测肿瘤组织内bFGF、MMP-2蛋白的表达水平。用SPSS15.0统计学软件进行数据处理,各组数据以均值±标准差的形式表示。
结果:
1沙利度胺组瘤重为(3.67±0.44)g,抑瘤率为19.52%,沙利度胺联合表阿霉素组瘤重为(1.95±0.24)g,抑瘤率为57.24%,表阿霉素组瘤重为(2.45±0.31)g,抑瘤率为46.27%,均与生理盐水对照组相比有统计学意义(P<0.01)。生理盐水对照组瘤重为(4.56±0.46)g。两药相互作用系数(CDI)=0.9889<1,沙利度胺联合表阿霉素组平均瘤重最低,与沙利度胺组、表阿霉素组相比均有统计学意义(P<0.01)。
2采用半定量RT-PCR的方法检测各组肿瘤组织内bFGF mRNA、MMP-2 mRNA的表达水平。沙利度胺组bFGF mRNA表达相对值为0.3307±0.0463,沙利度胺联合表阿霉素组bFGF mRNA表达相对值为0.3054±0.0539,均与生理盐水对照组相比有统计学差异(P<0.01)。生理盐水对照组bFGF mRNA表达相对值为0.6732±0.1244,表阿霉素组bFGF mRNA表达相对值为0.6113±0.0629。生理盐水对照组和表阿霉素组相比无统计学意义(P>0.05)。沙利度胺组MMP-2 mRNA表达相对值为0.2835±0.0569,沙利度胺联合表阿霉素组MMP-2 mRNA表达相对值为0.2502±0.069,均与生理盐水对照组相比有统计学意义(P<0.01)。生理盐水对照组MMP-2 mRNA表达相对值为0.716±0.1126,表阿霉素组MMP-2 mRNA表达相对值为0.6847±0.1002,生理盐水对照组和表阿霉素组相比无统计学意义(P>0.05)。
3应用免疫组化方法检测各组肿瘤组织内bFGF、MMP-2蛋白的表达水平。沙利度胺组bFGF表达值为2.4±0.9661,沙利度胺联合表阿霉素组bFGF表达值为2.3±0.675,均与生理盐水对照组相比有统计学意义(P<0.01)。生理盐水对照组bFGF表达值为5.2±0.7888,表阿霉素组bFGF表达值为4.5±1.1785,表阿霉素组和生理盐水对照组相比无统计学意义(P>0.05)。沙利度胺组MMP-2表达值为2.0±0.6667,沙利度胺联合表阿霉素组MMP-2表达值为1.9±0.7379,均与生理盐水对照组相比有统计学意义(P<0.01)。生理盐水对照组MMP-2表达值为3.8±1.3166,表阿霉素组MMP-2表达值为3.2±1.0328,表阿霉素组和生理盐水对照组相比无统计学意义(P>0.05)。
结论:
1沙利度胺具有抑制小鼠H22肝癌移植瘤增殖的作用。
2沙利度胺可协同表阿霉素对小鼠H22肝癌移植瘤增殖的抑制作用。
3沙利度胺可能通过下调bFGF、MMP-2的表达而发挥抑制小鼠H22肝癌移植瘤增殖作用。
4本实验证实了沙利度胺具有抗肝癌作用,并通过沙利度胺联合抗肿瘤药物表阿霉素的实验研究丰富了肝癌的治疗,为肝癌的综合治疗提供了新的选择。
Objective: Established an H22 mice hepatocellular carcinoma model, the effict of Thalidomide(Tha) combined with Epirubicine(Epi) on H22 murine hepatocellular carcinoma was observed, the aim was to explore the possible mechanism of thalidomide anti-tumor.
Methods: Fourty healthy male kunming mice were transplanted by right armpit injection of H22 tumor cells in asepsis conditions, 0.2ml per mice(2.0×106 cells). Then these mice were randomly divided into four groups, ten mice in each group. Tha group (oral Tha 200mg/kg/d, d1-10), Tha+Epi group (oral Tha 200mg/kg/d, d1-10; Epirubicine intraperitoneal injection, 50mg/m2, d1), Epi group (Epirubicine intraperitoneal injection,50mg/m2, d1), Normal Saline group (oral normal saline 0.3ml/d, d1-10). Ten days later all mice were killed and the tumor were taken and weighed. Then the ratio of tumor suppression could be calculated, coefficient of drug interaction(CDI) was calculated. The expression of bFGF mRNA and MMP-2 mRNA were examined by reverse transcription polym-erase chain reaction(RT-PCR). The expression of bFGF, MMP-2 in tumor tissue was detected by using immunohistochemical (IHC) method. All data was showed by Mean±SD. The data was analyzed with the SPSS15.0 software.
Results:
1 The tumor weight of Tha group was (3.67±0.44)g, the inhibition rate was 19.52%; The tumor weight of Tha+Epi group was (1.95±0.24)g, the inhibition rate was 57.24%; The tumor weight of Epi group was (2.45±0.31)g, the inhibition rate was 46.27%, all groups have significant difference compared to NS group(P<0.01). The tumor weight of NS group was (4.56±0.46)g. CDI=0.9889<1. The average tumor weight of Tha+Epi group was lighter than other groups, the difference has statitistic significance .
2 The expression of bFGF mRNA and MMP-2 mRNA in tumor tissue were detected by reverse transcription polymerase chain reaction(RT-PCR) method. The expression of bFGF mRNA in Tha group, Tha+Epi group, Epi group, NS group was 0.3307±0.0463, 0.3054±0.0539, 0.6115±0.0629, 0.6732±0.1244 respectively. Comparing to NS group, there was remarkable difference in Tha group and Tha+Epi group(P<0.01), there was no difference between Epi group and NS group (P>0.05). The expression of MMP-2 mRNA in Tha group, Tha+Epi group, Epi group, NS group was 0.2835±0.0569, 0.2502±0.069, 0.6847±0.1002, 0.716±0.1126 respectively. Comparing to NS group, there was remarkable difference in Tha group and Tha+Epi group (P<0.01), there was no difference between Epi group and NS group (P>0.05).
3 The expression of bFGF protein and MMP-2 protein in tumor tissue were detected by immunohistochemical (IHC) method. The expression of bFGF protein in Tha group, Tha+Epi group, Epi group, NS group was 2.4±0.9661, 2.3±0.675, 4.5±1.1785, 5.2±0.7888 respectively. Comparing to NS group, there was significant difference in Tha group and Tha+Epi group(P<0.01), there was no difference between Epi group and NS group(P>0.05). The expression of MMP-2 protein in Tha group, Tha+Epi group, Epi group, NS group was 2.0±0.6667, 1.9±0.7379, 3.2±1.0328, 3.8±1.3166 respectively. Comparing to NS group, there was significant difference in Tha group and Tha+Epi group(P<0.01), there was no difference between Epi group and NS group(P>0.05).
Conclusions:
1 Thalidomide can inhibit the proliferation of H22 murine hepatocellular carcinoma.
2 Thalidomide can strengthen Epirubicine on H22 murine hepatocellular carcinoma proliferation inhibition, they show synergistic action.
3 Thalidomide can inhibit the proliferation in H22 murine hepatocellular carcinoma by down regulating the expression of bFGF and MMP-2.
4 This research proved Thalidomide has the effect of anti-tumor on H22 murine hepatocellular carcinoma, and inriched the treatment of H22 murine hepatocellular carcinoma through combining with Epirubicine, provided new choice in the comprehensive therapy.
方法:取40只健康雄性昆明种小鼠,无菌条件下将小鼠H22肝癌腹水瘤细胞接种到小鼠右腋皮下,0.2ml/只(2.0×106个细胞),构建荷瘤肝癌小鼠模型。接种第二天将荷瘤小鼠随机分为4组,每组10只:沙利度胺组(灌胃沙利度胺200mg/kg/日,连续给药10天)、沙利度胺联合表阿霉素组(灌胃沙利度胺200mg/kg/日,连续给药10天;腹腔注射表阿霉素50mg/m2,接种第二天给药一次)、表阿霉素组(腹腔注射表阿霉素50mg/m2,接种第二天给药一次)、生理盐水对照组(灌胃生理盐水0.3ml/日,连续给药10天),停药次日脱臼处死,完整剥离瘤组织,称重,计算抑瘤率及两药相互作用系数。分别提取各组瘤细胞总RNA,鉴定其完整性及含量,采用半定量RT-PCR方法检测肿瘤组织内bFGF mRNA、MMP-2 mRNA的表达水平,应用免疫组化方法检测肿瘤组织内bFGF、MMP-2蛋白的表达水平。用SPSS15.0统计学软件进行数据处理,各组数据以均值±标准差的形式表示。
结果:
1沙利度胺组瘤重为(3.67±0.44)g,抑瘤率为19.52%,沙利度胺联合表阿霉素组瘤重为(1.95±0.24)g,抑瘤率为57.24%,表阿霉素组瘤重为(2.45±0.31)g,抑瘤率为46.27%,均与生理盐水对照组相比有统计学意义(P<0.01)。生理盐水对照组瘤重为(4.56±0.46)g。两药相互作用系数(CDI)=0.9889<1,沙利度胺联合表阿霉素组平均瘤重最低,与沙利度胺组、表阿霉素组相比均有统计学意义(P<0.01)。
2采用半定量RT-PCR的方法检测各组肿瘤组织内bFGF mRNA、MMP-2 mRNA的表达水平。沙利度胺组bFGF mRNA表达相对值为0.3307±0.0463,沙利度胺联合表阿霉素组bFGF mRNA表达相对值为0.3054±0.0539,均与生理盐水对照组相比有统计学差异(P<0.01)。生理盐水对照组bFGF mRNA表达相对值为0.6732±0.1244,表阿霉素组bFGF mRNA表达相对值为0.6113±0.0629。生理盐水对照组和表阿霉素组相比无统计学意义(P>0.05)。沙利度胺组MMP-2 mRNA表达相对值为0.2835±0.0569,沙利度胺联合表阿霉素组MMP-2 mRNA表达相对值为0.2502±0.069,均与生理盐水对照组相比有统计学意义(P<0.01)。生理盐水对照组MMP-2 mRNA表达相对值为0.716±0.1126,表阿霉素组MMP-2 mRNA表达相对值为0.6847±0.1002,生理盐水对照组和表阿霉素组相比无统计学意义(P>0.05)。
3应用免疫组化方法检测各组肿瘤组织内bFGF、MMP-2蛋白的表达水平。沙利度胺组bFGF表达值为2.4±0.9661,沙利度胺联合表阿霉素组bFGF表达值为2.3±0.675,均与生理盐水对照组相比有统计学意义(P<0.01)。生理盐水对照组bFGF表达值为5.2±0.7888,表阿霉素组bFGF表达值为4.5±1.1785,表阿霉素组和生理盐水对照组相比无统计学意义(P>0.05)。沙利度胺组MMP-2表达值为2.0±0.6667,沙利度胺联合表阿霉素组MMP-2表达值为1.9±0.7379,均与生理盐水对照组相比有统计学意义(P<0.01)。生理盐水对照组MMP-2表达值为3.8±1.3166,表阿霉素组MMP-2表达值为3.2±1.0328,表阿霉素组和生理盐水对照组相比无统计学意义(P>0.05)。
结论:
1沙利度胺具有抑制小鼠H22肝癌移植瘤增殖的作用。
2沙利度胺可协同表阿霉素对小鼠H22肝癌移植瘤增殖的抑制作用。
3沙利度胺可能通过下调bFGF、MMP-2的表达而发挥抑制小鼠H22肝癌移植瘤增殖作用。
4本实验证实了沙利度胺具有抗肝癌作用,并通过沙利度胺联合抗肿瘤药物表阿霉素的实验研究丰富了肝癌的治疗,为肝癌的综合治疗提供了新的选择。
Objective: Established an H22 mice hepatocellular carcinoma model, the effict of Thalidomide(Tha) combined with Epirubicine(Epi) on H22 murine hepatocellular carcinoma was observed, the aim was to explore the possible mechanism of thalidomide anti-tumor.
Methods: Fourty healthy male kunming mice were transplanted by right armpit injection of H22 tumor cells in asepsis conditions, 0.2ml per mice(2.0×106 cells). Then these mice were randomly divided into four groups, ten mice in each group. Tha group (oral Tha 200mg/kg/d, d1-10), Tha+Epi group (oral Tha 200mg/kg/d, d1-10; Epirubicine intraperitoneal injection, 50mg/m2, d1), Epi group (Epirubicine intraperitoneal injection,50mg/m2, d1), Normal Saline group (oral normal saline 0.3ml/d, d1-10). Ten days later all mice were killed and the tumor were taken and weighed. Then the ratio of tumor suppression could be calculated, coefficient of drug interaction(CDI) was calculated. The expression of bFGF mRNA and MMP-2 mRNA were examined by reverse transcription polym-erase chain reaction(RT-PCR). The expression of bFGF, MMP-2 in tumor tissue was detected by using immunohistochemical (IHC) method. All data was showed by Mean±SD. The data was analyzed with the SPSS15.0 software.
Results:
1 The tumor weight of Tha group was (3.67±0.44)g, the inhibition rate was 19.52%; The tumor weight of Tha+Epi group was (1.95±0.24)g, the inhibition rate was 57.24%; The tumor weight of Epi group was (2.45±0.31)g, the inhibition rate was 46.27%, all groups have significant difference compared to NS group(P<0.01). The tumor weight of NS group was (4.56±0.46)g. CDI=0.9889<1. The average tumor weight of Tha+Epi group was lighter than other groups, the difference has statitistic significance .
2 The expression of bFGF mRNA and MMP-2 mRNA in tumor tissue were detected by reverse transcription polymerase chain reaction(RT-PCR) method. The expression of bFGF mRNA in Tha group, Tha+Epi group, Epi group, NS group was 0.3307±0.0463, 0.3054±0.0539, 0.6115±0.0629, 0.6732±0.1244 respectively. Comparing to NS group, there was remarkable difference in Tha group and Tha+Epi group(P<0.01), there was no difference between Epi group and NS group (P>0.05). The expression of MMP-2 mRNA in Tha group, Tha+Epi group, Epi group, NS group was 0.2835±0.0569, 0.2502±0.069, 0.6847±0.1002, 0.716±0.1126 respectively. Comparing to NS group, there was remarkable difference in Tha group and Tha+Epi group (P<0.01), there was no difference between Epi group and NS group (P>0.05).
3 The expression of bFGF protein and MMP-2 protein in tumor tissue were detected by immunohistochemical (IHC) method. The expression of bFGF protein in Tha group, Tha+Epi group, Epi group, NS group was 2.4±0.9661, 2.3±0.675, 4.5±1.1785, 5.2±0.7888 respectively. Comparing to NS group, there was significant difference in Tha group and Tha+Epi group(P<0.01), there was no difference between Epi group and NS group(P>0.05). The expression of MMP-2 protein in Tha group, Tha+Epi group, Epi group, NS group was 2.0±0.6667, 1.9±0.7379, 3.2±1.0328, 3.8±1.3166 respectively. Comparing to NS group, there was significant difference in Tha group and Tha+Epi group(P<0.01), there was no difference between Epi group and NS group(P>0.05).
Conclusions:
1 Thalidomide can inhibit the proliferation of H22 murine hepatocellular carcinoma.
2 Thalidomide can strengthen Epirubicine on H22 murine hepatocellular carcinoma proliferation inhibition, they show synergistic action.
3 Thalidomide can inhibit the proliferation in H22 murine hepatocellular carcinoma by down regulating the expression of bFGF and MMP-2.
4 This research proved Thalidomide has the effect of anti-tumor on H22 murine hepatocellular carcinoma, and inriched the treatment of H22 murine hepatocellular carcinoma through combining with Epirubicine, provided new choice in the comprehensive therapy.
引文
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29 Kim M, Sata M, Ueno, et al. Basic fibroblast growth factor regulates proliferation and motility of human hepatoma cells by a autocrine mechanism. J Hepatol, 1997, 27:677~687
30 Sasada R, Kurokawa J, Iwane M, et al. Transformation of mouse BACB/C 3T3 cells with hu-man basic fibroblast growth factor cDNA. Mol Cell Biol , 1998, (8) :588~594
31 韩斌,王广田,庞志刚,等. bFGF在肝细胞癌中的表达及意义.胃肠病学和肝病学杂志, 2002, 11(2):163~165
32 Imura S, Miyake H, Izumi K, et al. Correlation of vascularendothelial cell proliferation with microvessel density and exp ression of vascular endothelial growth factor and basic fibroblast growth factor in hepatocellular carcinoma. J Med Invest, 2004, 51 (34) : 202
33 安家泽,窦科峰,李开宗,等. bFGF基因表达与肝细胞癌侵袭性的关系. 第四军医大学学报, 1999, 20 (4):342~345
34 Goddard JC, Sutton CD, Furness PN, et al. Microvessel density at presentation predicts subsequent muscle invasion in superficial bladder cancer. Clin Cancer Res, 2003, 9(7): 583~586
35 孙惠川,汤钊猷,王鲁,等. 肝细胞癌患者血浆VEGF、sVCAM和bFGF浓度的研究与术后短期复发的相关性. 癌症, 2001, 21(6):455~457
36 李西平,刘叙仪,王洁,等. 反应停对人肺腺癌亲代及耐顺铂细胞系碱性成纤维细胞生长因子表达影响. 中国呼吸与危重监护杂志, 2003, 2 (1):40~43
37 Turpeenniemi-Hujanen T. Gelatinases (MMP-2 and-9) and their natural inhibitors as prognostic indicators in solid cancers. Biochimie, 2005, 87:287~297
38 Collen A, Hanemaaijer R, Lupu F, et al. Membrane-type matrix metallop roteinase-mediated angiogenesis in a fibrin collagen matrix. Blood, 2003, 101:1810~1817
39 Ohno-Matsui K, Uetama T, Yoshida T, et al. Reduced retinal angiogenesis in MMP-2-deficient Mice. Invest Ophthalmol Vis Sci, 2003, 44:5370~5375
40 郭荣平,钟崇,石明,等. 血管内皮生长因子和基质金属蛋 白酶-2在肝细胞癌中的表达及临床意义.中华肿瘤杂志, 2006, 28 (4):285~288
1 Okada-Ban M, Thiery JP, Jouanneau J, et al. Fibroblast growth factor2. Int Biohem, 2000, 32 (3):263~267
2 Szebenyi G, Fallon JF. Fibroblast growth factors as multifunctional signaling facors. Int Rev Cytol, 1999, 185:45~106
3 Song H, Kwon K, Lim S, et al. Trans fection of Mesenchymal Stem Cells with the FGF-2 Gene ImprovesTheir Survival under Hypoxic Conditions. Molecules and Cells, 2005, 19(3): 402~407
4 Kumar R, Yoneda J, Bucana CD, et al. Regulation of distinct steps of angiogenesis by different angiogenic molecules. J Int J Oncol, 1998, 12 (4):749~757
5 Kim M, Sata M, Ueno, et al. Basic fibroblast growth factor regulates proliferation and motility of human hepatoma cells by a autocrine mechanism. J Hepatol, 1997, 27:677~687
6 Sasada R, Kurokawa J, Iwane M, et al. Transformation of mouse BACB/C 3T3 cells with hu-man basic fibroblast growth factor cDNA. Mol Cell Biol, 1998, 8:588~594
7 El-Assal ON, Yamanoi A, Ono T, et al. The clinicopathol- ogical significance of heparanase and basic fibroblast growth factor expressions in hepatocellular carcinoma. Clin Cancer Res, 2001, 7 (5):1299~1305
8 韩斌,王广田,庞志刚等. bFGF在肝细胞癌中的表达及意义.胃肠病学和肝病学杂志, 2002, 11(2) :163~165
9 张华,郭崇洁,景朋,等. 无血清培养人肝癌细胞增生率的变化和bFGF在细胞中的表达. 首都医科大学学报,2003, 24(1):1~3
10 Ogasawara S, Yano H, Iemura A, et al. Expression of basic fibroblast growthfactor and its receptors and their relationship to proliferation of human hepatocellular carcinoma cell lines. Hepatology, 1996, 24:198~205
11 Folkman J, Shing Y. Angiogenesis J. J Biol Chem, 1992, 267 (16):10931~10934
12 韩斌,梁冰,王广田,等. 肝细胞癌组织中碱性成纤维细胞生长因子和微血管密度检测. 郑州大学学报(医学版), 2006, 7(4):704~706
13 Imura S, Miyake H, Izumi K, et al. Correlation of vascular endothelial cell proliferation with microvessel density and exp ression of vascular endothelial growth factor and basic fibroblast growth factor in hepatocellular carcinoma. J Med Invest, 2004, 51(34):202
14 Yoshiji H, Kuriyama S, Yoshii J, et al. Synergistic effect of basic fibroblast growth factor and vascular endot helial growt hfactor in murine hepatocellular carcinoma. J Hepatology, 2002, 35:834~842
15 Pepper MS, Mandriota SJ, JeltschM, et al. Vascular endo- thelial growth factor(VEGF)-C synergizes with basic fibroblast growth factor and VEGF in the induction of angiogenesis in vitro and alters endothelial cell extracellular propeolytic activity. J Cell Physiol , 1998, 177:439~452
16 Seghezzi G, Patel S, Ren CJ, et al. Fibroblast growth factor2 (FGF2) induces vascular expression in the endothelial cell of forming capillaries:an autocrine mechanism contributing to angiogenesis. J Cell Biol, 1998, 141:1659~1673
17 Saadeh PB, Mehrara BJ, Steinbrech DS, et al. Mechanisms of fibroblast growth factor2 modulation of vascular endithelial growth factor expression by osteoblastic cell. Endocrinology, 2000, 14:2075~2083
18 Moon WS, Rhyu KH, Kang MJ, et al. Overexpression of VEGF and angiopoietin2:a key to high vascularity of hepatocellular carcinoma. J Mod Pathol, 2003, 36(6): 552~557
19 Yamaguchi R. Yano H, lmura A, et, al. Epression of vescular endothelial growth factor in human hepotacellular carcinoma. Hepatology, 1998, 28:68~77
20 安家泽,窦科峰,李开宗,等. bFGF基因表达与肝细胞癌侵袭性的关系. 第四军医大学学报, 1999, 20(4):342~345
21 于德新,高志芹,王滨,等. 肝细胞癌VEGF、bFGF与凋亡相关蛋白表达的关系. 肿瘤防治研究, 2005, 32(5):276~278
22 Goddard JC, Sutton CD, Furness PN, et al. Microvessel density at presentation predicts subsequent muscle invasion in superficial bladder cancer. Clin Cancer Res, 2003, 9(7): 583-586
23 孙惠川,汤钊猷,王鲁,等. 肝细胞癌患者血浆VEGF、VCAM和bFGF浓度的研究与术后短期复发的相关性. 癌症, 2001, 21(6):455-457
24 Folkman J, Daniel F, Jerome P, et al. Elevated levels of an angiogenic peptide, basic fibroblast growth factor, in the urine of patients with a wide spectrum of cancers. J Natl Cancer Inst, 1994, 86(5):356~361
25 David M, Nanus, Bernd J, et al. Expression of basic fibroblast growth factor in primary human renal tumors:correlation with poor survial. J Natl Cancer Inst, 1993, 85(19): 1597~1599
26 周廷冲.多肽生长因子基础与临床.北京,中国科学技术出 版社, 1992:45~47
27 Lin YC, Shun CT, Wu MS, et al. A novel anticancer effect of thalidomide:inhibition of intercellular adhesion molecule- 1-mediated cell invasion and metastasis through suppression of nuclear factor-kappaB. J Clin Cancer Res, 2006, 12 (23):7165~7273
2 Kebrel RS. Clinical trials of antiangiogenic drugs: opportunities porblems and assessment of initial results. J clin oncol, 2001, (19):45~51
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8 Groxtall JD, Flower RJ. Lipocortin1 mediates dexamet hasone induced growth arres of the A549 lung adeno carcinoma cell line. J Proc Natl Acad Sci USA, 1992, 89(8): 3571~3575
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11 Bisacchi D, Benelli R, Vanzetto C, et al. Anti-angiogensis and angioprevention mechanisms, problems and perspectives perspectives. J Cancer Detection and Prevention, 2003, 27(3):229~266
12 El-Assal ON, Yamanoi A, Ono T, et al. The clinicopatholo- gical significance of heparanase and basic fibroblast growth factor expressions in hepatocellular carcinoma. Clin Cancer Res, 2001, 7(5):1299~1305
13 Musto P. Thalidomide therapy for myelodysplastic syndromes:current status and future perspectives. J Leukemia Research, 2004, 28(4):325~356
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18 黄承钰,曾令福,徐亚林,等. 小鼠实体型肝癌H22移植实验 条 件 探 讨 . 临 床 与 实 验 病 理 学 杂 志 ,1997, 13(2):188~189
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27 Slodkowska J, Sikora J, Roszkowski-Sliz K, et al. Expression of vascular endothelial growth factor and basic fibroblast growth factor receptors in lung cancer. J Analyt Quant Cytol Histol,2000,22(5):398~402
28 Poon RT, Fan ST, Wong J. Clinical implications of circulating angiogenic factors in cancer patients. J Clin Oncol, 2001, 19(4):1207~1225
29 Kim M, Sata M, Ueno, et al. Basic fibroblast growth factor regulates proliferation and motility of human hepatoma cells by a autocrine mechanism. J Hepatol, 1997, 27:677~687
30 Sasada R, Kurokawa J, Iwane M, et al. Transformation of mouse BACB/C 3T3 cells with hu-man basic fibroblast growth factor cDNA. Mol Cell Biol , 1998, (8) :588~594
31 韩斌,王广田,庞志刚,等. bFGF在肝细胞癌中的表达及意义.胃肠病学和肝病学杂志, 2002, 11(2):163~165
32 Imura S, Miyake H, Izumi K, et al. Correlation of vascularendothelial cell proliferation with microvessel density and exp ression of vascular endothelial growth factor and basic fibroblast growth factor in hepatocellular carcinoma. J Med Invest, 2004, 51 (34) : 202
33 安家泽,窦科峰,李开宗,等. bFGF基因表达与肝细胞癌侵袭性的关系. 第四军医大学学报, 1999, 20 (4):342~345
34 Goddard JC, Sutton CD, Furness PN, et al. Microvessel density at presentation predicts subsequent muscle invasion in superficial bladder cancer. Clin Cancer Res, 2003, 9(7): 583~586
35 孙惠川,汤钊猷,王鲁,等. 肝细胞癌患者血浆VEGF、sVCAM和bFGF浓度的研究与术后短期复发的相关性. 癌症, 2001, 21(6):455~457
36 李西平,刘叙仪,王洁,等. 反应停对人肺腺癌亲代及耐顺铂细胞系碱性成纤维细胞生长因子表达影响. 中国呼吸与危重监护杂志, 2003, 2 (1):40~43
37 Turpeenniemi-Hujanen T. Gelatinases (MMP-2 and-9) and their natural inhibitors as prognostic indicators in solid cancers. Biochimie, 2005, 87:287~297
38 Collen A, Hanemaaijer R, Lupu F, et al. Membrane-type matrix metallop roteinase-mediated angiogenesis in a fibrin collagen matrix. Blood, 2003, 101:1810~1817
39 Ohno-Matsui K, Uetama T, Yoshida T, et al. Reduced retinal angiogenesis in MMP-2-deficient Mice. Invest Ophthalmol Vis Sci, 2003, 44:5370~5375
40 郭荣平,钟崇,石明,等. 血管内皮生长因子和基质金属蛋 白酶-2在肝细胞癌中的表达及临床意义.中华肿瘤杂志, 2006, 28 (4):285~288
1 Okada-Ban M, Thiery JP, Jouanneau J, et al. Fibroblast growth factor2. Int Biohem, 2000, 32 (3):263~267
2 Szebenyi G, Fallon JF. Fibroblast growth factors as multifunctional signaling facors. Int Rev Cytol, 1999, 185:45~106
3 Song H, Kwon K, Lim S, et al. Trans fection of Mesenchymal Stem Cells with the FGF-2 Gene ImprovesTheir Survival under Hypoxic Conditions. Molecules and Cells, 2005, 19(3): 402~407
4 Kumar R, Yoneda J, Bucana CD, et al. Regulation of distinct steps of angiogenesis by different angiogenic molecules. J Int J Oncol, 1998, 12 (4):749~757
5 Kim M, Sata M, Ueno, et al. Basic fibroblast growth factor regulates proliferation and motility of human hepatoma cells by a autocrine mechanism. J Hepatol, 1997, 27:677~687
6 Sasada R, Kurokawa J, Iwane M, et al. Transformation of mouse BACB/C 3T3 cells with hu-man basic fibroblast growth factor cDNA. Mol Cell Biol, 1998, 8:588~594
7 El-Assal ON, Yamanoi A, Ono T, et al. The clinicopathol- ogical significance of heparanase and basic fibroblast growth factor expressions in hepatocellular carcinoma. Clin Cancer Res, 2001, 7 (5):1299~1305
8 韩斌,王广田,庞志刚等. bFGF在肝细胞癌中的表达及意义.胃肠病学和肝病学杂志, 2002, 11(2) :163~165
9 张华,郭崇洁,景朋,等. 无血清培养人肝癌细胞增生率的变化和bFGF在细胞中的表达. 首都医科大学学报,2003, 24(1):1~3
10 Ogasawara S, Yano H, Iemura A, et al. Expression of basic fibroblast growthfactor and its receptors and their relationship to proliferation of human hepatocellular carcinoma cell lines. Hepatology, 1996, 24:198~205
11 Folkman J, Shing Y. Angiogenesis J. J Biol Chem, 1992, 267 (16):10931~10934
12 韩斌,梁冰,王广田,等. 肝细胞癌组织中碱性成纤维细胞生长因子和微血管密度检测. 郑州大学学报(医学版), 2006, 7(4):704~706
13 Imura S, Miyake H, Izumi K, et al. Correlation of vascular endothelial cell proliferation with microvessel density and exp ression of vascular endothelial growth factor and basic fibroblast growth factor in hepatocellular carcinoma. J Med Invest, 2004, 51(34):202
14 Yoshiji H, Kuriyama S, Yoshii J, et al. Synergistic effect of basic fibroblast growth factor and vascular endot helial growt hfactor in murine hepatocellular carcinoma. J Hepatology, 2002, 35:834~842
15 Pepper MS, Mandriota SJ, JeltschM, et al. Vascular endo- thelial growth factor(VEGF)-C synergizes with basic fibroblast growth factor and VEGF in the induction of angiogenesis in vitro and alters endothelial cell extracellular propeolytic activity. J Cell Physiol , 1998, 177:439~452
16 Seghezzi G, Patel S, Ren CJ, et al. Fibroblast growth factor2 (FGF2) induces vascular expression in the endothelial cell of forming capillaries:an autocrine mechanism contributing to angiogenesis. J Cell Biol, 1998, 141:1659~1673
17 Saadeh PB, Mehrara BJ, Steinbrech DS, et al. Mechanisms of fibroblast growth factor2 modulation of vascular endithelial growth factor expression by osteoblastic cell. Endocrinology, 2000, 14:2075~2083
18 Moon WS, Rhyu KH, Kang MJ, et al. Overexpression of VEGF and angiopoietin2:a key to high vascularity of hepatocellular carcinoma. J Mod Pathol, 2003, 36(6): 552~557
19 Yamaguchi R. Yano H, lmura A, et, al. Epression of vescular endothelial growth factor in human hepotacellular carcinoma. Hepatology, 1998, 28:68~77
20 安家泽,窦科峰,李开宗,等. bFGF基因表达与肝细胞癌侵袭性的关系. 第四军医大学学报, 1999, 20(4):342~345
21 于德新,高志芹,王滨,等. 肝细胞癌VEGF、bFGF与凋亡相关蛋白表达的关系. 肿瘤防治研究, 2005, 32(5):276~278
22 Goddard JC, Sutton CD, Furness PN, et al. Microvessel density at presentation predicts subsequent muscle invasion in superficial bladder cancer. Clin Cancer Res, 2003, 9(7): 583-586
23 孙惠川,汤钊猷,王鲁,等. 肝细胞癌患者血浆VEGF、VCAM和bFGF浓度的研究与术后短期复发的相关性. 癌症, 2001, 21(6):455-457
24 Folkman J, Daniel F, Jerome P, et al. Elevated levels of an angiogenic peptide, basic fibroblast growth factor, in the urine of patients with a wide spectrum of cancers. J Natl Cancer Inst, 1994, 86(5):356~361
25 David M, Nanus, Bernd J, et al. Expression of basic fibroblast growth factor in primary human renal tumors:correlation with poor survial. J Natl Cancer Inst, 1993, 85(19): 1597~1599
26 周廷冲.多肽生长因子基础与临床.北京,中国科学技术出 版社, 1992:45~47
27 Lin YC, Shun CT, Wu MS, et al. A novel anticancer effect of thalidomide:inhibition of intercellular adhesion molecule- 1-mediated cell invasion and metastasis through suppression of nuclear factor-kappaB. J Clin Cancer Res, 2006, 12 (23):7165~7273