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血管生成素-1在体外对人胃癌细胞生物学作用的研究
作者:杨柳 欧希龙 颜芳 关云艳 杭程 陈慧娟 陈国胜 
单位:东南大学,临床医学院,江苏,南京,210009
关键词:血管生成素-1 血管内皮生长因子165 Ki-67 胃癌 增殖 凋亡 
分类号:R735.2, R-33
出版年·卷·期(页码):2008·27·第二期(116-121)
摘要:

目的:探讨人血管生成素-1(Ang1)及其与血管内皮生长因子165(VEGF165)共同作用对人胃癌细胞的生物学作用,研究其在肿瘤发生中的作用机制.方法:应用复制缺陷型腺病毒(Ad)-绿色荧光蛋白(GFP)、Ad-Ang1、Ad-VEGF165和Ad-Ang1+Ad-VEGF165/2转染人胃癌细胞株MGC-803,使用MTT比色法分析它们对胃癌细胞增殖的影响;通过流式细胞仪分析血浆饥饿时其对凋亡的影响;使用免疫细胞化学法检测Ki-67的表达;应用RT-PCR方法半定量检测bcl-2 mRNA、bax mRNA的表达情况.结果:MTT结果显示24、48和72 h Ad-Ang1转染组、Ad-VEGF165转染组和Ad-Ang1+Ad-VEGF165/2转染组吸光度均明显高于Ad-GFP组及对照组(P<0.05);Ad-GFP组与对照组相比无显著差异(P>0.05);Ad-Ang1+Ad-VEGF165/2转染组吸光度明显高于Ad-Ang1转染组和Ad-VEGF165转染组(P<0.05).流式细胞仪检测结果显示Ad-Ang1、Ad-VEGF165及Ad-Ang1+Ad-VEGF165/2处理组与对照组及空转染组相比均能够显著抑制细胞凋亡(P<0.01).免疫细胞化学法显示各转染组Ki-67表达强度均显著高于对照组和Ad-GFP组(P<0.01).RT-PCR结果显示bcl-2 mRNA在各转染组中的表达要明显高于对照组(P<0.05);bax mRNA在各转染组中的表达要明显低于对照组(P<0.05).结论:Ang1、VEGF165和Ang1+VEGF165/2能够明显促进人胃癌细胞MGC-803增殖,抑制血浆饥饿时的凋亡,Ang1+VEGF165/2组的作用要显著强于Ang1组和VEGF165组.

Objective To explore the biological effect of angiopoietin-1(Ang1)and the interaction of Ang1 and VEGF165 on human gastric cancer cell line MGC-803,and study their mechanism in tumorigenesis.Methods The cell line MGC-803 was transfected with Ad-GFP,Ad-Ang1,Ad-VEGF165 and Ad-Ang1+Ad-VEGF165/2.The proliferation and apoptosis of MGC-803 was measured by MTT colorimetry and flow cytometer respectively.Expression of Ki-67 was measured by immunocytochemistry.The expression of bcl-2 mRNA,bax mRNA was detected by reverse transcription-polymerase chain reaction(RT-PCR).Results Ang1,VEGF165 and Ang1+VEGF165/2 transfected MGC-803 successfully.Proliferations of MGC803 cells cultured in Ad-Ang1 group,Ad-VEGF165 group and Ad-Ang1+Ad-VEGF165/2 group were higher than Ad-GFP group and controlled group(P&lt;0.05).Proliferation of MGC-803 cells cultured in Ad-Ang1+ Ad-VEGF165/2 group was higher than Ad-Ang1 group and Ad-VEGF165 group(P&lt;0.05).Ang1,VEGF165 and Ang1+VEGF-165/2 could significantly inhibit cell apoptosis comparing with Ad-GFP group and controlled group(P&lt;0.01).Comparing with Ad-GFP group and controlled group,Ki-67 expressing intensity in every tranfected group greatly increased(P&lt;0.01).Expression of bcl-2 mRNA increased in every tranfected group(P&lt;0.01) and expression in Ad-Ang1+ Ad-VEGF165/2 group was strongest.Expression of bax mRNA was reverse.Conclusions Ang1,VEGF165 and Ang1+VEGF165/2 can significantly promote proliferation and inhibit apoptosis of human gastric cancer cell line MGC 803 in vitro.The effect of Ang1+VEGF165/2 on gastric cancer cell line MGC-803 is stronger than Ang1 and VEGF165.

参考文献:

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