血管生成素-1在体外对人胃癌细胞生物学作用的研究  

Study of biological effect of angiopoietin-1 on human gastric cancer cell in vitro

作　　者：杨柳[1] 欧希龙[1] 颜芳[1] 关云艳[1] 杭程[1] 陈慧娟[1] 陈国胜[1] 

机构地区：[1]东南大学临床医学院,江苏南京210009

出　　处：《东南大学学报（医学版）》2008年第2期116-121,共6页Journal of Southeast University(Medical Science Edition)

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 transcriptionpolymerase chain reaction （RT-PCR）. Results Ang1, VEGF165 and Ang1 ＋ VEGF165/2 transfected MGC-803 successfully. Proliferations of MGC-803 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 〈 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 〈 0.05）. Ang1 ,VEGF165 and Ang1 ＋ VEGF-165/2 could significantly inhibit cell apoptosis comparing with Ad-GFP group and controlled group （P 〈 0.01 ）. Comparing with Ad-GFP group and controlled group, Ki-67 expressing intensity in every tranfected group greatly increased （P 〈0.01 ）. Expression of bcl-2 mRNA increased in every tranfected group（ P 〈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.

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[医药卫生—肿瘤] R-33[医药卫生—临床医学]