摘要目的 探讨survivin反义脱氧寡核苷酸(ASODN)对胃癌细胞株BGC-823凋亡、增殖的作用及分子机制.方法 分别以survivin ASODN-1、survivin ASODN-2和survivin ASODN-3转染人胃癌BGC-823细胞株,采用共聚焦显微镜检测转染率,四甲基偶氮唑蓝(MTF)法检测细胞生长活性,流式细胞术检测细胞凋亡指数、增殖指数和细胞周期分布及survivin、血管内皮生长因子(VEGF)、Smac/DIABLO蛋白表达改变,逆转录聚合酶链反应检测survivin、VEGF、Smac/DIABLO mRNA表达改变.结果 转染各序列survivin ASODN均可下调survivin蛋白的表达,且以ASODN-2作用最为显著.以600 nmol/L survivin ASODN-2转染BGC-823细胞48 h后,G0/G1期细胞比例[(72.25±2.95)%]明显高于空脂质体对照组[(56.25±0.75)%,均P＜0.05],凋亡指数[(11.31±0.38)%]明显高于空脂质体对照组[(1.62±0.36)%,均P＜0.05],增殖指数[(27.77±2.97)%]低于空脂质体对照组[(43.80±0.80)%,均P＜0.05].转染后survivin mRNA及蛋白表达(0.523±0.091,0.733±0.009)低于空脂质体对照组(0.861±0.047,0.997±0.233;均P＜0.05),VEGF mRNA及蛋白表达(0.519±0.076,0.75±0.006)低于空脂质体对照组(0.779±0.059,1.000±0.01;均P＜0.05),Smac/DIABLOmRNA及蛋白表达(0.899±0.113,1.637±0.023)高于空脂质体对照组(0.558±0.041,1.000±0.049;均P＜0.05).结论 survivin ASODN具有诱导胃癌细胞凋亡、抑制细胞增殖的作用,其作用是通过下调survivin和VEGF、上调Smac/DIABLO基因表达实现的.

abstractsObjective To explore the effects of antisense oligodeoxynucleotides (ASODN)on proliferation and apoptosis in gastric cancer cell line BGC-823 cells and the molecular mechanisms induced by ASODN.Methods survivin ASODN-1, survivin ASODN-2 and survivin ASODN-3 were transfected into BGC-823 cells by LipofectamineTM 2000 transfection reagent.The growth activity of BGC-823 cells was detected by MTT assay.Apoptosis index (AI), proliferation index ( PI), cell cycle and expressions of survivin, VEGF and Smac/DIABLO proteins were detected by flow cytometry (FCM).The changes of survivin mRNA, VEGF mRNA and Smac/DIABLO mRNA were detected by RT-PCR.Results The expression of survivin was down-regulated by the three ASODN sequences, especially the ASODN-2 was best.At 48 hours after transfection with 600 nmol/L survivin ASODN-2, the cells in G1/G0 phase were significantly increased [(72.25 ± 2.95 ) %], apoptotic index increased [( 11.31 ± 0.38 ) %], proliferation index decreased [(27.77 ± 2.97 ) %], compared with those in the control group [( 56.25 ± 0.75 ) %,(1.62 ±0.36)%, (43.80 ±0.80)%, all P ＜ 0.05].The survivin mRNA and protein levels (0.523 ±0.091,0.733 ±0.009) were down-regulated compared with those in the control group (0.861 ±0.047,0.997 ± 0.233 ), VEGF (0.519 ± 0.076, 0.75 ± 0.006) were down-regulated compared with those in the control group (0.779 ± 0.059, 1.000 ± 0.01 ), while those of Smac/DIABLO (0.899 ± 0.113, 1.637 ±0.023) were up-regulated compared with those in the control group (0.558 ± 0.041, 1.000 ± 0.049, all P＜0.05).Conclusions Survivin ASODN can induce apoptosis and inhibit the proliferation of gastric cancer cell line BGC-823 cells.Those effects are induced through up-regulation of Smac/DIABLO and downregulation of survivin and VEGF expression simultaneously.