survivin反义脱氧寡核苷酸对胃癌细胞增殖与凋亡的影响  

Effect of antisurvivin oligodeoxynucleotides on proliferation and apoptosis in gastric cancer cell line BGC-823 and the molecular mechanism

作　　者：李勇[1] 范立侨[1] 孙艳[2] 赵群[1] 檀碧波[1] 刘羽[1] 

机构地区：[1]河北医科大学第四医院外三科,石家庄 050011 [2]河北医科大学第三医院妇产科

出　　处：《中华肿瘤杂志》2011年第4期265-269,共5页Chinese Journal of Oncology

基　　金：河北省普通高校强势特色学科资助项目（冀教高[2005]52）;河北省科技厅科研基金（08276101D-38）

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survivinASODN-1、survivinASODN-2和survivinASODN-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（MTT）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／DIABLOmRNA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600nmoULsurvivinASODN-2转染BGC-823细胞48h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（0．86l±0．047，0．997±0．233；均P〈0．05），VEGFmRNA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。Objective 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 MTI assay. Apoptosis index （AI）, proliferation index （PI）, cell cycle and expressions of survivin, VEGF and Smae/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 down- regulation of survivin and VEGF expression simultaneously.

关 键 词：胃肿瘤 生存素 血管内皮生长因子 Smac/DIABLO基因 寡核苷酸类 反义 BGC-823细胞 增殖 凋亡 

分 类 号：R735.2[医药卫生—肿瘤]