Survivin反义寡核苷酸对肝癌细胞株SMMC-7721增殖的影响

目的:应用反义寡核苷酸技术研究凋亡抑制蛋白Survivin反义寡核苷酸对肿瘤细胞增殖的影响。方法:根据Survivin序列合成其反义寡核苷酸序列(Survivin-ASODN)。培养肝癌细胞株SMMC-7721,用荧光素标记的Survivin-ASODN转染,并设立空白对照、空脂质体对照及正义寡核苷酸(SODN)对照组;用PCR方法检测Survivin基因表达水平;通过MTT方法分析Survivin-ASODN转染对肝癌细胞增殖的影响。结果:荧光素标记的Survivin-ASODN在SMMC-7721细胞中的转染率达到60%以上。SMMC-7721细胞分别用100、200、300、400和600nmol·L1ASODN转染后,Survivin基因表达水平下降,细胞增殖抑制率24h时分别为(3.87±1.67)%、(9.21±2.21)%、(15.21±3.18)%、(21.32±3.43)%和(32.62±3.74)%,而300nmol·L1ASODN组48h时细胞生长抑制率为(29.21±4.12)%,72h时抑制率为(44.21±3.32)%;ASODN转染组细胞增殖抑制率与空白对照组、空脂质体组及SODN组比较,差异均有统计学意义(P<0.05)。结论:Survivin-ASODN对细胞增殖有抑制作用,且呈现剂量依赖性;并且随着作用时间的延长,其抑制效果增强。

Effect of Survivin-ASODN on growth of hepatocellular carcinoma cells

Objective To study the influence of anti-sense oligonucleotides(ASODN) of Survivin on carcinoma cell proliferation by the technique of ASODN.Methods Survivin-ASODN was synthesized according to Survivin gene sequence.The SMMC-7721 cells were transfected with fluorescence labled Survivin-ASODN.At the same time,blank control,lipofectamine control and SODN control groups were set up.The Survivin gene expression levels were measured by PCR.The proliferation inhibitory rates of Survivin-ASODN transfected cells were analyzed with MTT method.Results The transfection rate reached 60% of fluorescence labled Survivin-ASODN in SMMC-7721 cells.After transfected with ASODN at 100,200,300,400 and 600 nmol·L-1,the Survivin gene expression levels were decreased.The inhibitory rates were(3.87±1.67)%,(9.21±2.21)%,(15.21±3.18)%,(21.32±3.43)%,and(32.62±3.74)%,respectively,at 24 h after transfection.In 300 nmol·L-1 ASODN group,the inhibitory rates were(29.21±4.12)% at 48 h and(44.21±3.32)% at 72 h.Compared with blank control group,lipofectamine group and SODN group,the inhibitory rates in ASODN group showed significant difference(P<0.05).Conclusion Survivin-ASODN has inhibitory effect on cell proliferation in dose-dependent and time-dependent manner.