生存素反义寡核苷酸对人子宫内膜癌细胞增殖的抑制作用

Inhibitory effects of survivin antisense oligonucleotide on human endometrial carcinoma cells

目的探讨生存素(survivin)反义寡核苷酸(antisense oligonucleotide,ASODN)对人子宫内膜癌细胞HEC-1B增殖的抑制作用。方法实验分空白对照组(control组)、单纯脂质体对照组(Lip组)、正义链转染对照组(SODN组)、ASODN转染组(ASODN组)4组。人工合成正、反义寡核苷酸,经脂质体将survivin正、反义寡核苷酸转染入子宫内膜癌细胞48 h后收集各组细胞。免疫印迹(Western blot)法检测各组细胞生存素表达情况,流式细胞仪检测各组细胞凋亡率,四甲基偶氮唑蓝试验(MTT)法检测细胞生长抑制情况。结果脂质体介导生存素反义寡核苷酸转染后的子宫内膜癌细胞出现生存素蛋白表达明显下降;ASODN转染组细胞凋亡率和增殖抑制率均明显高于各对照组(P<0.05),而各对照组间差异无统计学意义(P>0.05)。结论生存素反义寡核苷酸转染子宫内膜癌细胞能下调生存素蛋白表达,诱导子宫内膜癌细胞凋亡,抑制细胞增殖,对子宫内膜癌是一种有效的基因疗法。

Objective To investigate the effects of survivin antisense oligonucleotide （ASODN） on human uterine endometrial cancer HEC-1B cell line. Methods The experiment included four groups： blank control group （blank）, lipofectin control group （Lip）, oligonucleotide （SODN） control group and antisense oligonucleotide （ASODN） test group. Artificially synthesized survivin antisense oligonucleotide was transfected for 48 hours into endometrial cancer ceils using liposome after cultivation. The expression of survivin protein was determined by Western blot, and the cellular apoptotic rate was tested by flow cytometric analysis. MTT method was used to assay cellular growth rate. Results In comparing with the blank, Lip and SODN control groups, survivin protein expression was obviously decreased in HEC-1B cancer cells after transfected with survivin ASODN. The apoptotic rate and proliferation inhibition rate in ASODN test group were significantly higher than that in the control groups （P〈 0. 05）. There was no significant difference between other control groups （P〉0.05）. Conclusions The expression of survivin is decreased in endometrial cancer cells after ASODN transfection. Survivin ASODN causes cellular apoptosis and inhibits the proliferation of HEC-1B cells. It may be a useful gene therapy for endometrial cancer.