小分子干扰RNA下调星形细胞上调基因1的表达对卵巢癌细胞增殖和凋亡的影响

Effect of down-regulation of astrocyte elevated gene-1 expression by short interfering RNA on cell proliferation and apoptosis in ovarian cancer cell lines

目的探讨小分子干扰RNA(siRNA)下调星形细胞上调基因1(AEG-1)表达对卵巢癌细胞增殖和凋亡的影响。方法卵巢癌细胞株SKOV3分为3组:空白对照组(不做任何处理)、阴性对照组(对照siRNA转染)和观察组(AEG-1 siRNA转染)。用反转录聚合酶链反应观察AEG-1 siRNA对AEG-1基因表达的影响;甲基噻唑基四唑法观察AEG-1 siRNA对卵巢癌SKOV3细胞增殖的抑制作用;流式细胞术检测AEG-1 siRNA对卵巢癌SKOV3细胞凋亡和细胞周期的影响。结果空白对照组和阴性对照组AEG-1 mRNA表达、SKOV3细胞增殖、细胞凋亡率及在DNA合成前期(G0/G1期)、合成期(S期)、合成后期(G2/M期)的细胞比例差异均无统计学意义(P>0.05)。观察组SKOV3细胞AEG-1 mRNA表达显著低于空白对照组和阴性对照组,差异有统计学意义(P<0.01)。观察组SKOV3细胞增殖在24、48、72 h较空白对照组和阴性对照组均显著减慢,差异有统计学意义(P<0.01)。观察组SKOV3细胞凋亡率显著高于空白对照组和阴性对照组,差异有统计学意义(P<0.01);观察组SKOV3细胞在DNA合成前期(G0/G1期)细胞比例显著高于空白对照组和阴性对照组(P<0.01),而合成期(S期)、合成后期(G2/M期)的细胞比例均显著低于空白对照组和阴性对照组(P<0.01),差异均有统计学意义。结论 siRNA可通过下调卵巢癌SKOV3细胞AEG-1基因的表达,抑制其增殖,促进其凋亡。

Objective To investigate the effect of short interfering RNA（siRNA）-induced astrocyte elevated gene-1（AEG-1） down-regulation on cell proliferation,apoptosis and cell cycle of ovarian cancer cells.Methods SKOV3 cells were divided into blank control group（without treatment）,negative control group（control siRNA transfection） and observation group（AEG-1 siRNA transfection）.The influence of AEG-1 siRNA on expression of AEG-1 mRNA was observed by reverse transcription-polymerase chain reaction（RT-PCR）.The cell proliferation was detected by methylthiazolyldiphenyl-tetrazolium bromide method.The apoptosis and cell cycle were analyzed by flow cytometry.Results There was no significant difference between blank control group and negative group in the expression of AEG-1 mRNA,SKOV3 cell proliferation,the rate of cell apoptosis and the cell percentage in DNA G0/G1 phase,S phase and G2/M phrase.Compared with those in blank control group and negative control group,the expression of AEG-1 mRNA was evidently declined in the observation group（P〈 0.01）.SKOV3 cell proliferation in observation group were significantly slower at 24,48 and 72 h compared with those in blank control group and negative control group（P〈 0.01）.After treated with AEG-1 siRNA for 72 h,the percentage of apoptotic cells increased significantly compared with those in the blank control group and negative control group（P〈 0.01）.The cell percentage in DNA G0/G1 phase of observation group was significantly higher than those in blank group and negative group,while the cell percentage in S phase and G2/M phase of observation group were significantly lower than those in blank group and negative group.Conclusion AEG-1 siRNA in ovarian cancer cells can significantly inhibit cell proliferation and promote apoptosis through down-regulating the expression of AEG-1 in SKOV3 cells.