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目的:研究母源性肿瘤抑制印迹基因ARHI对人卵巢癌SKOV3细胞株的作用。方法:将pIRES2-EGFP-ARHI质粒转染至ARHI基因低表达的人卵巢癌SKOV3细胞内实现ARHI基因表达,CCK-8法检测pIRES2-EGFP-ARHI-SKOV3细胞组(实验组)、pIRES2-EGFP-SKOV3细胞组(质粒对照组)及SKOV3细胞组(阴性对照组)的吸光度值,计算细胞生长抑制率,流式细胞仪分析细胞周期分布并检测细胞凋亡率,并用Western blot检测微管相关蛋白轻链Ⅱ(LC3-Ⅱ)的表达水平变化。结果:实验组细胞培养24、48、72、96及120 h的生长抑制率分别为64.69%、70.17%、67.01%、66.87%、67.70%,均明显高于质粒对照组(P﹤0.01)。培养48 h时实验组、质粒对照组、阴性对照组S期细胞的比例分别为64.18%、38.43%及15.15%,凋亡率分别为47.97%、26.53%及9.33%;培养72 h时S期细胞的比例分别为43.29%、10.37%及10.89%,凋亡率分别为51.34%、24.70%及4.39%;实验组细胞培养48 h时LC3-Ⅱ表达水平增加,与对照组间有明显差异。结论:ARHI基因抑制SKOV3细胞生长,使SKOV3细胞阻滞于S期,并诱导凋亡及自体吞噬。
Objective: To study the effect of maternally derived tumor suppressor gene ARHI on human ovarian cancer SKOV3 cell line. Methods: ARHI gene was transfected into SKOV3 cells with low expression of ARHI gene. The expression of ARHI gene in pIRES2-EGFP-ARHI-SKOV3 cells (experimental group) and pIRES2-EGFP The cell growth inhibition rate was calculated by the absorbance value of the cells in the SKOV3 cell group (control group) and the SKOV3 cell group (negative control group). The cell cycle distribution was analyzed by flow cytometry and the apoptosis rate was detected. Protein light chain Ⅱ (LC3-Ⅱ) expression levels. Results: The growth inhibition rates of experimental group at 24, 48, 72, 96 and 120 h were 64.69%, 70.17%, 67.01%, 66.87% and 67.70%, respectively, which were significantly higher than those of plasmid control group (P <0.01). The percentage of S phase cells in experimental group, plasmid control group and negative control group were 64.18%, 38.43% and 15.15% respectively at the 48th hour of culture. The apoptotic rates were 47.97%, 26.53% and 9.33% The cell apoptosis rate was 43.29%, 10.37% and 10.89%, respectively. The apoptotic rates were 51.34%, 24.70% and 4.39% respectively. The expression of LC3-Ⅱ in experimental group increased at 48 h, which was significantly different from that in control group. Conclusion: The ARHI gene inhibits the growth of SKOV3 cells, arrests SKOV3 cells in S phase and induces apoptosis and autophagy.