ARHI基因诱导人卵巢癌SKOV3细胞株S期阻滞、凋亡及自体吞噬的研究
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南京市医学科技发展基金项目(YKK10037);南京医科大学科技发展基金项目(09NJMUM101)


ARHI gene induces S phase arrest, apoptosis and autophagy of human ovarian cancer SKOV3 cell lines
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    摘要:

    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。

    Abstract:

    Objective:To study the effects of aplasia rashomolog memberⅠ(ARHI ) on HOC SKOV3 cell lines. Methods:After transfection of the pIRES2-EGFP-ARHI plasmid to the HOC SKOV3 cells with low expression level of ARHI gene, absorbance A was measured and performed to calculate the inhibitory rate of cell growth using CCK-8 method. To analyze the cell cycle distribution and apoptosis rate by using flow cytometry and to test the change in expression level of LC3-Ⅱ protein by using Western blot approach in the pIRES2-EGFP-ARHI-SKOV3 (the test group), pIRES2-EGFP-SKOV3 (the plasmid control group) and SKOV3 (the negative control group) cell groups. Results:After the cells in the test group were cultured for 24, 48, 72, 96 and 120 h, we found that the inhibitory rates of cell growth were 64.69%, 70.17%, 67.01%, 66.87% and 67.70%, respectively. The inhibitory rates of cell growth in the test group were significantly higher than those in the plasmid control group(P < 0.01). The ratios of S phase cells were 64.18%, 38.43% and 15.15%, and the apoptosis rates were 47.97%, 26.53% and 9.33% respectively after culturing the cells for 48 h in the test, plasmid control and negative control group. The proportions of cells in S phase were 43.29%, 10.37% and 10.89%, and the apoptosis rates were 51.34%, 24.70% and 4.39% respectively after culturing the cells in the test, plasmid control and negative control group for 72 h. The significant difference remained between the test and control groups. The expression level of LC3-Ⅱ in test group increased with significant difference compared with the control group after cultured for 48 h. Conclusion:These findings suggest that ARHI gene inhibits the growth of SKOV3 cells, arrests the SKOV3 cells at S phase and induces apoptosis and autophagy.

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朱巧英,孟惠娟,周金华,沈宇飞,胡建铭. ARHI基因诱导人卵巢癌SKOV3细胞株S期阻滞、凋亡及自体吞噬的研究[J].南京医科大学学报(自然科学版),2013,(7):902-906

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  • 收稿日期:2013-03-03
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  • 在线发布日期: 2013-07-09
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