siRNA沉默Bmi-1基因表达对宫颈癌细胞增殖的影响研究
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Effect of siRNA silencing Bmi-1 gene on cell proliferation of cervical cancer cells
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    摘要:

    目的 探讨siRNA沉默B细胞特异的莫洛尼白血病病毒插入位点1基因(Bmi-1)表达对宫颈癌细胞增殖的影响研究。方法 通过脂质体转染siRNA Bmi-1及阴性对照(control)到人宫颈癌Hela细胞,采用Realtime PCR及western blot法检测细胞中Bmi-1的表达。MTT法检测细胞活力,Annexin V-PI流式双染及Hoechst染色检测细胞凋亡情况,流式细胞术检测细胞周期,western blot法检测细胞中p16,p53,CyclinD1及Rb的表达。结果 与Control组比较,siRNA Bmi-1组细胞中Bmi-1蛋白及mRNA表达量皆显著降低(P<0.01),同时细胞活力下降,细胞凋亡率提高(P<0.01),细胞周期阻滞在G1期(P<0.01),细胞中p16表达量上调(P<0.01),p53及CyclinD1表达量下调(P<0.01),Rb磷酸化水平降低(P<0.01)。结论 Bmi-1具有抑制宫颈癌Hela细胞增殖的作用,可能与调控细胞周期相关蛋白表达有关。

    Abstract:

    Objective To explore effect of siRNA silencing B-cell specific Moloney murine leukemiavirus integration site (Bmi-1) gene expression on cell proliferation of cervical cancer cells. Methods Hela cells were transferred with control and siRNA Bmi-1. The expression of Bmi-1 in Hela cell was examed by Realtime PCR and western blot. The cell viability was detected by MTT. The cell apoptosis was examed by Annexin V-PI flow cytometry and Hoechst staining. Cell cycle was detected by flow cytometry. The expression of p16, p53, CyclinD1 and Rb was detected by western blot. Results Compared with control group, the expression of Bmi-1 protein and mRNA was down-regulated, the cell viability was decreased, cell apoptotic rate was increased, Cell cycle was arrested in the G1 phase, the expression of p16 was up-regulated, the expresson of p53 and CyclinD1 down-regulated, the phosphorylation of Rb was inhibited in siRNA Bmi-1 group. The difference was statistically significant (P<0.01). Conclusion Bmi-1 inhibited Hela cell proliferation via regulation of expression of cell cycle related protein.

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张冬丽. siRNA沉默Bmi-1基因表达对宫颈癌细胞增殖的影响研究[J].中国比较医学杂志,,().

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  • 收稿日期:2016-05-23
  • 最后修改日期:2016-06-16
  • 录用日期:2016-06-28
  • 在线发布日期: 2016-11-28
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