韩丽萍, 刘娟芳, 赵国强, 董子明. RNA干扰靶向沉默poIβ基因对人卵巢癌细胞增殖活性及对化疗药物敏感性的影响[J]. 中国肿瘤临床, 2011, 38(4): 197-199 . DOI: 10.3969/j.issn.1000-8179.2011.04.005
引用本文: 韩丽萍, 刘娟芳, 赵国强, 董子明. RNA干扰靶向沉默poIβ基因对人卵巢癌细胞增殖活性及对化疗药物敏感性的影响[J]. 中国肿瘤临床, 2011, 38(4): 197-199 . DOI: 10.3969/j.issn.1000-8179.2011.04.005

RNA干扰靶向沉默poIβ基因对人卵巢癌细胞增殖活性及对化疗药物敏感性的影响

  • 摘要: 目的:探讨RNA干扰(RNA interference, RNAi)沉默DNA聚合酶β(polβ)基因的表达后,人卵巢癌细胞HO-8910的增殖活性变化及对化疗药物顺铂敏感性的改变。方法: 利用已构建成功的针对polβ基因的siRNA (small interfering RNA,siRNA)重组质粒,转染人卵巢癌细胞HO-8910,通过荧光倒置显微镜观察转染效率; 荧光定量PCR检测转染后HO-8910细胞中polβmRNA的表达水平;用MTT法观察检测基因沉默后HO-8910细胞增殖能力变化;台盼蓝染色细胞计数法了解细胞对顺铂敏感性的变化。结果:已构建的靶向polβ基因的siRNA转染人卵巢癌细胞HO-8910后,荧光倒置显微镜下可见大量绿色荧光颗粒表达;荧光定量PCR检测发现,转染后细胞中polβ的mRNA水平表达显著降低(P<0.01);转染后的HO-8910细胞生长速度减慢、增殖活性下降;对顺铂的敏感性提高(P<0.05)。结论:RNAi有效沉默细胞中polβ基因表达,提高了肿瘤细胞对化疗药物顺铂的敏感性、降低了细胞的增殖活性。从而改善卵巢癌患者的预后,为肿瘤的有效干预和治疗开拓新的领域。

     

    Abstract: Effect of polβ siRNA on Proliferation of Human Ovarian Carcinoma Cell Line and ItsSensitivity to CisplatinLiping HAN1, Juanfang LIU1, Guoqiang ZHAO2, Ziming DONG3Corresponding author: Liping HAN, E-mail: hanliping0825@yahoo.com.cn1Department of Gynecology and Obstetrics, The First Hospital of Zhengzhou University, Zhengzhou 450052, China2Department of Immunology and Microbiology, Medical College of Zhengzhou University, Zhengzhou 450001, China3Department of Pathophysiology, Medical College of Zhengzhou University, Zhengzhou 450001, ChinaAbstract Objective: To investigate the effect of siRNA of polβ on the proliferation of human carcinoma cell (HO-8910) and itssensitivity to cisplatin. Methods: The plasmid expressing siRNA against polβ was constructed and transfected into HO-8910 cells. Thetransfection efficiency was evaluated by fluorescence microscopy. pol β expression was detected by Fluorogenic Quantitative PCR.MTT was used to observe the proliferation of HO-8910 cells and Trypan blue dyeing was used to determine the sensitivity of HO-8910cells to cisplatin. Results: After successful transfection of the constructed siRNA -pol β gene into HO-8910, a large number offluorescent particles were observed in the fluorescence inverted microscope. The expression of pol β in mRNA was significantlyreduced ( P < 0.01 ). The ability of growth and proliferation were reduced. On the other hand, the sensitivity to cisplatin was enhanced( P < 0.05 ). Conclusion: polβ siRNA in ovarian carcinoma cells could induce proliferation of HO-8910 cell line but can enhance itssensitivity to cisplatin.Keywords polβ; siRNA; HO-8910; Cisplatin; Fluorogenic quantitative PCR

     

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