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。
Objective： To investigate the effect of siRNA of polβ on the proliferation of human carcinoma cell （HO-8910） and its sensitivity to cisplatin. Methods： The plasmid expressing siRNA against polβ was constructed and transfected into HO-8910 cells. The transfection efficiency was evaluated by fluorescence microscopy, polβ 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-8910 cells to cisplatin. Results： After successful transfection of the constructed siRNA-polβ gene into HO-8910, a large number of fluorescent particles were observed in the fluorescence inverted microscope. The expression of polβ in mRNA was significantly reduced （ 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 its sensitivity to cisplatin.
出处
《中国肿瘤临床》
CAS
CSCD
北大核心
2011年第4期197-199,共3页
Chinese Journal of Clinical Oncology
