文章摘要
刘 瑾,吴锦昌,周俊东.POLⅠ基因对人食管癌细胞放化疗敏感性的影响[J].南京医科大学学报,2013,(1):6~10
POLⅠ基因对人食管癌细胞放化疗敏感性的影响
Effects of POLⅠ on radiosensitivity of human esophageal cancer cell line and its sensitivity to cisplatin
投稿时间:2012-09-18  
DOI:10.7655/NYDXBNS20130102
中文关键词: POLⅠ  shRNA  ECA109  放疗敏感  顺铂
英文关键词: POLⅠ  shRNA  ECA-109  radiosensitivity  cisplatin
基金项目:苏州市科技局社会发展基金(SYS201254)
作者单位
刘 瑾 南京医科大学附属苏州医院放疗科,江苏 苏州 215001 
吴锦昌 南京医科大学附属苏州医院放疗科,江苏 苏州 215001 
周俊东 南京医科大学附属苏州医院放疗科,江苏 苏州 215001 
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中文摘要:
      目的:探讨DNA聚合酶Ⅰ(DNA polymeraseⅠ,POLⅠ)基因对人食管癌细胞ECA-109放射敏感性及对化疗药物顺铂敏感性的影响。方法:利用已经构建成功的针对POLⅠ基因的短发夹RNA(short hairpin RNA,shRNA)重组质粒,转染ECA-109细胞,通过倒置荧光显微镜观察转染效率,实时荧光定量PCR检测转染后ECA-109细胞中POLⅠ的mRNA表达水平,用细胞克隆形成实验来研究shRNA抑制POLⅠ表达后ECA-109细胞对X线放疗敏感性的变化,用MTT法检测POLⅠ基因敲除后ECA-109细胞对顺铂敏感性的变化。结果:沉默POLⅠ基因的shRNA转染ECA-109细胞后,荧光倒置显微镜下可见大量绿色荧光颗粒;实时荧光定量PCR检测发现,转染后细胞中POLⅠ的mRNA表达水平显著降低(P < 0.05);转染后的细胞对放疗敏感性提高,对顺铂的敏感性提高(P < 0.05)。结论:shRNA有效下调细胞中POLⅠ的表达,提高了肿瘤细胞对化疗药物顺铂和放疗的敏感性。
英文摘要:
      Objective:To investigate the effect of POLⅠ (DNA polymeraseⅠ)on the radiosensitivity of human esophageal cancer cells (ECA-109)and their sensitivity to cisplatin. Methods:The plasmid expressing shRNA against POLⅠ was constructed and transfected into ECA-109 cells. The transfection efficiency was evaluated by fluorescence microscopy. POLⅠ expression was detected by Real-time Fluorogenic Quantitative PCR. The cell sensitivity to X-ray was detected by colony formation assay. MTT was used to determine the sensitivity of ECA-109 cells to cisplatin. Results:After successful transfection of constructed shRNA-POLⅠ gene into ECA-109 cells,a large number of fluorescent particles were observed in fluorescence inverted microscope. The expression of POLⅠ in mRNA was significantly reduced (P < 0.05). The ability of radiosensitivity was enhanced. On the other hand,sensitivity to cisplatin was enhanced (P < 0.05). Conclusion:ShRNA-POL I in human esophageal cancer cells could enhance their radiosensitivity and sensitivity to cisplatin.
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