Apoptin对宫颈癌Hela细胞放疗增敏作用实验研究

摘要

目的 探讨Apoptin增加宫颈癌Hela细胞的敏感性及作用机制.方法 应用脂质体转染方法将表达Apoptin的重组载体pEGFP-Apoptin转染入人宫颈癌Hela细胞.其中转染pEGFP-Apopti为实验组,转染pEGFP-C2组为阳性对照组,未转染组为阴性对照组.上述各组经射线干预后利用平板克隆形成实验、流式细胞术、MTT及蛋白印迹western-blot等方法分别检测细胞凋亡、周期及相关蛋白表达水平.结果 pEGFP-Apoptin在人宫颈癌Hela细胞中稳定表达.Apoptin可增加宫颈癌Hela细胞对放射线的敏感性,Apoptin可使p21、p27的表达量上调,使Rad50及Ku80的表达量下调,而对XLF的表达量则无影响.结论 Apoptin可增加宫颈癌Hela细胞的放疗敏感性,其机制可能与细胞周期阻滞及同源与非同源重组修复相关蛋白的表达水平改变有关.%Objective To observe whether the cervical carcinoma Hela cells have increased the radiation sensitivity by radiotherapy after apoptin transfection into the above cells.Further,its possible mechanism is explored.Methods The cervical carcinoma Hela cells were transfectd by the recombinant vectors encoding a GFP-Apoptin (pEGFP-Apoptin).One day after the viral infection,cells concurrent exposure to irradiation.Cells survive were detected by the MTT assay and plate clone formation assay.The cells cycle were detected by FCM (flow cytometry).The expressions of proteins were detected by Western Blot.Results PEGFP-Apoptin was effective in cervical carcinoma Hela cells.Apoptin enhances radiation-induced cell death in cervical carcinoma Hela cells.The regulatory proteins expression in the cells cycle of the Apoptin transfection team,such as p21,p27,was significantly increased.The DNA damage repair proteins,Rad50,Ku80 inApoptin transfection team were significantly lower than one in the GFP transfection term and negative term.XLF molecule protein has no significantly changed.Conclusion The combination therapy with radiation and apoptin may increase the sensitivity to radiation for cervical carcinoma Hela cells.The mechanism may be associated with cell cycle relative proteins and DNA repair protein.