转染NK4基因对膀胱癌移行上皮细胞HGF基因表达的影响  

Introduction of exogenous NK4 gene into bladder transitional epithelial cells can reduce the HGF in tumor cells

作　　者：潘卫兵[1] 张青汉[1] 余毅恺[2] 叶绪龙[1] 董能本[1] 彭伟[1] Toshikazu Nakamura 

机构地区：[1]湖北省黄石市中心医院泌尿外科,435000 [2]华中科技大学同济医学院免疫学部 [3]日本长崎大学病理学部

出　　处：《中华实验外科杂志》2007年第5期576-577,共2页Chinese Journal of Experimental Surgery

基　　金：湖北省科技攻关计划资助项目(6AA301C46)

摘　　要：目的观察转染NK4基因对膀胱癌上皮细胞HGF基因表达的影响。方法利用脂质体Lipofectin-2000将质粒pcDNA3(+)/NK4导入BIU-87膀胱癌细胞,用pcDNA3(+)作为阴性对照。G418稳定筛选阳性细胞克隆后,逆转录-聚合酶链反应(RT-PCR)检测转染细胞和对照组细胞NK4基因mRNA水平表达,Western blot检测转染后细胞培养上清液NK4和细胞胞质HGF蛋白的表达含量。结果转染阳性细胞NK4 mRNA恒定表达,RT-PCR和Western blot分别显示在mR- NA转录水平和蛋白表达水平,稳定转染的膀胱癌细胞NK4含量明显高于对照组。转染了NK4基因的肿瘤细胞HGF蛋白表达水平明显降低,而转染空载体则无作用。结论脂质体将外源NK4基因导入肿瘤细胞后能稳定表达NK4蛋白,而且可以有效降低肿瘤细胞HGF表达的水平。Objective To study the influence of NK4 transfection on bladder tumor transitional epithelial cells and to provide theoretical basis for gene therapy of bladder tumor. Methods peDNA3 （ ＋ ）/NK4 was introduced into BIU-87 bladder tumor cell lines with the Lipofeetin-2000, and plasmid eDNA3（ ＋ ） as the positive control. The positive clone was selected with G418. NK4 mRNA was measured in transfeeted cells and the control group eeUs by RT-PCR and Western-blot was applied to detect the NK4 and HGF expression. Results Positively transfeeted cells stably expressed NK4 mRNA. RT-PCR and Western-blot revealed the stably transfeeted bladder tumor eeUs. Conclusion Tumor cells transfeeted with exogenous NK4 gene can stably express NK4 protein and can effectively reduce the HGF in tumor cells.

关 键 词：膀胱肿瘤 基因治疗 

分 类 号：R737.14[医药卫生—肿瘤]