前列腺癌特异突变DNA聚合酶β表达载体转染NIH3T3的细胞生物学行为变化  

Effect of expression vector containing mutant DNA polβ gene derived from prostate cancer on cellular biological behavior when transfected into NIH3T3 cell

作　　者：王明臣[1] 霍玉奇[1] 张善锋[1] 宋宇[1] 赵勤[1] 金国平[1] 董子明[2] 

机构地区：[1]郑州大学基础医学院生物化学与分子生物学教研室,河南郑州450052 [2]郑州大学基础医学院病理学与病理生理学教研室

出　　处：《中国老年学杂志》2010年第16期2292-2295,共4页Chinese Journal of Gerontology

基　　金：河南省科技厅自然科学基金(0611043400)

摘　　要：目的观察前列腺癌(CPa)特异突变DNA聚合酶β(polβ)表达载体转染NIH3T3的细胞生物学行为变化。方法采用业已成功构建的2例特异突变polβ表达载体(pCDNA3.1-M1和pCDNA3.1-M2),采用脂质体包裹转染NIH3T3细胞,观察比较这些突变对细胞polβ基因mRNA和蛋白表达的影响以及引发的相应的细胞周期和细胞自发突变率等细胞生物学行为的变化。结果筛选得到高表达PCa特异突变polβ的NIH3T3细胞株(NIH3T3-M1,NIH3T3-M2)和高表达野生型polβ的NIH3T3细胞株(NIH3T3-W)。RT-PCR和Western印迹结果显示,外源DNApolβ在转化的NIH3T3细胞中呈现高表达。与转染空载体和未转染的NIH3T3细胞相比,转染PCa特异突变polβ的2个细胞株和转染野生型polβ的细胞株的细胞周期的S期比例及转染细胞的HGPRT(次黄嘌呤-鸟嘌呤磷酸核糖转移酶)基因的自发突变率均显著增加,且转染野生型polβ的细胞株亦显著高于转染PCa特异突变DNApolβ的细胞株。结论 DNApolβ高表达介导了肿瘤发生的分子机制。Objective To observe the effect of expression vector containing mutant DNA polβ gene derived from prostate cancer on cellular biological behavior when they were transfected into NIH3T3 cell.Methods Two eukaryotic expression vectors containing mutant DNA polβ derived from prostate cancer （pcDNA3.1-M2 and pcDNA3.1-M2） were transfected into NIH3T3 cell lines and the effects on gene expression of DNA polβ and the corresponding cellular biological behavior including the changes of cell cycle and cellular spontaneous mutation rate were observed.Results The NIH3T3 cell line which overexpressed mutant DNA polβ gene derived from prostate cancer and the NIH3T3 cell line which overexpressed wild-type polβ gene were all obtained and also presented.The cellular biological behavior included the increase of S-period frequency （SPF） of cell cycle and the elevation of spontaneous mutation rate of HGPRT gene.Moreover,both the SPF of cell cycle and spontaneous mutation rate of HGPRT gene in cells transfected with wild-type DNA polβ were significantly higher than those in the cells transfected with specific mutant DNA polβ derived from prostate cancer.Conclusions Tumor genesis is due to the overexpression of DNA polβ.

关 键 词：前列腺癌 DNA聚合酶Β 表达载体 NIH3T3 细胞周期 自发突变率 

分 类 号：R735[医药卫生—肿瘤]