外源性p16基因导入对人膀胱癌细胞的作用

目的 通过外源性野生型Ｐ１６基因,纯翕生缺失及具有内源性Ｐ１６基因表达的膀胱癌细胞系２５３Ｊ和ＥＪ,地肿瘤细胞的生长抑制作用,并探讨共作用机制。方法 构建Ｐ１６逆转逆转录病毒表达载体,经细胞ＰＡ３４１７包装,膀胱癌细胞系;应用Ｎｏｒｔｈｅｍ杂交及免疫细胞化学方法检测外源１６基因的表达;流式我仪检测细胞周期;原位细胞凋亡检测及透射电镜观察细胞凋亡;并对导入的Ｐ１６基因的细胞进行鼠致瘤能力及病理不特性

Multiple inhibitory mechanisms of wild-type p16 gene transfer into human bladder cells by retroviral vector

Objective To investigate the inhibitory mechanisms of wild type p16 gene transfer into human bladder cancer cells with or without endogenous p16 gene expression. Methods p16 recombinant retrovirus vector was constructed and transfected in human bladder cancer cell EJ and 253J. Northern blot analysis and immunocytochemistry were used to detect the expression of exogenous p16 gene. Flow cytometry analysis was used to detect tumor cell cycle.Electronic microscope and in situ labelling apoptotic DNA fragment detection (TUNEL) was used to detect tumor cell apoptosis after gene transfection. Tumorigenicity was observed in nude mice. Results The growth rate of the transfected EJ cells was significantly retarded, the majority of transfected EJ cells were arrested on G 0 G 1 phases of cell cycle, tumorigenicity of the transfected cells was reduced on nude mice. H ras gene expression in transfected EJ cells reduced as compared with nontransfection.Apoptotic cells were observed in transgenic 253J cells. Conclusions Malignant proliferation of bladder cancer cells can be inhibited by exogenous p16 gene transfection via different molecular pathway.