外源性P16基因导入对人膀胱癌细胞增殖及凋亡的影响

Effects of transfer of P16 gene on the proliferation and apoptosis of human bladder cancer cell

目的 :将外源性P16基因导入该基因纯合性缺失的人膀胱癌细胞系 ,观察对肿瘤细胞增殖活性及凋亡的影响。方法 :构建P16逆转录病毒表达载体 ,转染膀胱癌细胞系 ;应用Northern杂交及免疫细胞化学检测外源性P16基因的表达 ;流式细胞仪检测细胞周期 ;原位细胞凋亡检测及透射电镜观察细胞凋亡。结果 :转染外源P16基因的肿瘤细胞生长速率明显降低 ,Ki6 7标记指数下降 ,大多数细胞被抑制在G0 和G1期 ,同时发现凋亡细胞的存在。结论 :外源性P16基因导入不仅抑制膀胱癌细胞的增殖 ,且可诱导其凋亡。

Aim: To observe the effects of p16 gene transfer on the proliferation and apoptosis of human bladder cancer cell without endogenous p16 gene expression. Methods: p16 recombinant retrovirus vector was contructed and transfected in human bladder cancer cell 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) were used to detect tumor cell apoptosis after gene transfection. Results: The growth rate of the transfected cells was significantly retarded, Ki67 labelling index was significantly decreased. The majority of transfected cells was arrested at G 0+G 1 phases of cell cycle. Apoptotic cells were found in transgenic 253J cells. Conclusion: p16 gene would play an important role in the regulation of the imbalance between the proliferation and apoptosis of human bladder cancer cell.