Retroviral vector containing human p16 gene and its inhibitory effect on Bcap-37 breast cancer cells  

含人p16基因逆转录病毒载体的构建及其对Bcap-37乳腺癌细胞的抑制作用(英文)

作　　者：白景香[1] 祝学光[1] 郑新程[2] 伍贻经[2] 

机构地区：[1]北京医科大学人民医院外科,北京100044 [2]北京医科大学病理生理教研室,北京100083

出　　处：《Chinese Medical Journal》2001年第5期49-53,106,共6页中华医学杂志（英文版）

摘　　要：Objective To study the effects of the p16 gene on tumor cell growth inhibition and cell cycle arrest. Methods The recombinant retroviral vector pLp16SN was constructed by cloning the human p16 cDNA into the retroviral vector pLXSN. Retroviruses with or without the p16 gene were obtained by transfecting pLp16SN and pLXSN vectors into PA317 cells. Bcap-37 human breast cancer cells were infected with these retroviruses followed by selection with G418. The expression of p16 was detected by Northern and Westem blots. Cell biological characteristics, including cell growth rate, cell cycle and tumorigenesis in nude mice were assessed.Results Both mRNA and protein expression of p16 in Bcap-37 cells transfected with retroviral vector containing the p16 gene were much higher than that in Bcap-37 cells transfected with empty vector or parental Bcap-37 cells. Cell overexpressing the p16 gene exhibited a slower rate of growth, a higher percentage of cells in the G1 phase, and smaller tumors in nude mice, compared with parental Bcap-37 cells and cells transfected with empty vector.Conclusion Overexpression of the p16 gene could suppress the growth of Bcap-37 breast cancer cells by arresting the cell cvcle at the G1 to S-phase.目的　研究p16基因对肿瘤细胞生长抑制及细胞周期阻滞作用。方法　将p16cDNA插入逆转录病毒载体pLXSN构建成p16基因逆转录病毒重组体pLp16SN。利用基因转染方法 ,将pLp16SN及pLXSN导入逆转录病毒包装细胞系PA317细胞 ,获得逆转录病毒。用逆转录病毒感染Bcap 37乳腺癌细胞 ,经G4 18筛选获阳性克隆。利用Northern和Westernblotting方法检测p16基因的表达。检测转基因细胞的生长速度 ,细胞周期及裸鼠成瘤等细胞生物学行为的改变。结果　Northern及Westernblotting显示转染p16基因的Bcap 37细胞p16基因mRNA及蛋白质表达明显增高。此细胞较未转染基因的Bcap 37细胞和转染空载体的Bcap 37细胞生长速度慢 ,G1期细胞比率增高 ,裸鼠接种成瘤体积小。结论　p16基因高表达能够抑制乳腺癌细胞Bcap 37的生长 。

关 键 词：p16 gene  ·   retroviral vector   ·  breast cancer cells ·  cell cycle arrest 

分 类 号：R737.9[医药卫生—肿瘤]