摘要
目的通过腺病毒携带p16基因感染胃癌细胞,研究p16功能恢复对CDK4表达的调节作用。方法构建携带p16基因的重组腺病毒AdCMV-p16感染胃癌细胞系。Western blotting检测p16和CDK4的表达,MTT法检测癌细胞的增殖活性,DAPI染色计数癌细胞的凋亡比例。结果胃癌细胞感染腺病毒载体AdCMV-p16后获得p16表达,对细胞整体CDK4表达无影响,但可明显降低细胞核CDK4的表达量;AdCMV-p16感染后引起癌细胞增殖活性下降,当感染复数(MOI)为1、10、20时,细胞存活率已经分别低于50%、20%和5%;p16表达可诱导癌细胞凋亡,细胞凋亡率达(13.86±4.65)%。结论 p16功能恢复后核CDK4含量减少,可能是诱导细胞周期阻滞和细胞凋亡、抑制癌细胞生长的主要分子机制。
Objective To investigate the regulation of CDK4 expression by reactivating p16 function through adenovirus vector in gastric cancer.Methods The adenovirus vector carrying p16 gene of AdCMV-p16 was constructed and used to infect gastric cancer cell lines.Western blotting was used to detect the expression of p16 and CDK4,MTT assay was used to detect the proliferation activity of cancer cells,and DAPI was used to stain and count the percentages of cancer cell apoptosis.Results After reactivation of p16 in gastric cancer cells,the expression level of CDK4 in whole cell extracts was not affected,but the nuclear CDK4 was decreased obviously.This phenomenon resulted in the depression of cancer cell proliferation activity,with the cell viability of lower than 50%,20%,or 5% when MOI was 1,10 or 20,respectively.p16 expression induced gastric cancer cells apoptosis with the apoptotic rate of（13.86±4.65）%.Conclusion The reactivation of p16 function in gastric cancer cells resulted in the decrease of nuclear CDK4,which may be the main molecular mechanism of p16-induced cell cycle arrest and p16-mediated inhibition of cancer cell proliferation.
出处
《临床肿瘤学杂志》
CAS
2011年第2期102-104,共3页
Chinese Clinical Oncology
基金
浙江省科技计划资助项目(2006C30021)
