【摘要】 目的研究抑制细胞周期蛋白依赖性激酶2(cyclin dependent kinase,CDK2)对人鼻咽癌细胞株CNE-2衰老的影响并探讨其分子机制。方法实验分为Control组、Control siRNA组、CDK2-siRNA组、CVT-313组,分别用PI单染流式细胞仪检测各组细胞增殖情况,衰老相关β-半乳糖苷酶染色法检测细胞衰老情况,流式细胞仪检测细胞活性氧(ROS)含量,Annexin-PI双染流式细胞仪检测细胞凋亡情况;进一步通过Western blot法检测衰老相关蛋白表达情况。结果人鼻咽癌细胞株CNE-2中,CDK2-siRNA组和CVT-313组细胞增殖指数明显降低,衰老细胞增多,活性氧含量增加,与对照组相比较差异具有统计学意义(P<0.05),而细胞凋亡率无显著性变化(P>0.05)。CDK2-siRNA组和CVT-313组p53、p21、p16蛋白表达明显增加,而pRb蛋白表达明显减少。结论抑制CDK2可诱导人鼻咽癌细胞株CNE-2的衰老,其机制可能是通过促进衰老相关蛋白的表达来实现的。更多还原

【Abstract】 OBJECTIVE To study the effects of cyclin-dependent protein kinase 2(CDK2) inhibition on senescence of human nasopharyngeal carcinoma cell line CNE-2 and to explore its molecular mechanism.METHODS The experiment was divided into control group,control siRNA group,CDK2-siRNA group and CVT-313 group.Cell proliferation was detected by PI monochrome flow cytometry,cell senescence was detected by aging-related beta-galactosidase staining,ROS content was detected by flow cytometry,and cell apoptosis was detected by Annexin-PI double-staining flow cytometry.Further,the aging-related protein expression was detected by Western blot.RESULTS In human nasopharyngeal carcinoma cell line CNE-2,the proliferation index of CDK2-siRNA group and CVT-313 group decreased significantly,the number of senile cells increased,and the content of ROS increased.Compared with the control group,the difference was statistically significant(P<0.05).While the apoptotic rate did not change significantly (P>0.05).The expression of p53,p21 and p16 protein in CDK2-siRNA group and CVT-313 group increased significantly.While the expression of pRb protein decreased significantly.CONCLUSION Inhibiting CDK2 can induce the senescence of human nasopharyngeal carcinoma cell line CNE-2,and the mechanism may be through promoting the expression of senescence-related proteins.更多还原