摘要
目的 观察细胞周期素依赖性激酶-8（CDK8）基因的稳定沉默对SW480增殖及生长的影响,并探讨其机制.方法 慢病毒感染SW480后筛选出单克隆；荧光定量聚合酶链反应（FQ-PCR）检测基因表达；噻唑蓝（MTT）比色法和裸鼠成瘤检测细胞的增殖和移植瘤的生长；流式细胞术分析细胞周期和凋亡.结果 CDK8基因沉默组细胞培养4d后的生长抑制率为NC对照组的39.6％,裸鼠移植瘤的体积比NC对照组减小86.8％；CDK8基因沉默组细胞的凋亡率和G0/G1期细胞百分率分别为6.55％和56.46％；显著高于NC对照组（P＜0.05）.结论 CDK8基因稳定沉默能显著抑制SW480细胞的增殖和移植瘤的生长,细胞凋亡及细胞周期阻滞与CDK8沉默效应有关.
Objective To investigate the effect of stable cyclin dependent kinase inhibitor8 （CDK8） knockdown on growth of sw480 cells in vitro and in vivo,and to illuminate the mechanisms.Methods Single cell clone of SW480 was obtained by puromycine screen after lentivirus infection.mRNA expression was determined by using quantitative reverse transcriptase-polymerase chain reaction.Cell proliferation and tumor growth were analyzed by methyl thiazol tetrazolium （MTT） assay and tumor formation in nude mice.Cell cycle and apoptosis were detected by using flow cytometry.Results Compared to normal control group,the inhibition rate of SW480 clone at day 4 was 39.6% and the tumor volume of transplanted tumor at day 25 was decreased by 86.8%.The apoptosis rate and proportion of G0/G1 phase of SW480 clone were 6.55% and 56.46% respectively,which was significantly increased as compared with normal control group.Conclusion Stable CDK8 knockdown inhibited the proliferation of SW480 cells and tumor growth in naked mices,promoted SW480 cells apoptosis and arrested these cells in the G0/G1 phase.
出处
《中华实验外科杂志》
CAS
CSCD
北大核心
2013年第11期2308-2310,共3页
Chinese Journal of Experimental Surgery
基金
广东省科技计划资助项目（2012B031800025）
广东省自然科学基金资助项目（06301124）
