腺病毒介导的p16基因表达对体外细胞增殖和衰老的影响  

Impact of adenovirus-mediated p16 expression on cell proliferation and senescence in vitro

作　　者：吴红平[1] 孙娟娟[1] 李林芳[1] 钱其军[1] 

机构地区：[1]第二军医大学东方肝胆外科医院病毒及基因治疗实验室,上海200438

出　　处：《中国癌症防治杂志》2015年第5期320-324,共5页CHINESE JOURNAL OF ONCOLOGY PREVENTION AND TREATMENT

基　　金：国家科技重大专项项目(2013ZX10002010-007)

摘　　要：目的探讨p16基因表达对正常细胞和肿瘤细胞增殖和衰老的影响。方法采用腺病毒载体构建并包装过表达p16基因的腺病毒，分别感染原代小鼠成纤维细胞（MEF）和肝癌SMMC-7721细胞，用CCK8法检测细胞增殖，SA-β-gal染色检测细胞衰老。结果成功构建腺病毒介导的p16基因表达系统，并在MEF和SMMC-7721细胞中高效表达。CCK8法检测p16基因表达对MEF细胞增殖的抑制率在第1～4天分别为（6．8±0．25）％、（10．6±0．68）％、（12．4±0．93）％和（45．7±1．13）％（P〈0．01）；但对SMMC-7721细胞增殖抑制不明；SA-β-gal染色显示p16基因可明显诱导MEF细胞衰老[（15±5）个/视野]，但不能诱导SMMC-7721细胞发生衰老（0个/视野）。结论单独p16基因表达在体外不足以诱导肿瘤细胞的衰老和生长阻滞。Objective To investigate the influence of exogenous p16 expression on cell proliferation and senescence in normal and tumor cells. Method Genes encoding p16 and GFP were subctoned into an adenovirus vector, then recombinant adenovirus was produced in 293 cells and purified for subsequent infection into MEF and SMMC7721 cells. Cell proliferation was examined by CCK8 assay and senescence by SA-β-gal staining. Results The number of senescent MEF cells was much higher in euhures after infection with adenovirus expressing p16 than after infection with control adenovirus （15±5 vs 0 senescent cells/field）, and pl6-expressing cultures showed significantly lower proliferation on days 1-4[ （6.8±0.25）%, （ 10.6±0.68 ）%, （ 12.4±0.93 ）% and （45.7±1.13）%;P〈0.01 ]. In contrast, no significant differences in proliferation or senescence were observed between control SMMC-7721 cultures or cultures expressing p16. Conclusion Expression of p16 alone is insufficient to induce cell growth arrest and senescence in tumor cells.

关 键 词：肿瘤 P16基因 衰老 增殖 腺病毒 

分 类 号：R730.2[医药卫生—肿瘤]