p16基因高表达对293细胞衰老的影响  

Effects of p16 gene overexpression on 293 cells aging

作　　者：马珊珊[1] 刘静[1] 姚宁[1] 崔渊博[1] 邢衢 王梦然[2] 郭天宇[3] 杨波[4] 关方霞[1,4] 

机构地区：[1]郑州大学生命科学学院干细胞实验室,郑州450001 [2]华东师范大学生命科学学院,上海200241 [3]中山大学生命科学学院,广州510275 [4]郑州大学第一附属医院,郑州450052

出　　处：《郑州大学学报（医学版）》2014年第5期622-625,共4页Journal of Zhengzhou University(Medical Sciences)

基　　金：国家自然科学基金资助项目81071008;河南省杰出青年基金114100510005;河南省科技攻关项目122102310203;人事部留学回国人员择优资助项目

摘　　要：目的:探讨p16基因高表达对293细胞衰老的影响。方法:实验设正常组、空质粒组和高表达组。RTPCR扩增人p16基因,构建p16基因高表达载体,利用脂质体将其转染293细胞。采用CCK-8法和流式细胞术检测p16基因高表达对293细胞增殖和细胞周期的影响,β-半乳糖苷酶染色法检测细胞衰老状态,RT-PCR和Western blot法检测p16、p21 mRNA和蛋白的表达。结果:p16基因高表达可抑制293细胞的增殖,并将细胞阻滞在G0/G1期(F=158.057,P<0.001);高表达组细胞p16和p21在mRNA和蛋白水平的表达明显增加(F=73.467、54.988、9.923、44.060,P均<0.05),β-半乳糖苷酶阳性细胞率明显增加(F=137.266,P<0.001)。结论:p16基因高表达可抑制293细胞的增殖,将细胞阻滞在G0/G1期,提高细胞中p16和p21的表达,促进细胞衰老。Aim：To investigate the regulation of overexpression of p 16 gene on aging of 293 cells.Methods：The ex-periment had 3 groups：control group,empty plasmid group,and overexpression group.The p16 gene was amplified by RT-PCR to construct the p16 gene overexpression vector ,and the vector was transfected into 293 cells.CCK-8 assay and flow cytometry were performed to detect the effect of p 16 gene overexpression on cell proliferation and cell cycle .β-galactosidase staining was used to detect the cell senescence state ,and the expressions of p 16 and p21 in mRNA and protein level were analyzed by RT-PCR and Western blot .Results：p16 gene overexpression could inhibit the proliferation of 293 cells and the cell cycle was arrested in G0/G1 phase（F=158.057,P〈0.001）.The expressions of p16 and p21 in the overexpression group increased in the mRNA and protein level （F=73.467,54.988,9.923,44.060,P〈0.05）,and the percentage of cells staining positively for β-galactosidase increased （F=137.266,P 〈0.001）.Conclusion：p16 gene overexpression could inhibit the proliferation of 293 cells,arrest the cell in G0/G1 phase,improve the expression of p16 and p21,and pro-mote the aging .

关 键 词：P16 高表达 细胞衰老 调控 293细胞 

分 类 号：Q291[生物学—细胞生物学]