稳定表达Ras的人成纤维细胞改变自噬活性的细胞效应  

Cellular response to altered autophagy activity in human fibroblast cells overexpressing Ha-RasV12

作　　者：王玲[1] 余乐[1] 古春萍[2] 李亦蕾[2] 

机构地区：[1]南方医科大学药学院,广东广州510515 [2]南方医科大学南方医院药学部,广东广州510515

出　　处：《南方医科大学学报》2014年第11期1578-1583,共6页Journal of Southern Medical University

基　　金：国家自然科学基金(81102475);南方医科大学南方医院高层次课题匹配基金(81102475)~~

摘　　要：目的研究人成纤维细胞稳定过度表达癌基因Ras对自噬的影响。方法以自噬抑制剂氯喹、自噬关键基因ATG7的siRNA、自噬促进剂雷帕霉素处理转染了H-RasV12或空载体的BJ细胞,观察其对细胞增殖、衰老和死亡指标的影响。结果氯喹处理后,Ras稳定过度表达的BJ细胞衰老现象更加明显且细胞死亡率显著升高,ATG7 siRNA亦可以促进细胞衰老,而雷帕霉素处理后死亡率同样显著升高,但存活细胞的衰老减轻。在对照细胞,氯喹促进细胞衰老和死亡的细胞效应发生迟缓,ATG7siRNA和雷帕霉素处理则无明显作用。结论人成纤维细胞稳定过度表达癌基因Ras后,自噬活性受到抑制且抑制程度受到严格的控制,改变自噬活性可对细胞衰老和存活产生显著影响。Objective To study the effect of oncogenic Ras overexpression on autophagic activity in human fibroblast cells in vitro. Methods BJ cells were transfected with H-RasV12 or control vector and treated with chloroquine, small interfering RNA （siRNA） for ATG7, or rapamycin. The cellular responses were analyzed by monitoring the parameters and biomarkers for cell growth, senescence and cell death. Results In BJ cells overexpressing H-RasV12, chloroquine treatment resulted in more prominent cell senescence and a significantly increased cell death rate. Suppression of ATG7 mediated by siRNA also promoted cell senescence. Rapamycin treatment also caused an increased cell death rate but attenuated senescence in surviving cells. In control BJ cells, the cellular response to chloroquine included senescence and cell death, which occurred slowly. Rapamycin treatment and siRNA suppression of ATG7 had no obvious effect on control BJ cells. Conclussion Stable cellular overexpression of oncogenic Ras causes tightly controlled suppression of the autophagic activity of human fibroblast cells, and such changes produce significant effect on cell senescence and survival.

关 键 词：自噬 RAS 早期衰老 人成纤维细胞 

分 类 号：Q25[生物学—细胞生物学]