稳定表达Ras人成纤维细胞改变自噬活性细胞效应.PDFVIP

·1578 · doi 10.3969/j.issn. 1673-4254.2014.11.05 J South Med Univ, 2014, 34(11): 1578-1583 基础研究 稳定表达稳定表达RasRas 的人成纤维细胞改变自噬活性的细胞效应的人成纤维细胞改变自噬活性的细胞效应 1 1 2 2 王 玲 ，余 乐 ，古春萍 ，李亦蕾 1 2 南方医科大学 药学院，南方医院药学部，广东 广州 510515 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-RasV 12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，ATG7 siRNA 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。 关键词：自噬；Ras ；早期衰老；人成纤维细胞 Cellular response to altered autophagy activity in human fibroblast cells overexpressing Ha-RasV12 1 1 2 2 WANG Ling , YU Le , GU Chunping , LI Yilei 1 2 School of Pharmaceutical Science , Department of Pharmacology, Nanfang Hospital , Southern Medical University, Guangzhou 510515, China Abstract: 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 s