CAJ | 학술논문

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，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。
목적：연구인성섬유세포은정과도표체암기인Ras대자서적영향。방법이자서억제제록규、자서관건기인ATG7적siRNA、자서촉진제뢰파매소처리전염료H-RasV12혹공재체적BJ세포，관찰기대세포증식、쇠로화사망지표적영향。결과록규처리후，Ras은정과도표체적BJ세포쇠로현상경가명현차세포사망솔현저승고，ATG7 siRNA역가이촉진세포쇠로，이뢰파매소처리후사망솔동양현저승고，단존활세포적쇠로감경。재대조세포，록규촉진세포쇠로화사망적세포효응발생지완，ATG7 siRNA화뢰파매소처리칙무명현작용。결론인성섬유세포은정과도표체암기인Ras후，자서활성수도억제차억제정도수도엄격적공제，개변자서활성가대세포쇠로화존활산생현저영향。
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.