海峡药学
海峽藥學
해협약학
Strait Pharmaceutical Journal
2015年
10期
256-258
,共3页
Torin1%细胞周期%cyclin%A1%P27
Torin1%細胞週期%cyclin%A1%P27
Torin1%세포주기%cyclin%A1%P27
Torin1%cellcycle%cyclin A1%P27
目的 研究Torin1 对人肝癌HepG2 细胞生长的影响及其可能机制. 方法 CCK8 法检测不同浓度(0.1,0.5,1.25,2.5,5)μmol· L -1 Torin1 对HepG2细胞48h后细胞增殖的影响;流式细胞仪检测不同浓度(1.25,2.5,5)μmol· L -1 Torin1 处理前后HepG2 细胞周期的变化,Real time PCR和Western blot法检测不同浓度(1.25,2.5,5)μmol· L -1 Torin1处理前后HepG2细胞细胞周期抑制蛋白P27 和细胞周期蛋白A1(cy-clinA1) mRNA表达水平和蛋白表达水平的变化. 结果 Torin1 作用于 HepG2 细胞后,HepG2 细胞增殖明显呈浓度依赖型抑制,1.25,2.5, 5μmol· L -1 Torin1作用于HepG2细胞48 h后,细胞周期S期明显增多,P27 mRNA和蛋白表达水平呈浓度依赖性增加,cyclinA1 mRNA和蛋白表达水平呈浓度依赖性减少. 结论 体外应用Torin1可显著抑制人肝癌细胞HepG2 细胞生长,这可能是通过增加抑制细胞周期蛋白 P27 表达,减少细胞周期蛋白A1的表达来阻碍细胞周期进程.
目的 研究Torin1 對人肝癌HepG2 細胞生長的影響及其可能機製. 方法 CCK8 法檢測不同濃度(0.1,0.5,1.25,2.5,5)μmol· L -1 Torin1 對HepG2細胞48h後細胞增殖的影響;流式細胞儀檢測不同濃度(1.25,2.5,5)μmol· L -1 Torin1 處理前後HepG2 細胞週期的變化,Real time PCR和Western blot法檢測不同濃度(1.25,2.5,5)μmol· L -1 Torin1處理前後HepG2細胞細胞週期抑製蛋白P27 和細胞週期蛋白A1(cy-clinA1) mRNA錶達水平和蛋白錶達水平的變化. 結果 Torin1 作用于 HepG2 細胞後,HepG2 細胞增殖明顯呈濃度依賴型抑製,1.25,2.5, 5μmol· L -1 Torin1作用于HepG2細胞48 h後,細胞週期S期明顯增多,P27 mRNA和蛋白錶達水平呈濃度依賴性增加,cyclinA1 mRNA和蛋白錶達水平呈濃度依賴性減少. 結論 體外應用Torin1可顯著抑製人肝癌細胞HepG2 細胞生長,這可能是通過增加抑製細胞週期蛋白 P27 錶達,減少細胞週期蛋白A1的錶達來阻礙細胞週期進程.
목적 연구Torin1 대인간암HepG2 세포생장적영향급기가능궤제. 방법 CCK8 법검측불동농도(0.1,0.5,1.25,2.5,5)μmol· L -1 Torin1 대HepG2세포48h후세포증식적영향;류식세포의검측불동농도(1.25,2.5,5)μmol· L -1 Torin1 처리전후HepG2 세포주기적변화,Real time PCR화Western blot법검측불동농도(1.25,2.5,5)μmol· L -1 Torin1처리전후HepG2세포세포주기억제단백P27 화세포주기단백A1(cy-clinA1) mRNA표체수평화단백표체수평적변화. 결과 Torin1 작용우 HepG2 세포후,HepG2 세포증식명현정농도의뢰형억제,1.25,2.5, 5μmol· L -1 Torin1작용우HepG2세포48 h후,세포주기S기명현증다,P27 mRNA화단백표체수평정농도의뢰성증가,cyclinA1 mRNA화단백표체수평정농도의뢰성감소. 결론 체외응용Torin1가현저억제인간암세포HepG2 세포생장,저가능시통과증가억제세포주기단백 P27 표체,감소세포주기단백A1적표체래조애세포주기진정.
OBJECTIVE To investigate in vetro the effect of Torin1 on the proliferation and cell cycle on HepG2 cell line and explore their molecular mechanism.METHODS The anti-proliferative effect on HepG2 cells was determined by CCK8 assay.The cell cycle was measured by flow cytometry.The Cyclin A1 ,P27 expression levels of HepG2 cells treated with Torin1 were detected in mRNA and protein by Real time PCR and Western blot.RESULTS Torin1 could inhibit cell proliferation on HepG2 cells,and the inhibitory effects were in dose de-pendent manners.Cell cycle analysis showed that the S phase of HepG2 cells treated with Torin1 increased signifi-cantly compared with the control group.Real time quantitative PCR and Western blot results showed that Torin1 could down-regulate Cyclin A1 expressions ( P <0.05 ) and up-regulate p27 expression ( P <0.05 ) .CONCLUSION Torin1 has the anti-proliferative effect on HepG2 cells and induce S phase arrest.These effects might be related with proteins associated with cell cycle closely,such as Cyclin A1 and P27.
