CAJ | 학술논문

目的观察雷帕霉素联合应用表阿霉素抑制HepG2细胞的程度,并探讨自噬在其中的作用.方法分组用药后,测定对HepG2细胞增殖的抑制作用；流式细胞仪检测细胞线粒体膜电位,荧光染色法检测自噬体形成以及Western blot测定p62、Beclin1、人微管相关蛋白1轻链3(LC3)Ⅰ、LC3Ⅱ的蛋白表达水平.结果联合用药后,抑瘤率(67.00±2.45)％较单药组(23.00±1.85)％显著增加(P＜0.05),线粒体膜电位(荧光比61.14％)较单药组(荧光比25.73％)明显下降(P＜0.01),自噬囊泡增多,并明显增加了LC3Ⅱ/LC3 Ⅰ比值以及p62表达.结论雷帕霉素阻断雷帕霉素靶蛋白(mTOR)通路可诱导HepG2细胞自噬,联合表阿霉素时能增加自噬水平,提高HepG2细胞的化疗敏感性.
목적 관찰뢰파매소연합응용표아매소억제HepG2세포적정도,병탐토자서재기중적작용.방법 분조용약후,측정대HepG2세포증식적억제작용；류식세포의검측세포선립체막전위,형광염색법검측자서체형성이급Western blot측정p62、Beclin1、인미관상관단백1경련3(LC3)Ⅰ、LC3Ⅱ적단백표체수평.결과 연합용약후,억류솔(67.00±2.45)％교단약조(23.00±1.85)％현저증가(P＜0.05),선립체막전위(형광비61.14％)교단약조(형광비25.73％)명현하강(P＜0.01),자서낭포증다,병명현증가료LC3Ⅱ/LC3 Ⅰ비치이급p62표체.결론 뢰파매소조단뢰파매소파단백(mTOR)통로가유도HepG2세포자서,연합표아매소시능증가자서수평,제고HepG2세포적화료민감성.
Objective To explore the effects of rapamycin combined with epirubicin on proliferation of HepG2 cells and the roles of autophagy.Methods HepG2 cells were given rapamycin combined with epirubicin or alone,and divided into four groups.The inhibitory effects of rapamycin combined with epirubicin or alone on proliferation of HepG2 cells were assessed.Mitochondrial membrane was tested by using flow cytometry.The autophagic bodies were observed by using fluorescence staining,and the protein expression levels of p62,LC-3 Ⅰ,LC3 Ⅱ and Beclin1 were detected by using Western blotting.Results The viability of HepG2 cells in rapamycin + epimbicin group was increased significantly as compared with single group [(67.00 ± 2.456) ％ vs.(23.00 ± 1.85) ％,P ＜ 0.05].Mitochondrial membrane in rapamycin + epirubicin group was decreased significantly as compared with single group [green fluorescence ratio (61.14％) vs.(25.73％),P ＜ 0.05].In rapamycin + epirubicin group,the expression of p62 and LC3 Ⅱ was up-regulated.Conclusion Blocking the mammalian target of rapamycin (mTOR) pathway can induce death of cancer cells through autohagic mechanisms,and combined use of rapamycin and epirubicin may synergically enhance the effects of activating autophagy.