CAJ | 학술논문

目的探讨二甲双胍(Met)对肝癌细胞衰老的影响及其机制.方法不同浓度(0、0.01、0.1、1、10和50 mmol/L) Met处理肝癌细胞HepG2细胞后,采用MTS法检测细胞增殖情况;流式细胞术检测细胞周期及细胞凋亡改变;通过衰老相关β-半乳糖苷酶(SA-β-ga1)染色和衰老标记分子Dec1蛋白表达分析Met对细胞衰老的影响;蛋白质印迹法分析p-AMPK、p-ACC和AMPK蛋白的表达.结果 Met可以抑制HepG2细胞的增殖,且呈浓度依赖性;高浓度(10和50 mmol/L)Met促进肝癌细胞凋亡;低浓度(0.01、0.1和1 mmol/L) Met作用后,肝癌细胞呈现典型的大而扁平的衰老形态,SA-β-ga1染色阳性细胞显著增多,细胞周期停滞于G0/G1期,衰老标记分子Dec1蛋白表达明显上调.此外,低浓度Met可以促进p-AMPK和p-ACC蛋白表达,而对AMPK蛋白表达无明显影响.结论高浓度Met促进肝癌细胞凋亡;低浓度Met则诱导肝癌细胞衰老,其作用机制可能与激活AMPK信号通路有关.该研究为以后利用诱导肝癌细胞衰老这种方式提高肝癌综合治疗水平,提供了重要实验依据.
목적 탐토이갑쌍고(Met)대간암세포쇠로적영향급기궤제.방법 불동농도(0、0.01、0.1、1、10화50 mmol/L) Met처리간암세포HepG2세포후,채용MTS법검측세포증식정황;류식세포술검측세포주기급세포조망개변;통과쇠로상관β-반유당감매(SA-β-ga1)염색화쇠로표기분자Dec1단백표체분석Met대세포쇠로적영향;단백질인적법분석p-AMPK、p-ACC화AMPK단백적표체.결과 Met가이억제HepG2세포적증식,차정농도의뢰성;고농도(10화50 mmol/L)Met촉진간암세포조망;저농도(0.01、0.1화1 mmol/L) Met작용후,간암세포정현전형적대이편평적쇠로형태,SA-β-ga1염색양성세포현저증다,세포주기정체우G0/G1기,쇠로표기분자Dec1단백표체명현상조.차외,저농도Met가이촉진p-AMPK화p-ACC단백표체,이대AMPK단백표체무명현영향.결론 고농도Met촉진간암세포조망;저농도Met칙유도간암세포쇠로,기작용궤제가능여격활AMPK신호통로유관.해연구위이후이용유도간암세포쇠로저충방식제고간암종합치료수평,제공료중요실험의거.
Objective To explore the effect of metformin on hepatoma cells senescence and the underlying mechanism.Methods Cell proliferation,cycle and apoptosis were examined by MTS and flow cytometry assay in response to different concentrations of metformin(0,0.01,0.1,1,10 and 50mmol/L).Senescence-associated β-galactosidase (SA-β-ga1) staining and senescence marker Dec1 protein levels were used to evaluate the effect of metformin on hepatoma cells senescence.In addition,protein expression of p-AMPK,p-ACC and AMPK was detected by Western blot analysis.Results Metformin suppressed proliferation of HepG2 cells in a dose-dependent manner.High concentrations of metformin (10 and 50mmol/L) promoted cell apoptosis,while lower doses of metformin (0.01,0.1 and 1mmol/L) led to enlarged and flatten senescent morphology and increased SA-β-ga1 positive cells.Moreover,cell cycle was blocked in G0/G1 phase and protein levels of senescent marker Dec1,p-AMPK and p-ACC were significantly enhanced,whereas AMPK protein expression was almost unchanged.Conclusion We showed here that high dose of metformin promotes HepG2 cells apoptosis,but low doses of metformin induce cellular senescence,which may be related to the activation of AMPK signaling.These data will provide vital evidence for improving the outcome of comprehensive treatment in HCC patients by driving hepatoma cells to undergo senescence.