摘要
目的观察过氧化物酶体增殖物激活受体γ(PPARγ)的配体罗格列酮(RSG)在体外对人肝癌细胞增殖、凋亡的影响及其作用机制。方法应用CCK-8比色法检测不同浓度(25、50、100、200μmol/L)RSG对肝癌HepG2细胞增殖的影响;应用Hoechst33258荧光染色观察细胞凋亡形态学变化;流式细胞仪检测细胞凋亡率及分析细胞周期;实时荧光定量PCR法观察Bcl-2、Bax的mRNA表达;免疫细胞化学法检测Bcl-2、Bax蛋白的表达。结果RSG对肝癌HepG2细胞的增殖抑制率与浓度呈正相关。肝癌HepG2细胞的凋亡率与RSG浓度呈正相关;与对照组比较,加药组S期细胞百分率降低,G1期细胞百分率升高(P均<0.05)。100、200μmol/L RSG组的凋亡细胞较对照组明显增加,且浓度越高增加越显著。RSG干预肝癌HepG2细胞后,Bcl-2的mRNA及蛋白表达在肝癌细胞中下调,而Bax的mRNA及蛋白在肝癌细胞中表达上调。结论RSG可通过激活PPARγ、调节Bcl-2和Bax的水平而在体外抑制肝癌细胞生长,并诱导其凋亡。
Objective To investigate the effect of peroxisome proliferators activated receptor gamma （PPARγ） ligand rosiglitazone （RSG） on proliferation and apoptosis in human hepatocarcinoma cell. Methods Detected the effect of RSG in different concentrations （25, 50, 100,200 umol/L）and different action times （24, 48, 72 hours）on the growth of hepatocareinoma cells HepG2 by Cell Counting Kit-8 （CCK-8） analysis. The change of cell apoptosis rate and the cell cycle in each concentration were detected by flow cytometry （ FCM ）. Detected the morphological change of apoptosis by Hoechst 33258. The expression of Bcl-2 and Bax mRNA and protein were investigated by real-time fluorescent quantitive polymerase chain reaction and immunocytochemieal method respectively. Results RSG could inhibit the proliferation of HepG2 cells in concentration dependent and time dependent which were in positive correlation. Compared with control group, the apoptosis rates of RSG （100, 200 umol/L）were significantly enhanced. Cell cycle was arrested at G1 stage by RSG in a concentration dependent manner （ P 〈 0.05 ）. Hoeehst 33258 fluorescence staining confirmed that the apoptosis body was located in the HepG2 cell treating with RSG. Real-time Flurescent Quantitive Polymerase Chain Reaction and Immunohistochemical techniques indicated the down-regulation of Bcl-2 and the up-regulation of Bax in mRNA and protein expression. Conclusion These results indicates that the growth-inhibitory and apoptosis effect of hepatocarcinoma cell in vitro is mediated through PPARγ signaling pathway by RSG, suggesting that PPARγ might be a new therapeutic target for hepatocarcinoma.
出处
《山东医药》
CAS
北大核心
2009年第1期10-13,共4页
Shandong Medical Journal
关键词
过氧化物酶体增殖物激活受体Γ
罗格列酮
肝肿瘤
增殖
凋亡
peroxisome proliferator activated receptor gamma
rosiglitazone
liver neoplasms
proliferation
apoptosis
