作　　者： (）;

机构地区： 广东医科大学研究生学院

出　　处： 《中华实验外科杂志》 2018年第10期1855-1858,共4页

摘　　要： 目的观察JIB-04对肝癌细胞增殖、凋亡的影响，探讨其影响发生的机制。方法用不同浓度的组蛋白去甲基化酶抑制剂JIB-04（0、4、8、16μmol／L）分别处理肝癌细胞HepG2、QGY7703、Huh7、MHCC-97H、MHCC-97L及正常肝细胞L02，采用细胞计数试剂盒（CCK-8）检测细胞增殖变化；流式胞仪检测细胞肝癌细胞HepG2、Huh7及MHCC-97H细胞周期分布及凋亡；选取肝癌细胞HepG2及Huh7行蛋白质印迹法检测凋亡蛋白表达。结果JIB-04对正常肝细胞L02抑制作用不明显，经最高浓度为16μmol／L培养48h后．抑制率仅为6％，而不同浓度JIB-04对各肝癌细胞分别有不同程度的抑制作用，其中对肝癌细胞Huh7及QGY7703抑制作用最明显，其半数抑制浓度（IC50）分别为3、3．5μmol／L，且JIB-04对各肝癌细胞抑制作用呈现时间及浓度依赖性；在流式细胞仪检测中，各肝癌细胞S、M期比例明显减少，而G，期比例明显升高且呈药物浓度依赖性；同时随着JIB-04浓度的增加，肝癌细胞HepG2、Huh7及MHCC-97H凋亡率明显提高，经最高浓度8μmol／L作用48h后其凋亡率分别为（27．84±0．42）％、（25．31±0．93）％、（46．86±0．63）％，且不同浓度组与对照组两两比较其凋亡率差异有统计学意义；在凋亡蛋白检测中，肝癌细胞凋亡蛋白B细胞淋巴瘤／白血病-2相关X蛋白（bax）均上调及B细胞淋巴瘤／白血病-2（bcl-2）均下调，两者肝癌细胞中凋亡蛋白p53均明显上调。结论JIB-04对正常肝细胞抑制作用不明显，但对不同肝癌细胞有不同程度的抑制作用；JIBJD4通过干扰细胞周期分布及诱导凋亡来抑制肝癌细胞增殖；JIB-04有可能通过线粒体凋亡途径来诱导肝癌细胞凋亡。 Objective To study the effect of JIB - 04 on the proliferation and apoptosis of hepatocellular carcinoma （HCC） cells and the mechanism. Methods JIB - 04, histone demethylase inhibitor, with different concentrations （0, 4, 8, 16 μmol/L） were used to treat hepatoma cells HepG2, QGY7701, QGY7703, Huh7, MHCC -97H, MHCC -97L and normal hepatocytes L02. The cell counting kit -8 （ CCK - 8 ） kit was used to detect cell proliferation of different hepatoma cells. The flow cytometry was used to detect cell cycle arrest and cell death apoptosis of HepG2, Huh7 and MHCC -97H cells. Hepatoma cells HepG2 and Huh7 were selected to detect the expression of apoptotic proteins by Western blotting. Results JIB -04 had no obvious inhibitory effect on H）2 cells. After treatment with 16 μmol/L JIB -04 for 48 h, the inhibition rate was only 6% and different concentrations of JIB -04 showed different inhibitory effects on each liver cancer cell line. The inhibitory effect of JIB - 04 on Huh7 and QGY7703 ceils were strongest, with the half maximal inhibitory concentration （ICs0） being 3 and 3.5 μmol/L, respectively, and the inhibitory effect of JIB - 04 on each liver cancer cell line was time - and concentration - dependent. The flow cytometry revealed that the proportion of S phase and M phase of HepG2, Huh7 and MHCC -97H HCC cells was significantly reduced, and that of Gt phase was significantly increased. At the same time, with the increase of JIB -04 concentration, the apoptosis rate of HepG2, Huh7 and MHCC -97H cells was sig- nificantly increased, Which was respectively （ 27.84 ± 0. 42 ） %, （ 25.31 ± 0. 93 ） % and （46. 86 ±0.63）% after treatment with JIB -04 8 μmol/L for 48 h. There was statistically significant difference be-tween control group and different concentration JIB - 04 groups. The expression of protein B cell lymphoma/ leukemia- 2 associated X protein （bax） and p53 increased, and that of B cell lymphoma/leukemia -2 （bcl- 2） was downregulated in hepG2 and MHCC -97H cells. Conclu

关 键 词： 癌 肝细胞 脱噬作用 增殖

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