羽扇豆醇抑制人肝癌Huh-7细胞增殖机制研究  

Anti-proliferative Mechanism of Lupeol on Human Hepatocellular Carcinoma Huh-7 Cells

作　　者：唐泽严 杨小彤 周旭 邱振鹏 Tang Zeyan Yang Xiaotong Zhou Xu Qiu Zhenpeng(College of Pharmacy, Hubei University of Chinese Medicine, Wuhan 430065, Chin)

机构地区：[1]湖北中医药大学药学院,武汉430065

出　　处：《中国药师》2017年第9期1521-1525,共5页China Pharmacist

基　　金：国家自然科学基金项目(编号:81603338)

摘　　要：目的:研究羽扇豆醇(lupeol)对人肝癌Huh-7细胞增殖的影响。方法:运用CCK-8法检测lupeol(10~150μmol·L^(-1))对T24细胞活力的影响;运用caspase抑制剂确证凋亡相关蛋白caspase是否与lupeol对细胞增殖抑制作用有关;运用western blot检测lupeol对c-Jun蛋白表达的影响;运用real-time PCR评价lupeol对miR-21、MMP-2、MMP-9基因mRNA表达的影响;运用pGL3-Basic荧光素酶报告系统研究lupeol对MMP-9启动子活性的影响。结果:Lupeol在72 h能够抑制Huh-7细胞增殖,该抑制作用具有剂量依赖性;lupeol处理细胞72 h后半抑制浓度(IC50)为80μmol·L^(-1);与对照组相比,运用lupeol能够使Huh-7细胞中c-Jun蛋白表达下调;lupeol能够降低miR-21表达水平;与对照组相比,lupeol处理Huh-7细胞降低MMP-9的mRNA表达,并使MMP-9基因启动子活性下调40%;以上结果与对照组比较差异均有统计学意义(P<0.05)。结论:Lupeol能够抑制人肝癌Huh-7细胞增殖,该作用与抑制miR-21/c-Jun/MMP-9信号通路有关。Objective： To study the anti-proliferative effect of lupeol on human hepatocellular carcinoma Huh-7 cells. Methods： CCK-8 assay was performed to evaluate the effects of lupeol on cell viability. Caspase inhibitors were used to confirm whether caspase family was involved in lupeol-induced cell death. The effects of lupeol on the expression of c-Jun protein, miR-21 and mRNA of MMP- 9 were evaluated by western blot and real-time PCR, respectively, pGL3-Basic luciferase reporter assay system was used to study the effects of lupeol on MMP-9 promoter activity. Results： Lupeol could inhibit the proliferation of Huh-7 ceils in 72 h,and the inhibitory effect was dose-dependent. The half-inhibitory concentration （ IC50 ） of lupeol treated ceils was 80μmol·L^-1. Lupeol could down - regulate the expressions of c-Jun and miR-21 when compared with the control group. The mRNA expression of c-Jun targets and MMP- 9 was suppressed by lupeol. Moreover, the transcriptional activity of MMP-9 was down-regulated to 40% while the cells were treated with lupeol. Compared with the control group, the differences were statistically significant （P 〈0.05）. Conclusion： The results indicated that lupeol can inhibit Huh-7 cell proliferation, and the effects are involved in inhibiting miR-21/c-Jun/MMP-9 cascade.

关 键 词：羽扇豆醇 肝细胞癌 细胞增殖 

分 类 号：R965.1[医药卫生—药理学]