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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 effects of lupeol on the proliferation of human hepatoma Huh-7 cells. Methods: The effect of lupeol (10-150 μmol·L -1) on the viability of T24 cells was detected by CCK-8 assay. Caspase inhibitor was used to determine whether caspase-related protein was related to the inhibitory effect of lupeol on cell proliferation. The effect of lupeol on the mRNA expression of miR-21, MMP-2 and MMP-9 was evaluated by real-time PCR. The effect of lupeol on the expression of MMP-9 was analyzed by using the pGL3-Basic luciferase reporter system -9 promoter activity. Results: Lupeol inhibited the proliferation of Huh-7 cells at a dose-dependent manner at 72 h. The IC50 of lupeol-treated cells at 72 h was 80 μmol·L -1. Compared with the control group, Compared with the control group, lupeol can reduce the mRNA expression of MMP-9 and decrease the expression of MMP-9 in Huh-7 cells compared with the control group The gene promoter activity was down-regulated by 40%. The difference between the above results and the control group was statistically significant (P <0.05). Conclusion: Lupeol can inhibit the proliferation of human hepatoma Huh-7 cells, which is related to the inhibition of miR-21 / c-Jun / MMP-9 signaling pathway.