熊果酸调控miR-134对人肝癌细胞增殖和侵袭的影响  

Ursolic acid inhibits the proliferation and invasion of human hepatoma cells by regulating miR-134

作　　者：陈诚 曹梦蝶 鲁晓雨 盛丽婷 虞燕霞[1] CHEN Cheng;CAO Mengdie;LU Xiaoyu;SHENG Liting;YU Yanxia(Clinical Trials Institution,Suzhou Municipal Hospital,Affiliated Hospital of Nanjing Medical University,Jiangsu Suzhou 215002,China)

机构地区：[1]南京医科大学附属苏州医院/苏州市立医院临床试验机构,江苏苏州215002

出　　处：《中国医院药学杂志》2023年第17期1939-1942,1954,共5页Chinese Journal of Hospital Pharmacy

基　　金：苏州市科技发展计划项目(编号:SYSD2019207)。

摘　　要：目的:探讨熊果酸通过调控miR-134对人肝癌细胞增殖和侵袭的影响。方法:体外培养人肝癌细胞株SMMC-7721,CCK-8法和Transwell实验检测不同浓度的熊果酸对SMMC-7721细胞增殖和侵袭的影响;Western blot检测上皮标志物E-cadherin蛋白、间质标志物N-cadherin和Vimentin蛋白的表达和转染miR-134模拟物后整合素β1(integrin β1,ITGB1)蛋白表达水平的差异;双荧光素酶报告基因确定miR-134与ITGB1 3’-UTR的靶向结合情况;RT-qPCR法检测用0~30μmol·L-1熊果酸处理后SMMC-7721细胞中miR-134的表达量。结果:熊果酸对肝癌细胞的生长抑制有剂量依赖性影响,80μmol·L-1时最为明显(P<0.01);熊果酸能升高上皮标志物E-cadherin蛋白在SMMC-7721细胞中的表达(P<0.01),降低间质标志物N-cadherin和Vimentin蛋白的表达(P<0.01);熊果酸能够剂量依赖性促进肝癌细胞中miR-134的表达(P<0.01);ITGB1是miR-134的直接靶基因,过表达miR-134能够显著抑制ITGB1蛋白的表达(P<0.01);转染miR-134模拟物后SMMC-7721细胞中ITGB1蛋白的表达显著降低(P<0.01)。结论:熊果酸诱导的miR-134通过减少SMMC-7721细胞中ITGB1的表达显著抑制EMT进程,从而抑制细胞增殖和侵袭。OBJECTIVE To explore the effects of ursolic acid on the proliferation and invasion of human HCC cells by regulating miR-134.METHODS Human hepatoma cell line SMMC-7721 was cultured in vitro.CCK-8 method and Transwell assay were respectively used to detect the proliferation and invasion of SMMC-7721 cells treated with different concentrations of ursolic acid.The expression of E-cadherin,N-cadherin and Vimentin,and the expression of integrinβ1(ITGB1)after transfection with miR-134 mimics were measured by Western blot.Dual-luciferase reporter assay was conducted to test the binding between miR-134 and ITGB13'-UTR.RT-qPCR was used to detect the expression of miR-134 in SMMC-7721 cells treated with 0-30μmol·L-1 ursolic acid.RESULTS Ursolic acid inhibited the growth of hepatoma cells in a dose-dependent manner.80μmol·L-1 Ursolic acid had the most obvious inhibitory effect in hepatoma cells(P<0.01).Ursolic acid could up-regulate the expression of epithelial marker E-cadherin protein in SMMC-7721 cells(P<0.01),and down-regulate the expression of N-cadherin and Vimentin(P<0.01).Ursolic acid promoted the expression of miR-134 in hepatoma cells in a dose-dependent manner(P<0.01).ITGB1 was a direct target gene of miR-134.Overexpression of miR-134 could significantly inhibit the expression of ITGB1 protein(P<0.01).After transfection with miR-134 mimics,the expression of ITGB1 protein in SMMC-7721 cells decreased significantly(P<0.01).CONCLUSION Ursolic acid significantly inhibits EMT procession by reducing ITGB1 expression through inducing miR-134 in SMMC-7721 cells,thereby suppressing the cell proliferation and invasion.

关 键 词：熊果酸 肝细胞癌 miR-134 ITGB1 增殖 侵袭 

分 类 号：R285.5[医药卫生—中药学]