黄芩素下调微小RNA-130a抑制肝癌细胞侵袭机制研究

Study on mechanism of baicalein down-regulating miR-130a to inhibit hepatocellular carcinoma invasion

目的:探讨黄芩素通过下调微小RNA-130a(miR-130a)抑制肝癌细胞侵袭的机制。方法:以肝癌细胞HepG2为研究对象,应用慢病毒转染法上调HepG2细胞miR-130a表达并设为miR-130a组,转染空载慢病毒作为空载对照组,以未转染细胞作为空白组。采用实时荧光定量PCR检测miR-130a表达水平;黄芩素处理miR-130a组细胞;侵袭小室实验考察细胞的侵袭能力;蛋白印迹实验检测基质金属蛋白酶-9(MMP-9)、钙蛋白酶抑制蛋白(Calpastatin)表达水平。结果:与空载对照组相比,miR-130a组细胞miR-130a表达明显上调,细胞侵袭率升高,MMP-9蛋白表达上调,Calpastatin蛋白表达下降,差异有统计学意义(均P<0.05)。与miR-130a组相比,黄芩素加miR-130a组的miR-130a、MMP-9蛋白表达均降低,Calpastatin蛋白表达升高,细胞侵袭率显著降低,差异有统计学意义(均P<0.01)。结论:miR-130a通过CANP-MMP-9信号通路促进HepG2细胞侵袭,黄芩素能抑制miR-130a诱导的CANP-MMP-9通路激活,抑制HepG2细胞侵袭。

Objective:To explore mechanism of baicalein regulating miR-130a expression and inhibiting invasion of liver cancer cells.Methods:Hepatocarcinoma cell HepG2 was used as the research object,and the miR-130a expression of HepG2 cells was up-regulated by lentiviral transfection and set as the miR-130a group.The transfected empty-loaded lentivirus was used as the non-load control group,and the untransfected cells were used as the blank group.The expression level of miR-130a was detected by real-time fluorescent quantitative PCR,and the cells of the miR-130a group were treated with baicalein.Invasion chamber experiment was used to examine the invasion ability of cells,and western blotting experiment was used to detect the expression levels of matrix metalloproteinase-9(MMP-9)and Calpastatin.Results:Compared with the no-load control group,the expression of miR-130a in the miR-130a group was significantly up-regulated,cell invasion rate increased,the expression of MMP-9 protein up-regulated,and the expression of Calpastatin protein down-regulated,differences statistically significant(all P<0.05).Compared with the miR-130a group,the expression of miR-130a and MMP-9 protein were significantly down-regulated in the baicalein+miR-130a group,Calpastatin protein expression up-regulated,and cell invasion rate reduced,differences statistically significant(all P<0.01).Conclusion:miR-130a can promote HepG2 cell invasion through CANP-MMP-9 signaling pathway.Baicalein can inhibit miR-130a-induced activation of CANP-MMP-9 pathway,thereby inhibiting HepG2 cell invasion.