miR-26b通过调节Notch1信号通路参与原发性肝细胞肝癌侵袭的机制研究

目的:探讨miR-26b参与原发性肝细胞肝癌(HCC)侵袭的机制。方法:在细胞培养液中培养人肝细胞系HL-7702和HCC细胞各系Hepb-3、HuH-7、MHCC97-L、MHCC97-H。实时荧光定量PCR法(qRT-PCR)检测miR-26b的表达水平;用miR-26b mimics、miR-26b inhibitors和Notch1-siRNA分别转染HCC细胞;MTT实验检测转染后HCC细胞的活力;采用Western blot检测Notch1受体蛋白表达水平的变化;Transwell小室测定不同处理后的HCC细胞的侵袭能力。结果:人正常肝细胞系HL-7702和HCC细胞系Hepb-3、HuH-7、MHCC97-L、MHCC97-H中的miR-26b相对表达含量随其侵袭和迁移能力的升高而依次下降;抑制miR-26b的表达,Notch1受体蛋白表达明显增高,而此时HCC细胞的侵袭性显著增强;相反,上调miR-26b的表达,Notch1受体蛋白表达明显降低,而HCC细胞侵袭性显著下降;miR-26b可能通过调控Notch1信号通路调节HCC细胞侵袭性。结论:miR-26b通过负调控Notch1信号通路抑制HCC细胞侵袭能力,为HCC侵袭的机制奠定了理论基础,miR-26b可能成为HCC治疗的新靶点。

miR-26b is involved in cell invasion in hepatocellular carcinoma by regulating the Notch1 signal pathway

Objective:To investigate the involvement of miR-26b in the invasion and metastasis of primary hepatocellular carcinoma (HCC).Methods:HL-7702 human hepatocytes and hepatocellular carcinoma cells were cultured in cell culture medium.Each line of Hepb-3,HuH-7,MHCC97-L,MHCC97-H.MTT assay was used to detect the viability of HCC cells.The expression level of miR-26b was determined by qRT-PCR.HCC cell lines were transfected with miR-26b inhibitor,miR-26b mimic and Notch1-siRNA.The protein expression level was detected by Western blot.Transwell assay was used to determine the invasion ability of cell lines after different treatments.Results:The relative expression levels of miR-26b in HL-7702 human normal liver cell line and Hepb-3,HuH-7,MHCC97-L and MHCC97-H cell lines decreased with the increase of invasion and migration ability.By inhibiting the expression of miR-26b,the protein expression of Notch1 receptor was significantly increased,while the invasiveness of HCC cells was significantly enhanced.On the contrary,when the expression of miR-26b was upregulated,the protein expression of Notch1 receptor was significantly decreased,while the invasiveness of HCC cells was significantly decreased.miR-26b may regulate the invasiveness of HCC cells by regulating the Notch1 signaling pathway.Conclusion:miR-26b inhibits liver cancer cell metastasis through negative regulation of Notch1 signaling pathway,laying a theoretical foundation for the mechanism of liver cancer metastasis.miR-26b may become a new target for the treatment of primary hepatocellular carcinoma.