miR-21通过调节PTEN及Wnt信号通路介导Huh7细胞对顺铂的耐药性

Regulation of microRNA-21 for cisplatin resistance in Huh7 cells by PTEN and Wnt signaling pathways

目的探讨microRNAs(miRNAs)介导肝细胞癌(HCC)对顺铂的耐药机制。方法采用miR-21 mimics及miR-21抑制剂来调节Huh7细胞miR-21表达量。人DKK-1与bpv(phen)分别是Wnt信号通路抑制剂及PTEN抑制剂。相关mRNA及蛋白水平采用RT-PCR及蛋白质免疫检测。结果 miR-21过表达将减弱顺铂(5和10μg/mL)对Huh7细胞增殖的抑制作用。当Huh7细胞经miR-21 mimics处理后,Wnt4的mRNA及蛋白质表达均显著上升,且miR-21过表达可逆转DKK-1对Wnt4和细胞增殖的抑制作用。另外,miR-21过表达还上调了PTEN的mRNA及蛋白表达,并可提升经bpv(100和200nmol/L)抑制的Huh7细胞增殖速率。结论 miR-21在Huh7细胞对顺铂的耐药中发挥作用,且通过活化Wnt信号通路及PTEN调节Huh7细胞增殖。

Objective To investigate the mechanism of microRNAs in mediating cisplatin resistance in hepatocellular carcinoma（HCC）.Methods MicroRNA-21（miR-21）level was determined in HCC patients after cisplatin chemotherapy.miR-21 mimics and miR-21 inhibitor were used to upregulate or downregulate miR-21 level in Huh-7 cells.Human dickkopf-1（DKK-1）and bpV（phen）were used as Wnt inhibitor and phosphate and tension homology deleted on chromsome ten（PTEN）inhibitor,respectively.Relative mRNA level and protein expression were detected by real-time polymerase chain reaction（RT-PCR）and western blot,respectively.Results The results showed that overexpression of miR-21 decreased cisplatin（5 and10μg/mL）-induced inhibition of cell proliferation in Huh7 cells.Both mRNA level and protein expression of Wnt4 were elevated remarkably when Huh7 cells were treated with miR-21 mimics.Moreover,inhibition of Wnt4 and cell proliferation by DKK-1 was reversed using overexpression of miR-21.In addition,miR-21 mimics increased both mRNA level and protein expression of PTEN,and overexpression of miR-21 increased cell proliferation rate which was inhibited using bpV（phen,100 and200 nmol/L）in Huh7 cells.Conclusion miR-21 was involved in regulating resistance to cisplatin in Huh7 cells by activating Wnt and PTEN signaling pathways.