遗传印记基因PEG10沉默对人肝癌细胞Wnt/β-catenin信号转导通路的影响

The effect of genetic imprintined gene PEG10 silencing on Wnt/β-catenin signal transduction pathway in human liver cancer cell

目的观察靶向封闭遗传印记基因PEG10对人肝癌细胞(HepG2)Wnt/β-catenin信号转导通路的影响,探讨PEG10促进肝癌形成的可能机制。方法设计针对PEG10基因的shRNA,体外转录制备shRNA后利用脂质体转染技术转染肝癌细胞株HepG2,RT-PCR检测PEG10、β-catenin(CTNNB1)、WNT1基因mRNA水平,应用免疫组化技术检测其蛋白表达水平。结果转染PEG10-shRNA后HepG2细胞PEG10 mRNA水平下降65.6%,其蛋白表达水平减少60.9%;同时,β-catenin(CTNNB1)、WNT1基因mRNA水平随之分别下降52.5%、96.1%,蛋白表达水平分别减少94.3%、63.2%。结论靶向封闭PEG10可下调肝癌细胞Wnt/β-catenin信号通路关键因子β-catenin(CTNNB1)、WNT1表达水平。PEG10对肝癌的促进作用可能部分与其影响Wnt/β-catenin通路有关。

Objective To observe the blockage effect of genetic imprinted gene PEG10 on Wnt/β-catenin signal transduction pathway in liver cancer cell line HepG2,and to explore the possible mechanism of the carcinogenesis of PEG10 in liver cancer.Methods PEG10-shRNAs were designed and synthesized through transcription in vitro.PEG10-shRNAs were transfected into HepG2 cells through liposome.The mRNA levels of PEG10,β-catenin（CTNNB1） and WNT1 gene were detected by RT-PCR,and their protein levels were detected by immunohistochemical staining.Results After transfected by PEG10-shRNA,the PEG10 mRNA level in HepG2 cells reduced by 65.6%.At the same time,the mRNA levels of β-catenin（CTNNB1） and WNT1 reduced by 52.5%,96.1%,respectively.Immunohistochemical staining showed that after transfected by PEG10-shRNA,the PEG10 protein level in HepG2 cells reduced by 60.9%.At the same time,the protein levels of β-catenin（CTNNB1） and WNT1 reduced by 94.3%,63.2%,respectively.Conclusion The blockage of PEG10 in HepG2 cells downregulates the expression of β-catenin（CTNNB1） and WNT1,which are the key factors of Wnt/β-catenin signaling pathway.The carcinogenesis of PEG10 in liver cancer may partly owe to its effect on Wnt/β-catenin pathway.