HNF1α inhibition triggers epithelial-mesenchymal transition in human liver cancer cell lines

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作者：

V Danijela，R Sandra，P Laura，BS Paulette，ZR Jessica

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摘要：

Background Hepatocyte Nuclear Factor 1?? (HNF1??) is an atypical homeodomain-containing transcription factor that transactivates liver-specific genes including albumin, ??-1-antitrypsin and ??- and ??-fibrinogen. Biallelic inactivating mutations of HNF1A have been frequently identified in hepatocellular adenomas (HCA), rare benign liver tumors usually developed in women under oral contraceptives, and in rare cases of hepatocellular carcinomas developed in non-cirrhotic liver. HNF1??-mutated HCA (H-HCA) are characterized by a marked steatosis and show activation of glycolysis, lipogenesis, translational machinery and mTOR pathway. We studied the consequences of HNF1?? silencing in hepatic cell lines, HepG2 and Hep3B and we reproduced most of the deregulations identified in H-HCA. Methods We transfected hepatoma cell lines HepG2 and Hep3B with siRNA targeting HNF1?? and obtained a strong inhibition of HNF1?? expression. We then looked at the phenotypic changes by microscopy and studied changes in gene expression using qRT-PCR and Western Blot. Results Hepatocytes transfected with HNF1?? siRNA underwent severe phenotypic changes with loss of cell-cell contacts and development of migration structures. In HNF1??-inhibited cells, hepatocyte and epithelial markers were diminished and mesenchymal markers were over-expressed. This epithelial-mesenchymal transition (EMT) was related to the up regulation of several EMT transcription factors, in particular SNAIL and SLUG. We also found an overexpression of TGF??1, an EMT initiator, in both cells transfected with HNF1?? siRNA and H-HCA. Moreover, TGF??1 expression is strongly correlated to HNF1?? expression in cell models, suggesting regulation of TGF??1 expression by HNF1??. Conclusion Our results suggest that HNF1?? is not only important for hepatocyte differentiation, but has also a role in the maintenance of epithelial phenotype in hepatocytes.

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关键词：

siRNA hepatocellular adenoma tumor suppressor gene benign tumor Hepatocyte Nuclear Factor 1 EMT TGF??1 Oncology Internal Medicine Medicine(General