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

Abstract : Background
Hepatocyte Nuclear Factor 1α (HNF1α) is an atypical homeodomain-containing transcription factor that transactivates liver-specific genes including albumin, α-1-antitrypsin and α- and β-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. HNF1α-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 HNF1α 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 HNF1α and obtained a strong inhibition of HNF1α 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 HNF1α siRNA underwent severe phenotypic changes with loss of cell-cell contacts and development of migration structures. In HNF1α-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 TGFβ1, an EMT initiator, in both cells transfected with HNF1α siRNA and H-HCA. Moreover, TGFβ1 expression is strongly correlated to HNF1α expression in cell models, suggesting regulation of TGFβ1 expression by HNF1α.
Conclusion
Our results suggest that HNF1α is not only important for hepatocyte differentiation, but has also a role in the maintenance of epithelial phenotype in hepatocytes.
Type de document :
Article dans une revue
BMC Cancer, BioMed Central, 2011, 11 (1), pp.427. 〈10.1186/1471-2407-11-427〉
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http://www.hal.inserm.fr/inserm-00636820
Contributeur : Ed. Bmc <>
Soumis le : vendredi 28 octobre 2011 - 13:10:49
Dernière modification le : jeudi 11 janvier 2018 - 06:15:14
Document(s) archivé(s) le : jeudi 15 novembre 2012 - 10:51:52

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Laura Pelletier, Sandra Rebouissou, Danijela Vignjevic, Paulette Bioulac-Sage, Jessica Zucman-Rossi. HNF1α inhibition triggers epithelial-mesenchymal transition in human liver cancer cell lines. BMC Cancer, BioMed Central, 2011, 11 (1), pp.427. 〈10.1186/1471-2407-11-427〉. 〈inserm-00636820〉

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