LncRNA-NEF transcriptionally activates FOXA2 and inhibits epithelial–mesenchymal transition of hepatocellular carcinoma cells by inhibiting β-catenin signaling pathway

Abstract

Abstract: Objective To explore the molecular mechanism of long-chain non-coding RNA-neighboring enhancer of FOXA2 (lncRNA-NEF) on the epithelial-mesenchymal transition (EMT) process of hepatocellular carcinoma (HCC), a key step in liver cancer metastasis. Methods HCC line Hep3B was treated with transforming growth factor-β (TGF-β) to induce EMT. A pcDNA3.1-FOXA2 overexpression vector was constructed and transduced into another HCC line HepG2 to overexpress FOXA2 and induce mesenchymal-epithelial transition (MET). The EMT-related markers including E-cadherin, Vimentin and Snail were detected and cell invasion assay was performed in these lines with and without relative treatment. The expression of lncRNA-NEF was determined in 10 paired operated liver cancer tissues and adjacent normal tissues, as well as in the HCC lines. Subsequently, the protein expression of factors related to β-catenin signaling pathway before and after EMT or MET transformation in the HCC lines were detected. Results The expression level of lncRNA-NEF in liver cancer tissues and HCC lines was lower than that in normal controls (P<0.05). Compared with the control cells, the cell invasion ability of TGF-β treated HCC cells significantly enhanced (P<0.05), but the cell invasion ability of PCDNA3.1-FOXA2 transduced cells significantly reduced (P<0.05). The expression level of phosphorylated β-catenin (p-β-catenin) in Hep3B cells treated with TGF-β increased significantly when compared with the control cells. On the contrary, the expression level of phosphorylated β-catenin (P-β-catenin) in HepG2 cells transfected with PCDNA3.1-FoxA2 significantly decreased. The level of total β-catenin protein remained unchanged in both HCC lines with and without relative treatment. LncRNA-NEF activated FOXA2 in a cis-acting manner, and inhibited the EMT process of liver cancer cells by inhibiting the β-catenin signaling pathway. Conclusion LncRNA-NEF activates FOXA2 transcription and inhibits the EMT process of liver cancer cells by inhibiting the β-catenin signaling pathway.

Key words: LncRNA-NEF, FOXA2, EMT, MET, β-catenin signaling pathway

CHE Jun, YANG Liu-qing, SUN Bin, JIA Ze-bo. LncRNA-NEF transcriptionally activates FOXA2 and inhibits epithelial–mesenchymal transition of hepatocellular carcinoma cells by inhibiting β-catenin signaling pathway[J]. Chinese Hepatolgy, 2021, 26(10): 1137-1141.

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