Effect ofHepatocyte nuclear factor 4 on stem cell characteristics and epithelial mesenchymal transition in hepatocellular carcinoma cells

Abstract

Objective To investigate the effect of hepatocyte nuclear factor 4 (HNF4) on the characteristics of stem cells and epithelial mesenchymal transition in hepatocellular carcinoma cell line SMMC7721. Methods Thehoechst33342 staining and CD133 staining, followed by flow cytometry sorting were performed in SMMC7721 cells. The expression levels of HNF4 were detected by Western blotting in the side group cells and CD133+ cells. Western blotting was used to detect the protein changes, which related tothe epithelial mesenchymal transition, after HNF4 was knockdown by specific siRNA transfection. Finally, the flow cytometry and in vitro cell sphere formation assay were conducted to value the effect of HNF4 silencing on stem cell properties. Results The side group cells, resembled by Hoechst33342 negative, accounted (8.77±0.59)% of the proportion of total cells, while the proportion of CD133+ cells was (4.19±0.26)% of the total cells, the expression levels of HNF4 were significantly lower in the side population cells and CD133+ cells, and the HNF4 silencing could promote the expression of epithelial cell marker protein, with the decline of mesenchymal ones. Knockdown of HNF4 could also elevate the proportion of side group cells (NC vs. si-HNF4=7.73±0.57 vs. 22.38±2.56) and CD133+ cells (NC vs. si-HNF4=4.67±0.56 vs. 17.31±1.77), and also promotedthe activity of SMMC7721 cell sphere formation (NC vs. si-HNF4=1.67±0.62 vs. 5.33±1.52). Conclusion HNF4 plays an important role in maintaining the stem cell properties of HCC cells. Silencing HNF4 could promote the epithelial mesenchymal transition, and enhance the stem cell characteristics of hepatocellular carcinoma cells. Key words: Hepatocyte nuclear factor 4; Stem cells; Epithelial mesenchymal transition