Enhancer of zeste homolog 2 (EZH2) gene inhibition via 3-Deazaneplanocin A (DZNep) in human liver cells and it is relation with fibrosis

Abstract

Abstract Objectives Hepatic fibrosis is a complex and dynamic process, such as “wound healing”. The effect of 3-deazaneplanocin A (DZNep) via enhancer of zeste homolog 2 (EZH2) inhibition on transforming growth factor, matrix metalloproteinases 2-9 (MMP2, MMP9) and matrix metalloproteinases inhibitor 3 (TIMP3), alpha-smooth muscle actin (α-SMA), collagen 1A1(COL1A1), and collagen 3A1 (COL3A1) genes/proteins in human hepatic stellate cell line were examined. Inhibition effect of EZH2 on colony formation and migration were investigated. Methods 5 μM DZNep was treated to LX2 cells for 3 days. Real time PCR and Western blot method were used for analyses. Results DZNep was shown to reduce colony formation and migration. It increased the α-SMA only in gene expression level but decreased at the protein levels. It down-regulated both gene expression and protein levels of EZH2, while up-regulated gene expression and protein levels of TGF-β. DZNep induced gene expression and protein levels of MMP2 and TIMP3. However, DZNep induced COL1A1 and COL3A1 gene expression levels but reduced protein levels only in COL3A1 but not COL1A1. Conclusion DZNep has a positive anti-fibrotic effect by reducing α-SMA and COL3A1 protein levels in the LX2 cell line, however, we have found that DZNep may also has fibrotic effect because it increases TGF-β both expression and protein levels.