Effects of HSP47 siRNA on biological behaviors of HTCF cells cultured in vitro and the expression level of TGF-β1

Abstract

AIM: To explore the effects of heat shock protein 47(HSP47)siRNA on biological behaviors of human Tenon capsule fibroblasts(HTCF)cells cultured in vitro and the expression level of transforming growth factor-β1(TGF-β1).METHODS: HTCF were cultured in vitro and divided into blank control group, empty vector group and transfection group. In transfection group, interfering siRNA sequences were designed and synthesized based on the HSP47 gene sequences, vectors were constructed and introduced into HTCF. The empty vector group was introduced with empty vectors. The expressions of HSP47 mRNA and protein in cells were detected by RT-PCR and Western blot. The proliferation, apoptosis, invasion and migration of cells were detected by clone formation assay, flow cytometry, Transwell method and scratch test. The expressions of proliferation, apoptosis, invasion and migration proteins, and TGF-β1 were detected by Western blot.RESULTS: Compared with empty vector group, expression of HSP47 mRNA and protein, clone formation rate, cell healing rate, number of invasive cells, relative expression levels of Ki67, N-cadherin and TGF-β1 were significantly decreased in transfection group(P0.05).CONCLUSION: HSP47 siRNA can reduce proliferation, invasion and migration abilities of HTCF cells by inhibiting the expression of TGF-β1 protein, without significant effects on the apoptosis of HTCF cells.

Keywords

• heat shock protein 47
• human tenon capsule fibroblast cell
• transforming growth factor-β1