Effects of endogenous nestin overexpression on proliferation, migration and MAPK signal transduction pathway in NIH3T3 cells

Abstract

Objective To observe the effects of endogenous nestin overexpression on the proliferation, migration and transforming growth factor β (TGF-β1)-induced expression of collagen and MAPK-related pathway in NIH3T3 fibroblasts. Methods CRISPR/SAM technique was used to construct a nestin-overexpressing cell line in NIH3T3 fibroblasts. The effects of nestin overexpression on cell proliferation and migration were evaluated using MTS assay, immunoblotting and scratch-wound assay. Quantitative PCR, immunoblotting and immunofluorescence staining were used to determine the effect of nestin overexpression on TGF-β-induced expression of collagen and activation of MAPK pathway in the fibroblasts. Results CRISPR/SAM significantly increased the expression of endogenous nestin in NIH3T3 fibroblasts (P < 0.05), and nestin overexpression significantly promoted the growth and migration of NIH3T3 cells (P < 0.05). Nestin overexpression obviously inhibited the expression of collagen and the activation of MAPK in TGF-β1-induced cells (P < 0.01). Conclusion Endogenous overexpression of nestin achieved by CRISPR/SAM technique significantly promotes the growth and migration of NIH3T3 cells and inhibits TGF-β1-induced expression of collagen and activation of MAPK.

Keywords

• nestin
• nih3t3
• crispr/sam
• proliferation
• migration
• mitogen-activated protein kinase