| Summary: | 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.
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