[1]黄玉婷,董金玲,冯力元,等.内源性过表达Nestin对NIH3T3细胞增殖、迁移及MAPK信号传导通路的影响[J].第三军医大学学报,2020,42(02):125-132.
 HUANG Yuting,DONG Jinling,FENG Liyuan,et al.Effects of endogenous nestin overexpression on proliferation, migration and MAPK signal transduction pathway in NIH3T3 cells[J].J Third Mil Med Univ,2020,42(02):125-132.
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内源性过表达Nestin对NIH3T3细胞增殖、迁移及MAPK信号传导通路的影响(/HTML )
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《第三军医大学学报》[ISSN:1000-5404/CN:51-1095/R]

卷:
42卷
期数:
2020年第02期
页码:
125-132
栏目:
基础医学
出版日期:
2020-01-30

文章信息/Info

Title:
Effects of endogenous nestin overexpression on proliferation, migration and MAPK signal transduction pathway in NIH3T3 cells
作者:
黄玉婷董金玲冯力元李鹏
400038 重庆,陆军军医大学(第三军医大学)药学与检验医学系生药学与中药学教研室
Author(s):
HUANG Yuting DONG Jinling FENG Liyuan LI Peng

Department of Pharmacognosy and Traditional Chinese Medicine, Faculty of Pharmacy and Laboratory Medicine, Army Medical University (Third Military Medical University), Chongqing, 400038, China
 

关键词:
NestinNIH3T3CRISPR/SAM增殖迁移MAPK
Keywords:
nestin NIH3T3 CRISPR/SAM proliferation migration mitogen-activated protein kinase
分类号:
R329-33;R329.28;R341
文献标志码:
A
摘要:

目的通过在成纤维细胞NIH3T3中内源性过表达巢蛋白(Nestin),研究其对细胞增殖、迁移及转化生长因子(TGF-β1)作用下细胞胶原蛋白表达与MAPK相关通路的影响。方法通过CRISPR/SAM技术构建内源性过表达Nestin 的细胞株,用MTS实验、Western blot检测细胞增殖能力,细胞划痕实验检测细胞迁移能力,经qPCR、Western blot与免疫荧光染色明确TGF-β1对其胶原蛋白与MAPK通路的影响。结果CRISPR/SAM技术能使内源性Nestin表达显著提高(P<0.05),而内源性过表达Nestin对NIH3T3细胞的生长、迁移有明显促进作用(P<0.05),对TGF-β1诱导的胶原蛋白表达及MAPK的激活也有明显的抑制效应(P<0.01)。结论用CRISPR/SAM技术内源性过表达Nestin 可以明显促进NIH3T3细胞的生长和迁移,但却抑制TGF-β1诱导下胶原蛋白的表达及MAPK的激活。

Abstract:

ObjectiveTo 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. MethodsCRISPR/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. ResultsCRISPR/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). ConclusionEndogenous 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.

参考文献/References:

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更新日期/Last Update: 2020-01-29