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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
Effects of endogenous nestin overexpression on proliferation, migration and MAPK signal transduction pathway in NIH3T3 cells
- 第三军医大学学报 2020年42卷第2期 页码：125-132
- 作者机构：
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
- 作者简介：
[李鹏, E-mail: pengli@tmmu.edu.cn LI Peng, E-mail: pengli@tmmu.edu.cn
- 基金信息：国家自然科学基金面上项目(81472596)
DOI：10.16016/j.1000-5404.201908194
中图分类号： R329-33;R329.28;R341收稿：2019-08-27，
修回：2019-11-20，
纸质出版：2020-01-30
- 稿件说明：
移动端阅览
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(2):125-132.
Yuting HUANG, Jinling DONG, Liyuan FENG, et al. Effects of endogenous nestin overexpression on proliferation, migration and MAPK signal transduction pathway in NIH3T3 cells[J]. Journal of Third Military Medical University, 2020, 42(2): 125-132.
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(2):125-132. DOI： 10.16016/j.1000-5404.201908194.
Yuting HUANG, Jinling DONG, Liyuan FENG, et al. Effects of endogenous nestin overexpression on proliferation, migration and MAPK signal transduction pathway in NIH3T3 cells[J]. Journal of Third Military Medical University, 2020, 42(2): 125-132. DOI： 10.16016/j.1000-5404.201908194.
摘要
目的
2
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
2
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
2
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
2
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
Objective
2
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
2
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
2
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
2
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.
关键词
Keywords
references
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