SHP-2激活突变促进小鼠MEFs细胞粘附迁移及增殖能力

目的探讨SHP-2D61G/＋和SHP-2D61G/D61G激活突变对小鼠胚胎成纤维细胞（MEFs）粘附迁移及增殖能力的影响,并研究其发生的机制。方法雌雄小鼠合笼交配建立SHP-2D61G/＋、SHP-2D61G/D61G激活突变的小鼠MEFs细胞,并以SV40T抗原进行永生化;细胞粘附实验检测SHP-2D61G/＋、SHP-2D61G/D61G激活突变对MEFs细胞粘附能力的影响;Transwell体外迁移实验检测SHP-2D61G/＋、SHP-2D61G/D61G激活突变对MEFs细胞的迁移能力的影响;MTT法检测SHP-2D61G/＋、SHP-2D61G/D61G激活突变对MEFs细胞增殖能力的影响;Western Blot法检测p-ERK的表达水平。结果 （1）与对照组相比,SHP-2D61G/＋、SHP-2D61G/D61G激活突变组小鼠MEFs细胞粘附的细胞数明显增多,差异具有统计学意义;（2）与对照组相比SHP-2D61G/＋、SHP-2D61G/D61G激活突变组MEFs细胞迁移的细胞数增加,差异具有统计学意义;（3）MTT结果显示,SHP-2D61G/＋、SHP-2D61G/D61G激活突变的小鼠MEFs细胞增殖能力较对照组强,差异具有统计学意义;（4）Western Blot结果显示与对照组相比,无论是刚刚贴壁还是贴壁后30 min和60 min SHP-2D61G/＋、SHP-2D61G/D61G激活突变组其p-ERK的表达水平都增加。结论 SHP-2D61G/＋、SHP-2D61G/D61G激活突变促进小鼠MEFs细胞粘附迁移及增殖能力,其发生机制主要与p-ERK的表达水平增加有关。

SHP-2 gain-of-function mutations promote adhesion,migration and proliferation abilities of mouse embryonic fibroblasts

Objective To explore the effects of SHP-2D61G/＋ and SHP-2D61G/D61G gain-of-function mutations on cell adhesion,migration and proliferation abilities of mouse embryonic fibroblasts（ MEFs）,and to study their mechanism of action. Methods Male and female mice were mated to establish SHP-2D61G /＋ and SHP-2D61G / D61G gain-of-function mutation MEFs and then immortalized by SV40T antigen. Cell adhesion assay was used to detect the effects of SHP-2D61G / ＋ and SHP-2D61G /D61G gain-of-function mutations on adhesion ability of MEFs. Transwell migration assay in vitro was applied to detect the effects of SHP-2D61G /＋ and SHP-2D61G /D61G gain-of-function mutations on the migration ability of MEFs. MTT assay was used to assess the effects of SHP-2D61G / ＋ and SHP-2D61G / D61G gain-of-function mutations on the proliferation ability of MEFs. Western Blot assay was conducted to detect p-ERK expression levels. Results（1） Compared with the control group,SHP-2D61G /＋ and SHP-2D61G / D61G gain-of-function mutations of MEFs significantly increased the number of adherent cells（ P 0. 05,P 0. 01）;（ 2） Compared with the control group,SHP-2D61G /＋ and SHP-2D61G / D61G gain-of-function mutations of MEFs significantly increased the number of migration cells（ P 0. 05,P 0. 01）;（ 3） MTT assay showed that SHP-2D61G /＋ and SHP-2D61G / D61G gain-of-function mutations of MEFs significantly enhanced the proliferation ability than that in the control group（ P 0. 05,P 0. 01）;（4） Western Blot showed that compared with the control group,whether the MEFs were just adherent or 30 min and 60 min after adhesion,p-ERK expression level was significantly increased in the MEFs with SHP-2D61G /＋ and SHP-2D61G / D61G gain-of-function mutations. Conclusions SHP-2D61G /＋ and SHP-2D61G / D61G gain-of-function mutations promote the cell adhesion and migration and proliferation abilities of mouse embryonic fibroblasts. Its mechanism is mainly related with the increase of p-ERK expression level.