摘要
为探讨胃癌差异表达蛋白MAWD和MAWBP对胃癌细胞EMT的影响。采用pc DNA3.1构建MAWD真核表达载体,在胃癌细胞SGC7901中单独和共同过量表达MAWD与MAWBP,采用免疫荧光检测MAWD和MAWBP在细胞中的定位,采用western blot检测EMT相关标志蛋白E-cadherin、N-cadherin和Snail在细胞中的表达水平。结果显示,成功构建过量表达载体并稳定转染细胞,转染MAWD和MAWBP后,SGC7901细胞形态呈现类似上皮细胞的鹅卵石状,单独转染Vector的细胞则呈现纤维状,MAWD和MAWBP在胞核和胞浆中均有分布,E-cadherin在MAWD和MAWBP共同过量表达细胞中表达明显增强,N-cadherin和Snail则表达减弱。由此可知,MAWD和MAWBP协同抑制胃癌细胞中的EMT。讨论:研究发现MAWD通过Smad7形成复合体加强对TGF-beta通路的抑制,而TGF-beta是诱导EMT的重要细胞因子。MAWBP能与MAWD相互结合,二者共同过量表达时,通过协同作用影响TGF-beta相关的EMT标志性蛋白表达。
To explore the effect of differential expression of Mitogen-Activated Protein Kinase Activator with WD40 repeats（MAWD） and MAWD binding protein（MAWBP） on epithelial-mesenchymal transition（EMT） of gastric cancer（GC） cells. The eukaryotic expression vector was constructed by pc DNA3.1,and transfected MAWD and MAWBP to GC cells SGC7901. The morphologic change of the cell and the location of MAWD and MAWBP was observed by immunofluorescence. The expression level of EMT related protein E-cadherin, N-cadherin and Snail was detected by western blot. The eukaryotic expression vector was successfully constructed and transfected to the SGC7901 cells. The cellular morphology was changed from fibrous shape to the pebble shape after transfected with MAWD and MAWBP. MAWD and MAWBP was located in the cytoplasm and the cell nucleus. E-cadherin was obviously over expressed in MAWD and MAWBP co-expressed cells, while N-cadherin and Snail was reduced. MAWD and MAWBP could synergistically suppressed the EMT in GC cells. Discussion： MAWD inhibit TGF-beta signal through Smad7. TGF-beta is an important cytokine for EMT. MAWBP can form the complex with MAWD.Co-overexpression of MAWD and MAWBP can synergistically affect the expression of EMT protein who is related with TGF-beta signaling pathway.
出处
《石河子大学学报（自然科学版）》
CAS
2015年第1期92-96,共5页
Journal of Shihezi University(Natural Science)
基金
国家自然科学基金项目(30901717)
