摘要
目的:实现鸡源N-Cadherin在293T细胞中的过表达及其抑制表达,进一步分析N-Cadherin异常表达对293T细胞内源性β-catenin和α-catenin的表达及细胞形态结构的影响。方法:采用脂质体转染法,设对照、过表达和抑制表达3组,将pCAGGS-GFP、pCAGGS-N-Cadherin和pCAGGS-N-Cadherin/pGPU6-GFP-neo-N-Cadherin-shRNA质粒分别转染到293T细胞中,转染后48小时观察拍照,荧光免疫组化检测N-Cadherin和相关蛋白表达情况。结果:N-Cadherin过表达后会改变293T细胞的形态,其细胞突起明显增加,而对N-Cadherin干扰后,突起又会消失,并且外源性N-Cadherin的过表达会使得293T细胞内源性β-catenin和α-catenin的表达升高,抑制其表达β-catenin和α-catenin表达也随之减弱,并表现出β-catenin在核内的聚集。结论:异源性N-Cadherin的表达明显改变293T细胞的形态,并且内源性β-catenin和α-catenin的表达与N-Cadherin的表达呈正相关。
Objective：To achieve the chicken source N-Cadherin over-expression and suppression of expression in 293T cells,further analysis N-cadherin abnormal expression cells endogenous beta-catenin and alpha-catenin expression and cell morphology changes.Methods：Liposome transfection method was used in the experiment,pCAGGS-GFP,pCAGGS-N-Cadherin and pCAGGS-N-cadherin/pGPU6-GFP-neo-N-Cadherin-shRNA plasmid transfected into 293T cells,after transfection 48 h take photos and immunofluorescence detection of N-cadherin and protein expression.Results：The N-Cadherin over-expression change 293T cell morphology,the projection among the cells increased significantly,while the N-Cadherin interference the projection will disappear.The exogenous N-Cadherin overexpression would increase endogenous beta-catenin and alpha-catenin expression in 293T cells,inhibition of the N-Cadherin expression would lead to beta-catenin and alpha-catenin expression weakens,and exhibit aggregation of beta-catenin in the nucleus.Conclusion：N-Cadherin expression will change the morphology of 293T cells significantly,and the endogenous beta-catenin and alpha-catenin expression has positive correlate with the N-cadherin expression.
出处
《中国免疫学杂志》
CAS
CSCD
北大核心
2013年第9期914-917,共4页
Chinese Journal of Immunology
基金
2010年国家自然科学基金(No.31000475)
2013年河南省教育厅科学技术研究重点项目(No.13A320866)
2011年新乡医学院重点研究领域招标课题资助项目(No.ZD2011-26)
