小鼠神经母细胞瘤细胞中神经型钙黏蛋白超表达对突起形成及连环蛋白表达的影响

朱少义 管丽红* 杨慈清 张必超 李秋玲 林俊堂* 

doi:10.16098/j.issn.0529-1356.2018.05.006

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解剖学报 ›› 2018, Vol. 49 ›› Issue (5) : 598-604. DOI: 10.16098/j.issn.0529-1356.2018.05.006

细胞和分子生物学

小鼠神经母细胞瘤细胞中神经型钙黏蛋白超表达对突起形成及连环蛋白表达的影响

朱少义^1,2 管丽红^1,2* 杨慈清^1,2 张必超^1,2 李秋玲^1,2 林俊堂^1,2,3*

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Effect of N-cadherin overexpression on protrusion formation and catenins expression in mouse neuroblastoma N2a cells

ZHU Shao-yi ^1,2 GUAN Li-hong ^1,2* YANG Ci-qing ^1,2 ZHANG Bi-chao ^1,2 LI Qiu-ling ^1,2 LIN Jun-tang ^1,2,3*

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摘要

目的探讨神经型钙黏蛋白（N-cadherin）调控连环蛋白（catenin）对小鼠神经母细胞瘤细胞（N2a）形态的影响。方法构建鼠源的N-cadherin融合蛋白超表达载体，利用脂质体转染的方法将空载体pCAG-MCS-EGFP和超表达载体pCAG-N-Cad-EGFP分别转染至N2a细胞，作为对照组和实验组；细胞划痕和黏附实验检测N-cadherin超表达对N2a细胞迁移和黏附的影响，免疫荧光检测N-cadherin超表达对N2a细胞形态的影响，并结合Western blotting检测连环蛋白家族的表达情况。结果与对照组相比，N-cadherin超表达促进细胞突起的形成，使得N2a细胞突起增多和伸长；N-cadherin超表达增强了黏附性而抑制了细胞的迁移能力；N-cadherin超表达使得细胞内连环蛋白家族中α-catenin、β-catenin和p120蛋白表达均显著上升（P<0.05），而γ-catenin表达下降但差异不显著（P>0.05）；另外，N-cadherin超表达的细胞有明显的聚集现象。结论 N-cadherin超表达促进N2a细胞突起形成，增强细胞的黏附性从而抑制细胞的迁移能力，调控连环蛋白家族成员的表达并促使细胞聚集。

Abstract

Objective To analyse the effects of N-cadherin overexpression on cell morphology and catenins expression by using mouse neuroblastoma N2a cells. Methods The mouse N-cadherin gene was cloned to pCAG-MCS-EGFP plasmid and transfected into N2a cells by Lipofectamine 3000. The empty vector pCAG-MCS-EGFP and the overexpression vector pCAG-N-Cad-EGFP transfected group were used as control group and experimental group, respectively. Cell scratche and cell adhesion experiments were used to detect the effects of N-cadherin overexpression on the migration and adhesion of N2a cells. Immunofluorescence was used to detect the effect of N-cadherin overexpression on the morphology of N2a cells. The expression of catenin family was detected by Western blotting. Results Compared with the control group, overexpressing N-cadherin promoted the protrusion formation of N2a cells, which led to the increase and elongation of protrusion in N2a cells. N-cadherin overexpression enhancea cell adhesion and inhibited migration. N-cadherin overexpression resulted in the expression level of α-catenin, β-catenin and p120 protein significantly increased (P<0.05), while γ-catenin expression decreased but the difference was not significant (P>0.05). In addition, the cell aggregation phenomenon was observed when N-cadherin overexpression. Conclusion N-cadherin overexpression promotes protrusion formation and adhesion ability in N2a cells, but inhibits cell migration, regulates the expression of the family of catenin and promotes cell aggregation.

导出引用

朱少义管丽红杨慈清张必超李秋玲林俊堂. 小鼠神经母细胞瘤细胞中神经型钙黏蛋白超表达对突起形成及连环蛋白表达的影响[J]. 解剖学报. 2018, 49(5): 598-604 https://doi.org/10.16098/j.issn.0529-1356.2018.05.006

ZHU Shao-yi GUAN Li-hong YANG Ci-qing ZHANG Bi-chao LI Qiu-ling LIN Jun-tang. Effect of N-cadherin overexpression on protrusion formation and catenins expression in mouse neuroblastoma N2a cells[J]. Acta Anatomica Sinica. 2018, 49(5): 598-604 https://doi.org/10.16098/j.issn.0529-1356.2018.05.006

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