摘要
目的探索Nesprin-1在连接细胞骨架与细胞核中的作用及对细胞迁移的影响。方法通过基因敲除的方法建立Nesprin-1缺失MEFs细胞,并通过PCR及Western blot进行验证;细胞计数实验检测敲除Nesprin-1对细胞生长的影响;细胞免疫荧光分析细胞骨架蛋白结构及微管组织中心(MTOC)定位;划痕实验用来检测细胞迁移情况。结果 Nesprin-1缺失MEFs细胞株建立成功,其生长曲线与野生型细胞一致,但细胞骨架蛋白F-actin丝状结构消失,微管蛋白表达减少,MTOC与细胞核分离,且细胞迁移能力降低。结论Nesprin-1通过连接核骨架和细胞骨架调控细胞骨架硬度及MTOC极化,从而影响细胞迁移。
Objective This study aimed to investigate the role of Nesprin-1 in connection of nucleus and cytoskeleton and controlling cell migration. Methods We established Nesprin-1 deficient MEF cells by knockout mouse model and confirmed by PCR and Western blot. The cell viability was detected by cell counting and growth curve plotting. The position of MTOC and organization of cytoskeleton was performed by immunocytofluorescence. And the woundhealing experiment was product to observe cell migration. Results We had successfully established Nesprin-1 null MEFs and its viability wasn＇t affected. However, the organization of F-actin and microtubule was disturbed and MTOC was separated from nucleus which eventually inhibited cell migration. Conclusion Nesprin-1 regulated the stiffness of cytoskeleton and polarization of MTOC by the connection of nucleus and cytoskeleton to affect cell migration.
出处
《中国处方药》
2014年第10期1-3,共3页
Journal of China Prescription Drug
基金
国家自然科学基金(81102022)
教育部高等学校博士学科点专项科研基金(新教师类)(20110171120082)
高校基本科研业务费中山大学青年教师培育项目(14ykpy20)
