多核人宫颈癌Hela细胞抗失巢凋亡的研究  

Research on anoikis resistance of multinucleated cervical carcinoma Hela cells

作　　者：胡赛赛 胡田勇[2] 陈艳艳[2] 罗纯宾 姚志彬[1] 郑小娜 HU Sai-sai;HU Tian-yong;CHEN Yan-yan;LUO Chun-bin;YAO Zhi-bin;ZHENG Xiao-na(Department of Anatomy. Zhongshan School of Medicine, Sun Yat- sen University, Guangzhou 510080, Guangdong, China)

机构地区：[1]中山大学中山医学院人体解剖学教研室,广东广州510080 [2]深圳市龙岗中心医院耳鼻咽喉科医院深圳市耳鼻咽喉研究所,广东深圳518172 [3]南方医科大学中西医结合医院,广东广州510315

出　　处：《解剖学研究》2018年第2期90-94,F0004,共6页Anatomy Research

基　　金：广东省科技发展专项资金项目(2017A030310231);深圳市重点实验室基金项目(ZDSYS201506050935272);龙岗区科技发展资金项目(20160607142024828)

摘　　要：目的探讨多核癌细胞与细胞外基质脱离联系即"失巢"后细胞核状态的改变及其发生机理。方法通过从人宫颈癌Hela细胞中筛选出多核细胞,利用荧光染色对F-actin和细胞核进行双重标记以检测细胞核分裂以及细胞质分裂是否发生异常。采用聚羟乙基异丁烯酸(Poly-HEMA)阻断细胞与细胞外基质联系的方法,模拟"失巢"生长环境,用Annexin V-PI双染法检测多核细胞发生失巢凋亡的情况。然后使失巢后的多核细胞重新贴壁,观察细胞核形态以及个数的变化。通过三重荧光染色法观察不同细胞核个数的多核细胞其线粒体形态以及与F-actin共定位情况的差异。结果细胞质分裂受阻与多核Hela细胞的产生有关;细胞核个数较多的细胞抗失巢凋亡能力较低,这可能是由F-actin构成的细胞骨架系统在运输线粒体的过程中发生异常而使线粒体在细胞核边缘成簇富集造成的。结论多核癌细胞的抗失巢凋亡能力与其细胞核、细胞骨架以及线粒体的状态有关。Objective To investigat whether and why the nuclei of multinucleated tumor cells can change after matrix- detachment. Methods Multinueleated tumor ceils were collected from human cervical carcinoma Hela cells. By double fluorescence labeling of F-actin and nuclear, whether the abnormity of karyonitosis or eytokines is related with the occurence of multinucleated cells was determined. Polyhydroxyethylmethacrylate （Poly-HEMA） was used to establish the detachment condition by cell-contact blocking. Anoikis of multinueleated cells was measured by Annexin V-PI staining. Multinueleated cells were re-attached and then the morphological change of their nuclei was detected. Further, by triple staining, differences of the morphology of mitochondria and its colocation with F-aetin were distinguished in different types of muhinucleated ceils. Results Muhinucleated cells were resulted from inhibition of cytokinesis. Of note, muhinucleated cells with more number of nuclei were easier to undergo anoikis, which may be caused by peri-nuclear mitochondria clustering that is due to disruption of mitochondrial migration system which is dependent on F-actin eytoskeleton. Conclusion The anoikis-re- sistance capacity of multinueleated tumor cells was related with the status of the nuclei, cytoskeleton and mitochondria.

关 键 词：多核细胞 HELA细胞 失巢凋亡 宫颈癌 

分 类 号：R737.33[医药卫生—肿瘤]