外源性重组S100A6蛋白对5株转化细胞系的增殖、迁移能力及β-catenin水平的影响  

Effects of exogenetic recombinant protein S100A6 on proliferation and migration of 5 transformed cell lines and β-catenin expression level

作　　者：马闻[1] 李星星[1] 王胜[2] 游莉[3] 郭元元[3] 徐兰兰[3] 周兰[3] 

机构地区：[1]重庆医科大学生物医学工程系 [2]重庆医科大学实验动物中心 [3]重庆医科大学医学检验系临床检验诊断学、教育部重点实验室,重庆400016

出　　处：《重庆医科大学学报》2010年第9期1321-1326,共6页Journal of Chongqing Medical University

摘　　要：目的:探讨S100A6对5种转化细胞系株(HEK293、9HTE、CHO、MEF和C3H10T1/2)的增殖、迁移能力和β-catenin水平的影响。方法:通过转化质粒pMoluc-GST-hS100A6到大肠杆菌BL21,经IPTG诱导表达重组S100A6蛋白,为后续实验提供材料;MTT法检测细胞增殖,细胞划痕和Transwell法检测细胞迁移,免疫细胞化学法及Western-blotting检测β-catenin表达水平。结果:①通过IPTG诱导表达,从大肠杆菌中收获了高纯度重组S100A6(GST-S100A6);②外源性重组S100A6对作为阳性对照的肿瘤细胞系——人骨肉瘤细胞143B的增殖及迁移均有抑制作用(P<0.05),而对5株转化细胞系的增殖及迁移均无显著影响(P>0.05);③免疫细胞化学法和Western-blotting一致揭示重组S100A6可上调5株转化细胞系的β-catenin水平(P<0.05)。结论:S100A6对5株转化细胞系的增殖、迁移无明显影响,但可上调此5株转化细胞系Wnt/β-catenin水平。Objective：This study aimed to investigate effects of exogenetic recombinant protein S100A6 on 5 transformed cells（HEK293,9HTE,CHO,MEF and C3H10T1/2） and their β-catenin expression level.Methods：Plasmid pMoluc-GST-hS100A6 was transformed into E coli.BL21,in which S100A6 can be induced to express by IPTG;Cell proliferation was measured by MTT,cell migration was measured by cell scratch assay and Transwell assay and levels of β-catenin were measured by ICC and Western-blotting.Results：① High purity recombinant S100A6（GST-S100A6） was obtained from E Coli via induction of IPTG induction.②Exogenetic S100A6 can not only inhibit 143B cell lines（positive control） proliferation but also the migration（P0.05）.However,effects of different concentration of S100A6 cast on proliferation（P0.05） and migration（P0.05） of these 5 transformed cell lines were not statistically significant.③ S100A6 can up-regulate β-catenin level in these 5 transformed cell lines significantly（P0.05）.Conclusion：Effects of S100A6 on these 5 transformed cell lines were not significant neither on proliferation nor on migration,but it can up-regulate their β-catenin levels.

关 键 词：S100A6 转化细胞系 细胞增殖 细胞迁移 Β-CATENIN 

分 类 号：R730.5[医药卫生—肿瘤]