【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。更多还原

【Abstract】 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(P<0.05).However,effects of different concentration of S100A6 cast on proliferation(P>0.05) and migration(P>0.05) 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(P<0.05).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.更多还原