融合蛋白GST-hS100A2对人结肠癌细胞HCT116和SW480的抑制作用

Inhibitory Effect of GST-hS100A2 Fusion Protein on Human Colorectal Carcinoma HCT116 and SW480 Cells

目的探讨GST-人S100A2融合蛋白(GST-hS100A2)对人结肠癌细胞HCT116和SW480的抑制作用。方法将重组质粒pGST-moluc-hS100A2和pGST-moluc转化大肠杆菌BL21(DE3),IPTG诱导表达,Western blot鉴定后采用谷胱甘肽-琼脂糖4B球珠分离纯化。以制备的GST-hS100A2融合蛋白分别处理HCT116和SW480细胞,以GST处理的细胞和空白细胞作为对照,分别用MTT法检测细胞增殖活力,Hochest33258染色检测细胞凋亡,流式细胞术检测细胞周期,划痕愈合法检测细胞迁移能力,Transwell小室试验检测细胞侵袭能力。结果融合蛋白GST-hS100A2能以剂量依赖和时间依赖的方式抑制HCT116和SW480细胞的增殖,并可明显促进两种细胞凋亡,阻滞HCT116细胞周期,抑制HCT116细胞迁移和两种细胞的侵袭能力。结论 GST-hS100A2对人结肠癌HCT116和SW480细胞具有一定的抑制作用,有可能成为结肠癌分子治疗的新靶标。

Objective To investigate the inhibitory effect of fusion protein GST-hS100A2 on human colorectal carcinoma HCT-116 and SW480 cells.Methods Recombinant plasmids pGST-moluc-hS100A2 and pGST-moluc were transformed to E.coli BL21（DE3） for expression under induction of IPTG.The expressed products were identified by Western blot,then purified with Glutathion-Sepharose 4B beads.HCT116 and SW480 cells were treated with the prepared GST-hS100A2 fusion protein,using those treated with GST and those untreated as controls,and determined for proliferative activity by MTT method,for apoptosis by Hochest33258 staining,for cell cycle by flow cytometry,for migration ability by wound healing assay,and for invasion ability by Transwell test.Results GST-hS100A2 showed dose-and time-dependent inhibitory effect on the proliferation of HCT116 and SW480 cells.Meanwhile,the fusion protein significantly promoted the apoptosis and inhibited the invasion ability of both HCT116 and SW480 cells,while arrested the cell cycle and inhibited the migration of HCT116 cells.Conclusion GST-hS100A2 showed a certain inhibitory effect on HCT116 and SW480 cells and might be used as a new target for molecular therapy of colorectal carcinoma.