论文部分内容阅读
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 GST-human S100A2 fusion protein (GST-hS100A2) on human colon cancer cell line HCT116 and SW480. Methods The recombinant plasmids pGST-moluc-hS100A2 and pGST-moluc were transformed into E. coli BL21 (DE3) and induced by IPTG. After identification by Western blot, they were isolated and purified by glutathione-Sepharose 4B beads. HCT116 and SW480 cells were treated with GST-hS100A2 fusion protein respectively. GST-treated cells and blank cells were used as controls. Cell viability was detected by MTT assay. Apoptosis was detected by Hochest33258 staining. Cell cycle was detected by flow cytometry. Scratch healing assay for cell migration, Transwell chamber assay to detect cell invasion. Results The fusion protein GST-hS100A2 could inhibit the proliferation of HCT116 and SW480 cells in a dose-dependent and time-dependent manner, and significantly promote the apoptosis of both cell lines, arrest the cell cycle of HCT116, inhibit the migration of HCT116 cells and the invasion ability of both cells . Conclusion GST-hS100A2 can inhibit human colon cancer HCT116 and SW480 cells and may become a new target of molecular therapy for colon cancer.