S100A6 in the microenvironment directly and indirectly promotes migration of colorectal carcinoma LoVo cells / 肿瘤

ABSTRACT

Objective: To deeply investigate the effect of S100A6 in the microenvironment on migration of colorectal carcinoma LoVo cells, and to explore the possible molecular mechanism. Methods: The recombinant protein glutathione S-transferase (GST) and the fusion protein GST-human S100A6 (GST-hS1 00A6) were prepared and identifed. After the treatment with GST-hS1 00A6 (30 (μg/mL), the migration of LoVo cells was detected by wound healing assay, the expressions of total protein kinase B (PKB, also known as Akt) and phosphorylation of Akt (p-Akt) in LoVo cells were detected by Western blotting. After the treatment with GST-hS1 00A6 and phosphatidylinositol 3-kinase (PI3K)/Akt signaling pathway inhibitor LY294002 alone or in combination, the migration of LoVo cells was detected by wound healing assay. After the macrophages were treated with the conditioned medium of LoVo cells stimulated by GST-hS1 00A6 (named as CM-A6-LoVo), the expression levels of M2 phenotype marker CD206 and M1 phenotype marker inducible nitric oxide synthase (iNOS) in the macrophages were detected by real-time fuorescent quantitative PCR, and the migration ability of LoVo cells co-cultured with the macrophages was detected by wound healing assay. Results: GST-hS100A6 and GST protein (as a control) were successfully prepared. Compared with GST group, the wound healing rate of LoVo cells treated with GST-hS1 00A6 was significantly elevated (P 0.05), and the macrophages co-cultured with LoVo cells could significantly promote the migration of LoVo cells (P < 0.05). Conclusion: S100A6 in microenvironment can directly and indirectly promote the migration of colorectal carcinoma LoVo cells, which maybe related to the regulation of PI3K/Akt signaling pathway and the induction of macrophage polarization to M2 phenotype.