不同亚型的半乳凝素-9对结肠癌LoVo细胞和内皮细胞体外黏附作用的影响

目的 研究不同亚型的半乳凝素-9对结肠癌LoVo细胞侵袭转移相关分子表达的调节作用,以及该调节作用对LoVo细胞与内皮细胞体外黏附作用的影响.方法 半乳凝素-9基因不同亚型的表达载体转染结肠癌LoVo细胞24 h后,分别用逆转录聚合酶链反应(RT-PCR)和Western blot方法检测整合素β1、钙黏着素E、选择素E、细胞间黏附分子-1、CD44和基质金属蛋白酶-9表达的变化.LoVo细胞分别在转染半乳凝素-9、转染半乳凝素-9+半乳凝素-9抗体、转染半乳凝素-9+选择素E抗体和转染半乳凝素-9+β-乳糖条件下进行体外LoVo细胞与内皮细胞的黏附实验.结果 半乳凝素-9L能下调选择素E mRNA和蛋白的表达,半乳凝素-9M和半乳凝素-9S能上调选择素E mRNA和蛋白的表达.体外黏附实验中,转染空载体、转染半乳凝素-9L、转染半乳凝素-9M和转染半乳凝素-9S组的平均荧光强度分别为0.90±0.20、0.94±0.24、1.60±0.11和1.45±0.13.半乳凝素-9M和半乳凝素-9S可促进kVo细胞黏附于内皮细胞(P<0.05),而半乳凝素-9抗体、选择素E抗体和β-乳糖均可抑制此作用.结论 3种不同亚型的半乳凝素-9通过不同的方式调节LoVo细胞选择素E mRNA和蛋白的表达,影响LoVo细胞与内皮细胞的体外黏附;3种不同亚型的半乳凝素-9在介导肿瘤细胞转移过程中的作用可能是不同的.

Galectin-9 isoforms influence the adhesion between colon carcinoma LoVo cells and human umbilical vein endothelial cells in vitro by regulating the expression of E-selectin in LoVo cells

Objective To study the regulatory effect of galectin-9 isoforms on some molecules involved in cell adhesion/invasion, and the influence of this regulation action on the adhesion between colon carcinoma LoVo cells and human umbilical vein endothelial cells (HUVECs) in vitro. Methods Various expression vectors of galectin-9 isoforms were transfected into LaVo cells. 24 h after transfection, the expression of intcgrin-βl, E-cadherin, E-selectin, ICAM-1, CD44 and MMP-9 was detected by RT-PCR and Western blot analysis. LoVo celI-HUVEC adhesion assay was performed under conditions of galectin-9 transfection, galectin-9 transfection+galectin-9 antibody, galectin-9 transfection+E-selectin antibody and galectin-9 transfection+β-lactose, respectively. Results Galectin-9L down-regulates the mRNA and protein levels of E-selectin while galectin-9M and galectin-9S up-regulate the expression of E-selectin. In LoVo celI-HUVEC adhesion assay, the average fluorescence intensity of vector transfection group, galectin-9L transfection group, galectin-gM transfection group and galectin-9S transfection group was 0.90±0.20, 0.94±0.24, 1.60±0.11 and 1.45±0.13, respectively, indicating that galectin-gM and galectin-9S facilitated the adherence of LoVo cells to HUVECs (P<0.05). E-selectin antibody, galectin-9 antibody or β-lactose inhibited that effect. Conclusion Galectin-9 isoforms regulate the E-selectin expression in LoVo cells differently and thus influence the adhesion between LoVo cells and HUVECs in vitro in different modes. The mechanisms through which galectin-9 isoforms participate in the metastasis process of colon cancer may not be the same.