肝再生磷酸酶-3促进结肠癌细胞分泌趋化因子26对肿瘤相关性巨噬细胞的趋化作用

摘要

Objective To investigate the phosphatase of regenerating liver-3 (PRL-3) promoting colon cancer cells LoVo to secrete chemokine 26 (CCL26),which enhances tumor-associated macrophages (TAMs) migration.Methods We used real-time quantitative reverse transcriptase-polymerase chain reaction (RT-qPCR),and enzyme linked immunosorbent assay (ELISA) to detect the mRNA and protein expression of CCL26,and the protein expression of CCL26 after treatment of LoVo cells with nuclear factor-κB (NF-κB) inhibitor,BAY 11-7082 (5,10,and 15 mol/L).Invasion assays were applied to determine the effect of CCL26 on the ability of PRL-3 promoting migration of TAMs.Results RT-qPCR showed that the CCL26 mRNA expression of LoVo-P was higher than LoVo-C [(46 ± 5) fold].ELISA displayed that the protein level of CCL26 of LoVo-P was higher than LoVo-C,and it was attenuated in a concentration-dependent manner by treating LoVo-P with the NF-κB inhibitor,BAY 11-7082 (5,10,and 15 mol/L).Transwell invasion assays showed that the migration of TAMs was enhanced when TAMs were cocultured with LoVo-P cells,and it can be inhibited by NF-κB inhibitor,BAY 11-7082.Also the migration of TAMs was enhanced in a concentration-dependent manner when TAMs were cocultured with different concentrations of CCL26 (P ＜ 0.01).Conclusion PRL-3 promotes colon cancer cells to secrete chemokine CCL26,which can enhance TAMs migration.%目的 观察肝再生磷酸酶-3(PRL-3)促进结肠癌细胞LoVo分泌趋化因子26(CCL26),对肿瘤相关性巨噬细胞的趋化、聚集作用.方法 实时定量反转录聚合酶链反应(RT-qPCR)检测CCL26 mRNA水平表达差异;酶联免疫吸附试验(ELISA)检测CCL26蛋白水平表达差异,检测添加核因子κB(NF-κB)抑制剂BAY 11-7082(5、10、15 mol/L)后,CCL26蛋白水平;Transwell迁移实验检测LoVo-P、LoVo-C与TAM共培养24h后,对TAM迁移作用.结果 RT-qPCR检测结果显示,LoVo-P对比LoVo-C,CCL26升高(46±5)倍,ELISA检测CCL26蛋白水平,LoVo-P为(91±5) ng/L,LoVo-C为(61 ±3) ng/L,LoVo-P加入核因子(NF)-κB抑制剂BAY 11-7082(5、10、15 mol/L)后,CCL26蛋白水平呈浓度梯度降低(P＜0.01);Transwell迁移实验结果显示,LoVo-P对比LoVo-C明显对TAM有趋化作用,加入NF-κB抑制剂BAY 11-7082后,迁移作用明显减弱,不同浓度的CCL26(1、10、100 μg/L),能对TAM的趋化作用呈浓度梯度增高(P＜0.01).结论 PRL-3能促进结肠癌LoVo细胞分泌CCL26,通过CCL26可以诱导TAM趋化、聚集.