IL-4激活的巨噬细胞通过分泌CCL18促进Hela细胞侵袭和迁移

IL-4 Activated Macrophages Promote Hela Cell Migration and Invasion by Producing CCL18

【目的】研究IL-4激活的巨噬细胞对宫颈癌Hela细胞的侵袭、转移作用及其机制。【方法】细胞形态观察、流式细胞术及ELISA检测巨噬细胞的激活状态。Transwell法观察Hela细胞的侵袭转移能力。Western blot检测NF-κB信号蛋白的激活状况及转移相关基因的表达。【结果】(1)IL-4激活的巨噬细胞表现为免疫抑制的M2型表型;(2)IL-4激活的巨噬细胞显著促进Hela细胞的侵袭、迁移,而未激活的巨噬细胞没有这种能力(P<0.001);(3)IL-4激活的巨噬细胞上清中加入CCL18中和抗体而不是同型Ig G对照能削弱M2型巨噬细胞的作用(P<0.001);(4)IL-4激活的巨噬细胞上清能促进共培养肿瘤细胞中NF-κB信号蛋白的激活和vimentin的表达;(5)直接应用CCL18也可以促进Hela细胞的侵袭转移,NF-κB信号蛋白的激活和vimentin的表达。【结论】IL-4激活的巨噬细胞可以促进宫颈癌细胞的侵袭转移,其作用机制为释放CCL18作用于肿瘤细胞的NF-κB通路,进而促进转移相关基因的表达。

【Objective】To investigate the roles of IL-4 activated macrophages on Hela cells migration and invasion and the underlying mechanism.【Methods】The phenotype of IL-4 activated macrophage was assayed by morphology,flow cytometry and Elisa. The ability of Hela cell migration and invasion was evaluated by transwell assay. The signaling proteins expression was displayed by Western blot.【Results】Il-4 activated macrophage had the phenotype of CD206high/HLA-DRlow,and highly expressed IL-10 and CCL18,and decreased the expression of IL-12. The activated macrophages significantly promoted the migration and invasion of Hela cells,while the unactivated macrophages did not had this effect. Applying anti-CCL18 antibody but not isotype IgG in the supernatant of IL-4 activated macrophage inhibited Hela cells motility. Furthermore,activated macrophages phosphorylated IKK and IκB and induced the translocation of p65 to nucleus,eventually increased the expression of vimentin. Treatment of Hela cells with CCL18 also enhanced tumor cells migration and invasion,as well as the NF- κB activation and vimentin expression.【Conclusion】IL-4 activated macrophages promoted Hela cells motility by secreting CCL18 and the following activation of NF- κB signaling proteins.