CXCL10/CCL2 信号通路介导肿瘤微环境中宫颈癌的发展

CXCL10/CCL2 signaling pathway mediates the development of cervical cancer in tumor microenvironment

摘要:

目的 探讨趋化因子（C-X-C 基序）配体 10（Chemokine（C-X-C motif） ligand 10，CXCL10）/C-C 基元配体 2（CC chemokine ligand 2，CCL2） 信号通路在肿瘤微环境（tumor microenvironment，TME）中宫颈癌（cervical cancer，CC）发展中的作用。方法 体外实验将宫颈癌细胞系 Hela 细胞分别用针对 CXCL10 基因的 shRNA（sh-CXCL10）和CXCL10 cDNA（CXCL10）转染处理，并通过 EdU 分析试验评估细胞增殖能力，和 Transwell 检测细胞侵袭能力。将 THP-1 细胞用佛波醇 12-肉豆蔻酸 13-乙酸酯（phorbol 12-myristate 13-acetate，PMA）诱导分化，并与转染慢病毒或质粒的 Hela 细胞以 1∶4 的比例组合共培养 48 h。通过流式细胞术检测 CD206+巨噬细胞数目和免疫印迹检测 STAT3/NF-κB 信号表达。结果 与 NC 组相比，与 THP-1 细胞共培养的 CXCL10 过表达的 Hela 细胞表现出增殖、迁移能力增强和侵袭细胞数增加（P<0.05），并且 CD206+巨噬细胞的比率显著增加（P<0.05），而与 THP-1细胞共培养的 CXCL10 敲低的 Hela 细胞则显示相反效果。与 NC 组相比，CXCL10 组共培养系统中 p-STAT3、p-NF-κB 表达和 CCL2 水平显著增加（P<0.05）。与 sh-NC相比，sh-CXCL10共培养系统中p-STAT3、p-NF-κB 表达和 CCL2 水平显著减少（P<0.05）。结论 CC 细胞中 CXCL10 上调促进 TME 中巨噬细胞 M2 极化，其机制可能与激活巨噬细胞的 STAT3/NF-κB/CCL2 信号传导途径有关，导致 CC 细胞增殖、迁移和侵袭能力增强。

Abstract:

Objective To investigate the role of C-X-C motif chemokine ligand 10（CXCL10）/C-C motif chemokine ligand 2（CCL2） signaling pathway in the development of cervical cancer（CC） in tumor microenvironment.Methods In vitro cervical cancer Hela cells were transfected with shRNA（sh-CXCL10） and CXCL10 cDNA（CXCL10） targeting CXCL10 gene，respectively.Cell proliferation was evaluated by EdU analysis and cell invasion was detected by Transwell assay.THP-1 cell differentiation was induced with phorbol 12-myristic acid 13-acetate（PMA）.The cells were co-cultured with Hela cells（5×10⁶ cells） transfected with the lentivirus or plasmid at a ratio of 1∶4 for 48 h.The number of CD206+ macrophages was determined by flow cytometry and STAT3/NF-κB signal expression was measured by Western blot.Results Compared with the NC group，Hela cells with CXCL10 over-expression co-cultured with THP-1 cells showed increased proliferation，migration，and invasion（P<0.05），and the ratio of CD206+ macrophages increased significantly（P<0.05）.Hela cells with CXCL10 knock-down co-cultured with THP-1 cells showed the opposite results.Compared with NC group，the CXCL10 co-culture system showed significant increases in the expression of p-STAT3，p-NF-κB，and CCL2（P<0.05）.Compared with the sh-NC group，the sh-CXCL10 co-culture system showed significant decreases in the expression of p-STAT3，p-NF-κB，and CCL2（P<0.05）.Conclusion Up-regulation of CXCL10 in CC cells promotes M2 polarization of macrophages in tumor microenvironment.The mechanism may be related to activation of STAT3/NF-κB/CCL2 signal transduction pathway in macrophages，which leads to the enhancement of proliferation，migration，and invasion of CC cells.