前列腺癌相关成纤维细胞通过分泌趋化因子C-C基元配体15促进单核细胞迁移

Fibroblasts associated with prostate cancer promote monocyte migration by secreting c-c motif chemokine ligand 15

目的:探究前列腺癌微环境中趋化因子C-C基元配体15(CCL15)的表达、来源以及对单核细胞迁移能力的影响。方法:对正常前列腺组织和前列腺癌组织进行CCL15免疫荧光染色,使用免疫组化和免疫荧光双染实验探究CCL15的主要来源细胞。从新鲜的正常前列腺组织和前列腺癌组织中分离成纤维细胞并进行原代培养,通过实时定量PCR、免疫印迹实验和ELISA实验检测成纤维细胞中α-平滑肌肌动蛋白(α-SMA)、成纤维细胞活化蛋白(FAP)、波形蛋白(Vimentin)和CCL15的表达情况。利用慢病毒转染敲低CCL15,探究敲低CCL15后成纤维细胞条件培养基对单核细胞迁移能力的影响。进行多色免疫荧光染色检测前列腺癌组织中CCL15与M2巨噬细胞的位置关系。结果:前列腺癌组织中CCL15的表达明显高于正常组织,且主要来源于表达α-SMA的成纤维细胞。与前列腺正常成纤维细胞(NF)相比,前列腺癌相关成纤维细胞(CAF)分泌CCL15的能力显著增强。敲低CCL15后,前列腺CAF分泌的CCL15明显减少,减弱了单核细胞的迁移能力。在前列腺癌组织中,CCL15高表达区域存在更多M2巨噬细胞。结论:前列腺CAF通过分泌CCL15促进单核细胞迁移,增加前列腺癌微环境中M2巨噬细胞的浸润。

Objective To investigate the expression and origin of c-c motif chemokine ligand 15(CCL15)in the microenvironment of prostate cancer,as well as its impact on monocyte migration ability.Methods CCL15 immunofluorescence staining was performed on normal prostate tissue and prostate cancer tissue,and immunohistochemical and immunofluorescence double staining experiments were used to explore the main source cells of CCL15.Isolate fibroblasts from fresh normal prostate tissue and prostate cancer tissue and perform primary culture.Detect fibroblasts through qPCR,Western Blotting and ELISA experiments expression ofα-SMA,FAP,Vimentin and CCL15.Using lentivirus transfection to knock down CCL15,investigate the effect of knocking down CCL15 on the migration ability of monocytes in fibroblast conditioned medium.Perform multi-color immunofluorescence staining to detect the positional relationship between CCL15 and M2 macrophages in prostate cancer tissue.Results The expression of CCL15 in prostate cancer tissue is significantly higher than that in normal tissue,and it mainly comes from its expression ofα-SMA.Compared with normal fibroblasts,prostate cancer associated fibroblasts significantly enhance their ability to secrete CCL15.After knocking down CCL15,the secretion of CCL15 by cancer associated fibroblast was significantly reduced,weakening the migration ability of monocytes.In prostate cancer tissue,there are more M2 macrophages in the high expression region of CCL15.Conclusion CAF promotes monocyte migration and increases M2 macrophage infiltration in the prostate cancer microenvironment by secreting CCL15.