人结直肠癌细胞诱导成纤维细胞分泌的基质细胞衍生因子1(SDF-1)促进癌细胞的迁移

Human colorectal cancer cells induce fibroblasts to secrete stromal cell-derived factor 1(SDF-1)to stimulate cancer cell migration

目的研究HCT-116结直肠癌细胞对癌旁正常成纤维细胞(NF)的炎性诱导作用及癌相关成纤维细胞(CAF)对癌细胞迁移的影响和作用机制。方法利用酶消化法分离NF和CAF;通过空间基因表达数据集下载并分析转化生长因子β(TGF-β)、肿瘤坏死因子α(TNF-α)、CXC趋化因子配体2(CXCL2)、白细胞介素1β(IL-1β)和基质细胞衍生因子1(SDF-1)等炎性因子在人结直肠癌组织的空间表达水平;通过体外直接或间接共培养体系研究成纤维细胞和癌细胞之间的相互作用;流式细胞术检测成纤维细胞中成纤维细胞激活蛋白(FAP)和α平滑肌肌动蛋白(α-SMA)的表达;ELISA检测培养基中TGF-β、TNF-α、CXCL2、IL-1β和SDF-1的分泌水平。设置HCT-116细胞和NF共培养组、HCT-116细胞和CAF共培养组、加入SDF-1抑制剂AMD3100的HCT-116细胞和CAF共培养组,采用细胞划痕实验和Transwell^(TM)迁移实验检测癌细胞的迁移能力。结果CAF比NF表达更高水平的FAP和α-SMA,并且HCT-116癌细胞可诱导NF向CAF转化;与NF相比较,CAF可分泌更高水平的TGF-β、TNF-α、CXCL2、IL-1β和SDF-1等炎性因子,但在共培养条件下,癌细胞只能诱导NF分泌SDF-1,对TGF-β、TNF-α、CXCL2和IL-1β的分泌无明显诱导作用;与NF相比较,CAF明显促进癌细胞的迁移,并且这种促进作用可被SDF-1抑制剂AMD3100减弱。结论结直肠癌细胞可以诱导癌旁NF向CAF的转化,而CAF通过分泌SDF-1促进癌细胞的迁移。

Objective To investigate the inflammatory induction effect of HCT-116 colorectal cancer cells on normal fibroblasts(NFs)extracted from para-cancerous tissues and the effect of cancer associated fibroblasts(CAFs)on cancer cell migration and its potential mechanism.Methods The enzyme digestive method was used to extract NFs and CAFs from tissues.Spatial Gene Expression Dataset was downloaded to analyze transcriptional expression levels of inflammatory factors including transforming growth factorβ(TGF-β),tumor necrosis factor-α(TNF-α),C-X-C motif chemokine ligand 2(CXCL2),interleukin-1β(IL-1β),and stromal cell-derived factor-1(SDF-1).In vitro direct and indirect co-culture models were used to study the interaction between fibroblasts and cancer cells.Flow cytometry was used to detect expressions of fibroblast activated protein(FAP)andα-smooth muscle actin(α-SMA).Inflammatory factors TGF-β,TNF-α,CXCL2,IL-1β,and SDF-1 secreted in medium were measured by ELISA.Three co-culture groups were set up in which HCT-116 cells were co-cultured respectively with NFs,CAFs,and CAFs pretreated with AMD3100,an inhibitor of SDF-1.Cell migration ability was evaluated by the cell scratch-wound assay and the Transwell^(TM) migration assay.Results CAFs expressed higher levels of FAP andα-SMA than NFs,and HCT-116 cancer cells induced the transformation of NFs to CAFs.Compared to NFs,CAFs secreted higher levels of inflammatory factors TGF-β,TNF-α,CXCL2,IL-1β,and SDF-1,while in the in vitroco-culture models cancer cells induced the secretion of SDF-1 but had no effect on secretions of TGF-β,TNF-α,CXCL2,and IL-1βfrom NFs.CAFs significantly stimulated cancer cell migration compared to NFs,which was weakened by AMD3100,an inhibitor of SDF-1.Conclusion Colorectal cancer cells induce the transformation of NFs to CAFs,and CAFs stimulated cancer cell migration with the SDF-1 secreted.