癌相关成纤维细胞来源的CCL7对三阴性乳腺癌细胞增殖与侵袭作用的研究

Effect of cancer-associated fibroblasts-derived CCL7 on proliferation and invasion of triple-negative breast cancer cells

目的 研究癌间质微环境中癌相关成纤维细胞(CAFs)来源的CCL7对三阴性乳腺癌(TNBC)细胞增殖与侵袭的作用.方法 采用实时定量PCR和Western Blot分别测定TNBC的CAFs与癌旁成纤维细胞中趋化因子配体7(CCL7)的mRNA及蛋白相对表达水平;采用酶联免疫吸附实验(ELISA)定量检测CAFs与癌旁成纤维细胞条件培养基中的CCL7水平,确定两种细胞的CCL7旁分泌水平;采用MTS和Transwell侵袭实验分别检测CCL7对TNBC细胞株MDA-MB-231增殖和侵袭的影响.结果 与癌旁成纤维细胞相比,CAFs中CCL7的mRNA及蛋白相对表达水平均明显升高,差异均具有统计学意义(均P<0.01),且CAFs的CCL7旁分泌水平也明显升高,差异具有统计学意义(P<0.01).MTS和Transwell侵袭实验结果显示,CCL7治疗组MDA-MB-231细胞的增殖率和侵袭能力均明显高于未治疗组,差异均具有统计学意义(P<0.05,P<0.01).结论 TNBC间质微环境中的CAFs可分泌更多的趋化因子CCL7,且CCL7可促进TNBC细胞的增殖和侵袭.

Objective To investigate the effect of cancer-associated fibroblasts (CAFs)-derived chemokine ligands 7 (CCL7) on the proliferation and invasion of triple-negative breast cancer (TNBC) cells. Methods The mRNA expression level and protein level of CCL7 in CAFs and paracancerous fibroblasts were detected by quantitative real-time polymerase chain reaction (qRT-PCR) and Western Blot respectively. To confirm the paracrine level of CCL7 in CAFs and paracancerous fibroblasts, the protein levels of CCL7 in the corresponding conditional medium were detected through enzyme-linked immunosorbent assay (ELISA). The effect of CCL7 on the proliferation and invasion of MDA-MB-231 (TNBC cell line) was investigated by MTS assay and Transwell assay, respectively. Results In comparison with paracancerous fibroblasts, the mRNA expression level and protein level of CCL7 in CAFs were significantly increased (both P<0.01). There was an obviously increase of paracrine level of CCL7 in CAFs-conditional medium (P<0.01). The MTS assay and Transwell assay results indicated that CCL7 was more able to promote the proliferation and invasion of MDA-MB-231. Conclusion CAFs in the TNBC stroma can produce more chemokine CCL7, and CCL7 can promote the proliferation and invasion of TNBC cells