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目的 :探讨他莫昔芬(tamoxifen,TAM)通过雌激素G蛋白偶联受体30(estrogen G-protein-coupled receptor 30,GPR30)介导肿瘤相关成纤维细胞(cancer-associated i broblast,CAF)外分泌CXC型趋化因子配体16(CXCchemokine ligand 16,CXCL16)对乳腺癌MCF-7细胞迁移和侵袭的影响。方法 :TAM和TAM联合GPR30特异性抑制剂G15分别处理CAF后,通过基因芯片检测2组细胞差异表达的基因,并从中挑选出CXCL16,应用实时荧光定量PCR和蛋白质印迹法进行验证,应用ELISA法检测细胞上清液中CXCL16的浓度。用TAM和TAM联合G15处理后的CAF上清液、不做任何处理的CAF上清液、添加CXCL16或CXCL16中和抗体的培养液分别培养MCF-7细胞,采用MTT法、划痕愈合实验和Transwell小室法检测细胞的增殖、迁移和侵袭能力。结果 :与TAM联合G15处理组相比,TAM处理组CAF中2 358个基因表达上调,2 178个基因表达下调;TAM处理组CAF中CXCL16 m RNA和蛋白的表达水平及上清液中CXCL16的浓度均显著上调(P值均<0.01),且该效应可被G15阻断(P值均<0.01)。TAM处理组CAF上清液和添加CXCL16培养液培养的MCF-7细胞,其迁移和侵袭能力均增强(P值均<0.05),而增殖能力则无显著变化(P值均>0.05)。结论 :CAF经TAM处理后CXCL16分泌增多,该效应由GPR30介导,而外分泌的CXCL16可促进乳腺癌MCF-7细胞的迁移和侵袭能力。
Objective: To investigate the effect of tamoxifen on the expression of cancer-associated i broblast (CAF) in estrogen G-protein-coupled receptor 30 (GPR30) Effects of exocrine CXC chemokine ligand 16 (CXCL16) on the migration and invasion of breast cancer MCF-7 cells. Methods: TACE, TAM and G15 combined with GPR30 specific inhibitor G15 were used to detect the differentially expressed genes in the two groups by gene chip. CXCL16 was selected from them and verified by real-time fluorescence quantitative PCR and Western blotting. The concentration of CXCL16 in the cell supernatant was measured. MCF-7 cells were cultured in CAF supernatants treated with TAM and TAM combined with G15, without any treatment, and cultured in neutralizing medium supplemented with CXCL16 or CXCL16. MTT assay, wound healing assay and Transwell chamber assay of cell proliferation, migration and invasion. Results: Compared with TAM combined with G15 treatment group, 2 358 genes were up-regulated and 2 178 genes were down-regulated in CAF of TAM treatment group; CXCL16 m RNA and protein expression in CAF of TAM treatment group and CXCL16 in supernatant (All P <0.01), and this effect could be blocked by G15 (all P <0.01). The migration and invasion ability of CAF supernatants and MCF-7 cells cultured in CXCL16 medium were enhanced (all P <0.05), while the proliferative capacity did not change significantly (all P> 0.05). CONCLUSIONS: CXCL16 secretion increased after CAF treatment with TAM, which is mediated by GPR30, while exocrine CXCL16 promotes the migration and invasion of breast cancer MCF-7 cells.