选择性ER调节剂对骨肉瘤细胞增殖及CCL2表达的抑制作用研究

Inhibitory effects of antiestrogen on the proliferation and CCL2 expression of osteosarcoma cells

目的探讨选择性ER调节剂他莫昔芬对骨肉瘤细胞增殖及CC趋化因子配体(CCL)2表达的影响。方法检测骨肉瘤细胞(143B和U-2OS)及成骨细胞(hFOB1.19)中雌激素受体(ER)的表达量。采用细胞增殖实验和平板克隆形成实验观察他莫昔芬对骨肉瘤细胞增殖的影响。采用荧光定量聚合酶链反应(PCR)检测骨肉瘤细胞中炎症因子IL-6、IL-8、TGF-β、CCL2和CCL5基因的相对表达量。采用酶联免疫吸附试验(ELISA)检测他莫昔芬对骨肉瘤细胞CCL2表达的影响。结果2种骨肉瘤细胞(143B和U-2OS)的ER mRNA相对表达量均显著低于成骨细胞(hFOB1.19);他莫昔芬可显著抑制骨肉瘤细胞的增殖及炎症因子IL-6、IL-8、TGF-β、CCL2和CCL5基因的表达,其中对CCL2表达的抑制作用最为显著。结论他莫昔芬可抑制骨肉瘤细胞的增殖及CCL2的表达,提示其或可作为骨肉瘤的辅助化疗药物。

Objective To investigate the effects of antiestrogen(tamoxifen)on the proliferation and the expression level of CC chemokine ligand(CCL)2 of osteosarcoma cells.Methods The difference of estrogen receptor(ER)expression between osteosarcoma cells(143B,U-2OS)and osteoblast(hFOB1.19)was observed.CCK8 experiment and plate colony formation assay were used to observe the effects of tamoxifen on the proliferation of osteosarcoma cells.The relative expression levels of inflammatory factors,IL-6,IL-8,TGF-β,CCL2 and CCL5 in osteosarcoma cells were determined by fluorescence quantitative polymerase chain reaction(PCR).The effects of tamoxifen on CCL2 expression of osteosarcoma cells in cell culture medium were determined by enzyme-linked immunosorbent assay(ELISA).Results The expression level of ER mRNA in osteosarcoma cells(143B,U-2OS)was lower than that in osteoblast(hFOB1.19).Tamoxifen could inhibit the proliferation of osteosarcoma cells.Tamoxifen could inhibit the expressions of inflammatory factors,IL-6,IL-8,TGF-β,CCL2 and CCL5,especially the chemokine CCL2 in osteosarcoma cells.Conclusions Tamoxifen could inhibit the proliferation of osteosarcoma cells and the expression of CCL2,suggesting that it might be used as a adjuvant drug for postoperative chemotherapy of osteosarcoma.