趋化因子受体5及其配体促进乳腺癌细胞的趋化和侵袭的研究

摘要

目的 探讨趋化因子受体5(CCR5)及其配体(RANTES)对乳腺癌细胞趋化活性和侵袭活性的促进作用.方法 用反转录聚合酶链反应、免疫组织化学法测定乳腺癌细胞株MCF-7的CCR5 mRNA及蛋白水平,RANTES的作用下,以无血清RPMI 1640培养液作为阴性对照组,含乳腺癌细胞的无血清RPMI1640培养液为对照组,加有CCR5mAb的乳腺癌细胞无血清RPMI 1640为实验组,通过趋化小室法检测MCF-7细胞的趋化活性和侵袭活性,并与CCR5的封闭进行对照研究.结果 迁移至多聚碳酸酯膜背面的乳腺癌细胞数明显多于相应的阴性对照组,实验组和对照组迁移至多聚碳酸酯膜背面的细胞分别为(73.0±9.8)、(7.6±1.5)个/HP,差异有统计学意义(=11.85,P＜0.01)；穿透Matrigel基质胶到达多聚碳酸酯膜背面的乳腺癌细胞数明显多于相应的阴性对照组,实验组和对照组迁移至多聚碳酸酯膜背面的细胞分别为(19.7±4.0)、(2.3±0.6)个/HP,差异有统计学意义(=7.37,P＜0.01).乳腺癌细胞株MCF-7有CCR5mRNA及蛋白质的表达,其配体RANTES对MCF-7细胞有明显的趋化活性和侵袭活性,CCR5的封闭也能抑制MCF-7细胞的这种趋化活性和侵袭活性.结论 乳腺癌细胞株MCF-7有CCR5的表达,且其表达能够促进MCF-7细胞的趋化和侵袭.%Objective To study the chemotaxis and invasion of breast cancer cells facilitated by chemokine receptor 5 (CCR5) and its ligand.Methods Reverse transcription-polymerase chain reaction (RT-PCR) and immunohistochemical staining were used todetect CCR5 mRNA and protein expression in breast cancer cells MCF-7 ;the chemotaxis and invasion of MCF-7 were performed by using Boyden chambers in presence of activation normal T-cell and secreted(RANTES) which is a ligand of CCR5.Results The number of cells migrating to the opposite side of polycarbonate membrane was significantly higher than that of negative control group,which was (73.0 ±9.8) and(7.6 ± 1.5)/HP respectively (t =11.85,P ＜ 0.01).The number of cells penetrating Matrigel gel and migrating to the opposite side of polycarbonate membrane was significantly higher than that of negative control group [(19.7 ± 4.0) and (2.3 ± 0.6)/HP respectively (t =7.37,P ＜ 0.01)].Breast cancer cells MCF-7 expressed CCR5 mRNA and protein,and RANTES-mediated chemotaxis and invasion could be checked in the breast cancer cells.Both chemotaxis and invasion could be blocked by neutralizing anti-CCR5 antibody.Conclusions Breast cancer cells MCF-7 expressed CCR5 and chemotaxis and invasion of MCF-7 can be facilitated by CCR5 expression.