PPAR-γ配体罗格列酮对人黑素瘤细胞体外趋化功能的影响及相关机制

Inhibitory effect of PPAR-γligand rosiglitazone on chemotaxis of melanoma cell in vitro and its mechanism

目的:探讨过氧化物酶体增殖物激活受体-γ(PPAR-γ)配体罗格列酮(rosiglitazone,RGZ)对人黑素瘤A375细胞株体外趋化功能的影响及机制。方法:体外培养人黑素瘤A375细胞;四甲基偶氮唑蓝(MTT)法检测RGZ对A375细胞的增殖抑制率;RT-PCR、Western-blot分别检测RGZ对A375细胞的趋化因子受体CXCR4 mRNA及CXCR4蛋白表达的影响;用Boyden小室研究RGZ对A375细胞体外趋化功能的影响。结果:MTT法检测结果显示在24 h干预点,浓度为10、25μmol/L时,RGZ的细胞毒性作用不明显(P>0.05);且RT-PCR、Western-Blot检测结果分别显示,RGZ能下调CXCR4mRNA及CXCR4蛋白的表达(P<0.01)。趋化实验显示,随着RGZ浓度的升高,穿过Boyden小室膜的A375细胞数显著减少(P<0.01)。结论:PPAR-γ配体RGZ能抑制人黑素瘤细胞的趋化功能,其机制可能与下调肿瘤细胞的CXCR4基因表达有关。

Objective： To explore the effect of rosiglitazone （RGZ）, a ligand of PPAR-γ, on chemotaxis of melanoma cell in vitro and its mechanism. Methods： Human melanoma cell A375 was selected and cultured in vitro. The growth inhibition rate of A375 cell was detected by M3T. The level of the expression of CXCR4-mRNA was determined by semi-quantitative RT- PCR, and the level of CXCR4 protein by Western blot. The chemotactic ability of A375 cells were determined by Boyden Chamber Assay. Results： Among the low concentration groups （10, 25μmol/L） at the point of 24 hours, the growth inhibition rate was not showed obvious significance （P〉0.05）. Meanwhile it can down-regulate the expression of CXCR4-mRNA and CX- CR4 protein （P〈0.01）. At last, RGZ can significantly reduce cell numbers of A375 through the Boyden booth （P〈 0.01）. Conclusion： By down-regulating the expression of CXCR4-mRNA and CXCR4 protein, PPAR-γ ligand rosiglitazone has a significant inhibitory effect on chemotaxis of human melanoma cell A375.