孕激素对黑素瘤细胞抑制性配体分子表达影响的体外研究  

Effects of progesterone on immunosuppressive ligands expression in melanoma cells in vitro

作　　者：方险峰[1,2] 张序心[1] 李家文[2] 

机构地区：[1]广西中医学院附属瑞康医院皮肤科,广西南宁530011 [2]华中科技大学同济医学院附属协和医院皮肤科,湖北武汉430022

出　　处：《中华肿瘤防治杂志》2010年第15期1176-1179,共4页Chinese Journal of Cancer Prevention and Treatment

摘　　要：目的:探讨孕激素对黑素瘤细胞免疫抑制性分子表达影响及可能的信号传导机制。方法:体外培养黑素瘤细胞系A375及A875,并以不同浓度孕激素、孕激素受体拮抗剂RU486以及PI3-K/Akt抑制剂LY294002进行诱导。RT-PCR、ELISA或蛋白质印迹法分别检测IL-10、TGF-β1及HLA-GmRNA及蛋白表达以及PI3-K/Akt信号途径Akt磷酸化水平。结果:黑素瘤细胞系A375及A875均在mRNA及蛋白水平表达IL-10及TGF-β1。孕激素浓度≥1×10-7mol/L对2株细胞IL-10表达均有显著上调作用;对TGF-β1表达无明显影响;不能诱导HLA-G表达。Akt抑制剂LY294002可消除孕激素对IL-10表达上调效应,而RU486无拮抗效应。孕激素浓度为1×10-7mol/L时上调A375细胞Akt磷酸化水平。结论:高浓度孕激素可体外上调黑素瘤免疫抑制分子IL-10的表达,并与不依赖经典细胞核受体的PI3-K/Akt途径激活有关。OBJECTIVE：To explore the gene expression regulatory effects of progesterone on immunosuppressive molecules in malignant melanoma cells and the probable signal transduction pathway.METHODS：Cultured malignant melanoma cell lines A375 and A875 were treated with progesterone and its antagonist RU486 or PI3-K/Akt inhibitor LY294002.The expression of anti-inflammatory mediators,such as IL-10,TGF-β1 and HLA-G,were detected by RT-PCR and ELISA or Western blot assay.The activated PI3-K/Akt pathway was evaluated by the level of Akt phosphorylation with Western blot assay.RESULTS：Both A375 and A875 cells expressed TGF-β1 and IL-10 under serum-free culture conditions as measured by RT-PCR and ELISA.High concentration progesterone （≥1×10-7 mol/L） enhanced IL-10 expression both in A375 and A875 cells,but did not alter TGF-β1 and HLA-G expressions.LY294002 abrogated progesterone effects on IL-10 expression regulation,and progesterone antagonist RU486 did not antagonize this effects.High concentration progesterone increased the level of Akt phosphorylation in A375 cells.CONCLUSION：Progesterone stimulates PI3-K-dependent phosphorylation of Akt leading to the up-regulation of IL-10 gene expression in melanoma cells.

关 键 词：孕激素类 黑色素瘤 实验性 白细胞介素10 

分 类 号：R739.5[医药卫生—肿瘤]