中华皮肤科杂志

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内皮素3上调黑素瘤细胞株A375表达骨桥蛋白的研究

黄长征1, 田进1, 陶娟1, 刘业强1, 李延1, 杨凌云1, 张晶1, 陈思远1, 钱悦1, 沈关心2, 李家文1, 涂亚庭1   

  1. 1. 华中科技大学同济医学院附属协和医院皮肤科, 武汉, 430022;
    2. 华中科技大学同济医学院免疫学系
  • 收稿日期:2006-12-01 出版日期:2007-10-15 发布日期:2007-10-15
  • 基金资助:
    国家自然科学基金(30671891)

Endothelin 3 upregulates the expression of osteopontin in human melanoma cells A375

HUANG Chang-zheng1, TIAN Jin1, TAO Juan1, LIU Ye-qiang1, LI Yan1, YANG Ling-yun1, ZHANG Jing1, CHEN Si-yuan1, QIAN Yue1, SHEN Guan-xin2, LI Jia-wen1, TU Ya-ting1   

  1. Department of Dermatology, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430022, China
  • Received:2006-12-01 Online:2007-10-15 Published:2007-10-15

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摘要： 目的 探讨内皮素受体(ETR)-B拮抗剂BQ788和磷脂酰肌醇-3激酶(PI3K)抑制剂LY294002对黑素瘤细胞株A375骨桥蛋白基因的表达。方法 对照组在人A375细胞培养体系中加入100 nmol/L内皮素3,试验组除加入100nmol/L内皮素3外,分别加入1μmol/L BQ788和20μmol/L、50μmol/L LY294002,采用RT-PCR和蛋白质印迹方法检测骨桥蛋白的mRNA和蛋白表达。结果 ETR-B拮抗剂BQ788和PI3K抑制剂LY294002均能显著减少人恶性黑素瘤细胞株A375中骨桥蛋白mRNA和蛋白的表达,与对照组比较,差异均有统计学意义(P<0.05)。高浓度较低浓度LY294002对骨桥蛋白表达的抑制作用更明显(P<0.05)。结论 内皮素3/ETR-B能通过激活PI3K信号通路上调骨桥蛋白的表达。

关键词: 黑色素瘤, 受体, 内皮素B, 内皮缩血管肽, A375, 骨桥蛋白

Abstract: Objective To investigate the effect of endothelin receptor B(ETR-B)antagonist, BQ788,and phosphatidylinositol-3 kinase(PI3K)inhibitor,LY294002,on the gene expression of osteopontin (OPN)in human melanoma cells A375.Methods A375 cells were cultured,and pretreated with 1μmol/L BQ788,20μmol/L and 50μmol/L LY294002,respectively,for 1 h before the addition of 100 nmol/L endothelin 3(ET-3)into the medium.Cells treated only with ET-3 served as the control.The mRNA and protein expres-sions of OPN were detected by RT-PCR and Western blot respectively.Results The mRNA and protein expressions of OPN were significantly decreased in A375 cells treated with BQ788,and in those treated with LY294002,as compared to those in the control cells(all P<0.05).The inhibitory effect of LY294002 at higher concentrations was more significant than that at lower concentrations(P<0.05).Conclusion ET3/ETR-B axis could upregulate the expression of OPN via activating the PI3K pathway in A375 cells.

Key words: Melanoma, Receptor, endothelin B, Endothelin-3, A375, Osteopontin