中华皮肤科杂志 ›› 2009, Vol. 42 ›› Issue (8): 575-578.

• 论著 • 上一篇    下一篇

恶性黑素瘤细胞中的谷氨酸信号通路及其与细胞骨架蛋白分子的作用机制

卢平1,周育森2,陈万荣2,宋智琦3   

  1. 1. 大连市皮肤病医院
    2.
    3. 大连医科大学附属第一医院皮肤科
  • 收稿日期:2008-07-08 修回日期:2008-10-16 出版日期:2009-08-15 发布日期:2009-08-10
  • 通讯作者: 卢平

Glutamate receptor signaling pathway in melanoma cells and its effect on cytoskeleton protein

  • Received:2008-07-08 Revised:2008-10-16 Online:2009-08-15 Published:2009-08-10

摘要:

目的 探讨谷氨酸受体通路对恶性黑素瘤(恶黑)细胞树突形态以及细胞骨架蛋白的作用。方法 激光共聚焦显微镜及原子力显微镜分析微管相关蛋白2a(MAP2a)、离子型谷氨酸受体(NMDAR2A)、代谢型谷氨酸受体(mGluR1)在人侵袭性恶黑细胞中的亚细胞定位,以及NMDAR拮抗剂MK-801及mGluR1拮抗剂CPCCOEt对微管蛋白分布、细胞树突形态的作用。测定细胞生长曲线,观察MK-801及CPCCOEt对恶黑细胞增殖的影响。结果 研究发现,上述分子具有相似的亚细胞分布:均主要位于细胞树突内;MK-801及CPCCOEt均可使细胞树突内微管密度增加,使恶黑细胞树突化;且与MAP2a具有协同作用;MK-801及CPCCOEt均对恶黑细胞增殖具有显著抑制作用。结论 谷氨酸受体通路参与恶黑细胞树突发育的生理功能,谷氨酸受体拮抗剂可抑制恶黑细胞增殖。

关键词: 谷氨酸受体;细胞骨架蛋白;恶性黑素瘤

Abstract:

Objective To investigate the effects of glutamate receptor signaling on melanoma cell dendrite morphology and cytoskeleton protein. Methods A metastatic human malignant melanoma cell line WM451LU was cultured and transfected by recombinant adenovirus vector carrying a cDNA encoding microtubule-associated protein 2a(MAP2a). MK-801, an antagonist of N-methyl-D-aspartate receptor (NMDAR), and CPCCOEt, an antagonist of metabotropic glutamate receptor 1 (mGluR1), were used to treat transfected or untransfected WM451LU cells. Confocal microscopy and three dimensional atomic force microscopy were used to assess subcellular location of NMDAR2A, mGluR1 and MAP2a as well as the distribution of α-tubulin in and dendrite morphology of WM451LU cells. The proliferation of WM451LU cells was estimated by cell survival growth curve. Results Confocal laser microscopy revealed that NMDAR2A, mGluR1 and MAP2a were mainly co-localized in melanoma cell dendrites. Both MK-801 and CPCCOEt increased the density of microtubules in cell dendrites and dendritic branching of WM451LU cells, and both effects of MK-801 and CPCCOEt were enhanced by the expression of MAP2a. Furthermore, the proliferation of WM451LU cells was significantly inhibited by MK-801 of 100 μmol/L and CPCCOEt of 10 μmol/L. Conclusions In melanoma cells, glutamate receptors may participate in the development of dendrites, and anta- gonists of glutamate receptors could inhibit the proliferation of melanoma cells.