基础医学与临床 ›› 2011, Vol. 31 ›› Issue (4): 388-393.

• 研究论文 • 上一篇    下一篇

表皮生长因子促进电压门控钠通道Nav1.5表达及人乳腺癌细胞的侵袭

潘惠艳1,赵丽红2,代荫梅2,张卫华2,陈虹3,黄秉仁3   

  1. 1. 首都医科大学附属北京妇产医院
    2. 首都医科大学 附属北京妇产医院
    3. 中国医学科学院 基础医学研究所 北京协和医学院 基础学院 生物化学和分子生物学系
  • 收稿日期:2010-10-09 修回日期:2010-12-10 出版日期:2011-04-05 发布日期:2011-04-08
  • 通讯作者: 黄秉仁 E-mail:huangbr@public.bta.net.cn
  • 基金资助:
    院校级基金

Epidermal growth factor upregulates voltage-gated sodium channel/Nav1.5 expression and human breast cancer cell invasion

PAN Hui-yan 1,ZHAO Li-hong 2,DAI Yin-mei 2,ZHANG Wei-hua 2,CHEN Hong 2,HUANG Bing-ren 3   

  1. 1. Beijing Obstetrics and Gynecology Hospital, Capital Medical University
    2.
    3. Institute of Basic Medical Sciences, CAMS & PUMC
  • Received:2010-10-09 Revised:2010-12-10 Online:2011-04-05 Published:2011-04-08
  • Contact: HUANG Bing-ren E-mail:huangbr@public.bta.net.cn

摘要: 目的 探讨表皮生长因子(EGF)是否调节乳腺癌MDA-MB-231细胞电压门控钠通道(VGSC)Nav1.5亚型基因的表达及其可能的信号分子传道途径。方法 用Matrigel侵袭、免疫荧光定位、实时荧光定量RT-PCR(RFQ-PCR)和Western blot等方法,分别检测或探讨EGFR和Nav1.5蛋白在细胞中的表达和定位、EGF和Nav1.5对细胞侵袭的作用、EGF对Nav1.5mRNA和蛋白水平的影响以及PI3K在EGF增加侵袭中的作用。结果 MDA-MB-231细胞高表达EGF受体和Nav1.5蛋白,EGF增加细胞的侵袭达(51 ± 2.6)%,VGSC抑制剂Tetrodotoxin(TTX)10μmol/L阻滞EGF诱导的细胞侵袭(P﹤0.05);EGF增加Nav1.5mRNA水平为(128±4)倍(P﹤0.05), Nav1.5蛋白水平有(39±4)%上升(P﹤0.05)。PI3K抑制剂Wortmannin抑制EGF诱导的Nav1.5mRNA和蛋白水平的增加,亦抑制EGF诱导的细胞侵袭。结论 EGF上调乳腺癌MDA-MB-231细胞VGSC Nav1.5亚型mRNA和蛋白的表达,促使乳腺癌细胞的侵袭,PI3K参与EGF诱导的Nav1.5的表达和细胞的侵袭。

关键词: 表皮生长因子, 电压门控钠通道, 细胞信号, 侵袭, 乳腺癌

Abstract: Objective To investigate whether epidermal growth factor (EGF) regulates expression of voltage-gated sodium channel (VGSC) subtype of Nav1.5 as well as the possible intracellular pathway through which EGF may influences Nav1.5 expression. Methods Matrigel test, immunofluorescence, real-time fluorogentic quantitative PCR ( RFQ-PCR) and Western blot were used to investigate the level and intracellular distribution of EGF receptor (EGFR) , Nav1.5 protein, the effect of EGF on Nav1.5 mRNA expression and possible effect of PI3K on EGF enhancement of MDA-MB-231 cell invasion. Results MDA-MB-231 cells significantly expressed higher EGFR and Nav1.5 protein. EGF increased invasion of the cells approximately 51% ± 2.6% (P﹤0.05), and Tetrodotoxin (TTX) , the blocker of VGSC, suppressed the effect of EGF on the cell invasion (P﹤0.05) . The level of Nav1.5 mRNA and protein were increased by EGF treatment by 128 ± 4 times and 39% ± 4% (P﹤0.05 for both), respectively. Wortmannin , the inhibitor of PI3K, significantly reduced the EGF-induced increase of both Nav1.5mRNA and protein, and also decresed MDA-MB-231 cell invasion induced by EGF through Nav1.5. Conclusions EGF upregulated Nav1.5 mRNA and protein expression which promoted the invasion of strongly metastatic breast cancer MDA-MB-231 cells, and PI3K was found to be involved in the signaling cascade of EGF-induced VGSC/Nav1.5 expression.

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