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中国癌症防治杂志

中国癌症防治杂志 ›› 2017, Vol. 9 ›› Issue (1): 40-45.doi: 10.3969/j.issn.1674-5671.2017.01.08

• 基础研究 • 上一篇    下一篇

RSK4与 TRAF4在不同乳腺细胞中的表达及分子作用通路研究

  

  1. 广西医科大学附属肿瘤医院乳腺一区；广西医科大学研究生院
  • 出版日期:2017-02-25 发布日期:2017-03-24
  • 通讯作者: 杨华伟。 E-mail:lordyhw@163.com
  • 作者简介:【共同第一作者】 朱佳
  • 基金资助:
    国家自然科学基金资助项目（81260394）

Expression of RSK4 and TRAF4 in different breast cell lines and cross-talk mechanism

  1. Department of Breast Surgery，Affiliated Tumor Hospital of Guangxi Medical University；Graduate School of Guangxi Medical University
  • Online:2017-02-25 Published:2017-03-24
  • Contact: Yang Huawei. E-mail:lordyhw@163.com

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摘要：

目的 研究抑癌基因p90核糖体S6蛋白激酶4（p90 ribosomal protein S6 kinase4，RSK4）相互作用蛋白与肿瘤坏死因子受体相关因子4（tumor necrosis factor receptor associated factor 4 ，TRAF4）在不同乳腺细胞株中的表达，探讨RSK4-TRAF4相互作用蛋白所在分子作用通路对乳腺癌侵袭转移的影响。方法 采用实时荧光定量PCR 法检测 RSK4和TRAF4在 HBL-100正常乳腺细胞株和MCF-7、Her-2阳性型、MDA-MB-231人乳腺癌细胞株中的表达情况。用蛋白质免疫印迹法（Western-blot）检测RSK4和TRAF4以及MAPK通路下游蛋白ERK1/2在HBL-100、MCF-7、Her-2阳性型、MDA-MB-231 4种乳腺细胞株中的表达。结果 RSK4 mRNA 、TRAF4 mRNA 在4种细胞中均表达，RSK4 mRNA在HBL-100表达量最高，在MCF-7、Her-2阳性型、MDA-MB-231 细胞中含量依次降低；TRAF4 mRNA在4种细胞中的表达量由低到高依次为HBL-100、MCF-7、Her-2阳性型、MDA-MB-231 细胞，RSK4 mRNA 、TRAF4 mRNA 在4种细胞中两两比较，差异有统计学意义（P<0.05）。RSK4与TRAF4在4种细胞中蛋白表达水平与mRNA表达水平相一致，具有高度相关性（rRSK4=0.92,P< 0.01；rTRAF4=0.82,P< 0.01）；ERK1/2蛋白表达量由高到低依次为MDA-MB-231、Her-2阳性型、MCF-7、HBL-100。在mRNA转录水平，RSK4和TRAF4 表现出负相关（r1=-0.81,P< 0.01）；在蛋白翻译水平，RSK4和TRAF4 同样呈现出高度负相关（r2=-0.84，P< 0.01）。结论 RSK4和TRAF4具有较高的负相关性，RSK4与TRAF4相互作用后可能通过MAPKs下游信号通路ERK起分子调控作用。

关键词: 乳腺肿瘤, 人乳腺癌细胞株, p90核糖体S6蛋白激酶4, 肿瘤坏死因子受体相关因子4, ERK信号通路

Abstract:

Objective To study the expression of tumor suppressor gene p90 ribosomal S6 kinase 4（RSK4） and the interacting protein tumor necrosis factor receptor-associated factor 4（TRAF4） in different human breast cancer cell lines，and to study the potential role of these proteins in breast cancer invasion and metastasis. Methods Real-time fluorescence quantitative PCR （RT-PCR） was used to detect the mRNA expression of TRAF4 and RSK4 in four human breast cancer cell lines （HBL-100，MCF-7，Her-2-positive type，MDA-MB-231）. Western blotting was used to detect the expression of RSK4，TRAF4 and MAPK pathway proteins downstream of ERK1/2 protein in the same four breast cancer lines. Results RSK4 and TRAF4 mRNAs were expressed in all four cell lines；the highest expression was observed for RSK4 mRNA in HBL-100 cells. Expression of RSK4 mRNA was slightly lower in MCF-7，Her-2-positive type and MDA-MB-231 cells. Expression of TRAF4 mRNA followed the trend: HBL-100P<0.05）. The levels of RSK4 and TRAF4 proteins in the four cell lines strongly correlated with the levels of mRNA（rRSK4=0.92，P<0.01；rTRAF4=0.82，P<0.01）；the expression of ERK1/2 protein in the four cell lines followed the trend: MDA-MB-231>Her-2-positive type>MCF-7> HBL-100. RSK4 mRNA levels correlated negatively with TRAF4 mRNA levels（r1=-0.81，P<0.01），and the same was observed for protein levels （r2=-0.84，P<0.01）. Conclusions RSK4 and TRAF4 expression negatively correlate with each other，which may be mediated by MAPKs downstream of the ERK pathway.

Key words: Breast neoplasms, Human breast cell lines, p90 ribosomal S6 kinase 4(RSK4), tumor necrosis factor receptor-associated factor 4 (TRAF4), ERK signaling pathway