基础医学与临床 ›› 2015, Vol. 35 ›› Issue (10): 1382-1386.

• 研究论文 • 上一篇    下一篇

转染pcDNA3-TRAF4-DM-TRAF促进人乳腺癌细胞MDA-MB-231细胞增殖

张晓丽1,米小轶2   

  1. 1. 沈阳市妇婴医院
    2. 中国医科大学
  • 收稿日期:2015-04-23 修回日期:2015-07-02 出版日期:2015-10-05 发布日期:2015-09-30
  • 通讯作者: 米小轶 E-mail:xiaoyi_mi@163.com

Transfecting pcDNA3-TRAF4-DM-TRAF iimproves cell proliferation of human breast cancer cell MDA-MB-231

Xiao-Li ZHANG1,Xiao-Yi MI   

  • Received:2015-04-23 Revised:2015-07-02 Online:2015-10-05 Published:2015-09-30
  • Contact: Xiao-Yi MI E-mail:xiaoyi_mi@163.com

摘要: 目的 探讨TRAF4在人乳腺癌细胞高表达时对细胞增殖能力的影响。方法 本实验共分两组:实验组(在乳腺癌细胞MDA-MB-231中转染pcDNA3-TRAF4-DM-TRAF质粒)、对照组(转染pcDNA3空质粒),用细胞免疫荧光及免疫印迹法检测TRAF4的表达及定位,免疫印迹法检测p70S6K及S6蛋白磷酸化,用流式细胞术检测各期细胞比例,MTT方法检测增殖能力。结果 TRAF4在乳腺癌细胞MDA-MB-231细胞质及细胞核中均有表达,并且其在细胞核中的表达显著低于细胞质中的表达(P < 0.05)。在该细胞中转染pcDNA3-TRAF4-DM-TRAF质粒后,其细胞核内TRAF4表达显著升高(P < 0.05), p70S6K及S6蛋白磷酸化水平显著增加( P < 0.05,P < 0.01),S期细胞比例显著增加 (P < 0.01),并且细胞增殖能力显著上调( P < 0.01)。结论 上调TRAF4在乳腺癌细胞MDA-MB-231细胞核中的表达,可能通过促进p70S6K及S6蛋白磷酸化而促进该细胞的增殖。

关键词: 乳腺癌, TRAF4, 细胞增殖

Abstract: Objective To investigate whether overexpression of TRAF4 in human breast cancer may impact its cell proliferation. Methods This study had two groups, MDA-MB-231 transfected with pcDNA3-TRAF4-DM-TRAF or pcDNA3. We detected the expression and localization of TRAF4 with immunofluorescence and western blot. We detected the expression of the phosphorylation level of p70S6K and S6 with westernblot. Flow cytometry detected the cell cycle. MTT Assay detected the cell reproductive capacity. Results TRAF4 localized in both cytoplasm and nuclei in MDA-MB-231. Nuclear expression of TRAF4 in nuclei was lower than that in cytoplasm (P <0.05). After pcDNA3-TRAF4-DM-TRAF was transfected into the cells, the expression of TRAF4 in nuclei was increased (P <0.05). The phosphorylation level of p70S6K and S6 significantly increaseced ( P <0.05,P < 0.01). More S phase cells were detected (P < 0.01) by FCM . The cell proliferation was promoted by MTT (P < 0.01). Conclusion The expression of TRAF4 in nuclei may play an important in the cell proliferatuion by promoting p70S6K and S6 activation.

Key words: Breast cancer, TRAF4, Cell proliferation

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