解剖学报

解剖学报 ›› 2019, Vol. 50 ›› Issue (1): 72-76.doi: 10.16098/j.issn.0529-1356.2019.01.012

• 肿瘤生物学 • 上一篇    下一篇

Survivin shRNA对食管癌细胞c-myc、核因子-κB 基因表达的影响

封敏1 玛依热•吐尔洪2 董娟娟2 木萨•艾尔肯2 李秀梅2*   

  1. 1.新疆医科大学附属肿瘤医院检验科，乌鲁木齐 830011; 2.新疆医科大学基础医学院形态学中心，乌鲁木齐 830011
  • 收稿日期:2018-02-12 修回日期:2018-05-01 出版日期:2019-02-06 发布日期:2019-04-18
  • 通讯作者: 李秀梅 E-mail:dr.lxm@163.com
  • 基金资助:
    miRNA143 靶向 Survivin 调控食管鳞癌细胞凋亡的机制

 Effect of Survivin shRNA on c-myc and nuclear factor-κB expression in esophageal cells

FENG Ming1 Myire•Tuerhong2 DONG Juan-juan2 Msa•Aerken2 LI Xiu-mei 2*   

  1. 1. Clinical Laboratory, the Affiliated Toumour Hospital; 2. Center for Morphology, School of Basic Medicine, Xinjiang Medical University, Urumchi 830011, China
  • Received:2018-02-12 Revised:2018-05-01 Online:2019-02-06 Published:2019-04-18
  • Contact: LI Xiu-mei E-mail:dr.lxm@163.com

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摘要：

目的 探讨Survivin对c-myc、核因子（NF）-κB基因的调控作用。 方法 shRNA干扰沉默食管癌ECA109细胞系中Survivin基因表达，观察c-myc、NF-κB基因的表达变化，流式细胞术检测细胞周期的改变。 结果 1. 食管癌ECA109细胞系中，Survivin shRNA干扰下调Survivin表达后，c-myc mRNA及蛋白的表达水平明显降低；2. Survivin表达后，NF-κB mRNA及NF-κB 总蛋白无明显变化，但其蛋白活化表达水平明显降低, NF-κB 上游调控核因子κB抑制物激酶（IKK）α、IKKβ mRNA表达下调；3. Survivin 表达下调导致G2期延长，G2期细胞比例增加，细胞增殖阻滞。 结论 Survivin在转录及蛋白水平对c-myc具有正向调控作用，Survivin表达下调抑制了IKKα、IKKβ mRNA的表达及NF-κB的磷酸化。

关键词: Survivin shRNA, c-myc, 核因子-κB, 核因子κB抑制物激酶α, 核因子κB抑制物激酶β, 免疫印迹法, 流式细胞术

Abstract:

Objective To study the effect of Survivin shRNA interference on c-myc and NF-κB expression levels in esophageal cells ECA109. Methods Survivin shRNA interference technology was applied to liposomes transfection esophageal ECA109 cells. Half quantitative RT-PCR and Western blotting method were applied to detect genes expression of c-myc and NF-κB gene. Flow cytometry was used to detect the proliferation and apoptosis of cell lines index changes. Results 1. After Survivin shRNA transfected esophageal ECA109 cells, Survivin mRNA and protein expression obviously down-regulated, and c-myc mRNA and protein expression also down-regulated; 2. After Survivin down-regulated, NF-κB mRNA expression had no obvious change, however the phosphorylated proteins level of NF-κB was reduced, whose upstream factor nuclear factor kappa B inhibitor kinase predominate(IKK) alpha, IKK beta mRNA express was cut; 3. Esophageal cell ECA109 apoptosis increased significantly. Cells ratio increased in G2 phase but decreased in S phase. The cell cycle blocked. Conclusion Survivin is effective on adjusting c-myc gene expression on mRNA and protein level. Survivin regulates NF-κB activation by down-regulation of IKK alpha, IKK beta mRNA expression.

Key words: Survivin shRNA, c-myc, Neclear factor-κB, Nuclear factor kappa &beta, inhibitor kinase α、Nuclear factor kappa &beta, inhibitor kinaseβ, Western blotting, Flow cytometry