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中国应用生理学杂志 ›› 2021, Vol. 37 ›› Issue (6): 601-605.doi: 10.12047/j.cjap.6159.2021.095

• 研究论文 • 上一篇    下一篇

miR-155-3p对HANK1细胞EAF1 mRNA降解速率及恶性增殖的影响*

刘莉1△, 徐凯2, 王禄强3, 徐美丽4   

  1. 1.潍坊市第二人民医院血液科, 山东 潍坊 261041;
    2.阳光融和医院心血管介入科, 山东 潍坊 261000;
    3.阳光融和医院心血管介入科, 山东 潍坊 261000;
    4.潍坊市第二人民医院血液科, 山东 潍坊 261041
  • 收稿日期:2020-09-17 修回日期:2021-03-19 出版日期:2021-11-28 发布日期:2021-11-25
  • 通讯作者: Tel: 15954405896; E-mail: q15954405896@163.com
  • 基金资助:
    *潍坊市奎文区科技发展计划项目(2019第二批第2号)

Effects of miR-155-3p on the degradation rate of EAF1 mRNA and malignant proliferation in HANK1 cells

LIU Li1△, XU Kai2, WANG Lu-qiang3, XU Mei-li4   

  1. 1. Weifang NO. 2 People's Hospital, Weifang 261041;
    2. Sunshine Union Hospital, Weifang 261000;
    3. Sunshine Union Hospital, Weifang 261000;
    4. Weifang NO. 2 People's Hospital, Weifang 261041, China
  • Received:2020-09-17 Revised:2021-03-19 Online:2021-11-28 Published:2021-11-25

摘要: 目的:探究miR-155-3p对人NK/T细胞淋巴瘤细胞HANK1恶性行为的影响及潜在机制。方法:Targetscan数据库预测miR-155-3p的靶基因,培养对数生长期HANK1细胞,将细胞分为空白组、过表达组、对照组及干扰组,利用细胞转染技术依次转入pENTER-puro空白载体、pENTER-miR-155-3p过表达载体、GV248对照载体、GV248-miR-155-3p siRNA干扰载体。同时放线菌素D(ActD)处理各组细胞,实时荧光定量PCR技术检测各组细胞miR-155-3p、EAF1、β-catenin及c-Myc的表达水平,并分析各组细胞ActD处理后EAF1 mRNA降解速率(n=5),Western blot检测细胞EAF1、β-catenin及c-Myc蛋白表达情况(n=3),CCK-8检测细胞恶性增殖能力变化(n=5)。结果:与空白组相比,过表达组细胞miR-155-3p、β-catenin及c-Myc表达水平显著增高,EAF1表达水平降低且EAF1 mRNA半衰期缩短,细胞恶性增殖能力增强(P均<0.05);与对照组相比,干扰组细胞miR-155-3p、β-catenin及c-Myc表达水平显著降低,EAF1表达水平升高且EAF1 mRNA半衰期延长,细胞恶性增殖能力降低(P均< 0.05)。结论:miR-155-3p可促进EAF1 mRNA降解及HANK1细胞恶性增殖能力。

关键词: NK/T细胞淋巴瘤, miR-155-3p, EAF1, 细胞增殖

Abstract: Objective: To investigate the effects and mechanisms of miR-155-3p on the malignant behavior of human NK/T cell lymphoma cell line HANK1. Methods: Targetscan database was used to predict the target gene of miR-155-3p. HANK1 cells in logarithmic growth period were cultured, and the cells were divided into blank group, over-expressed group, control group and interference group, which were transfected with pENTER-puro vector, pENTER-miR-155-3p vector, GV248 control vector and GV248-miR-155-3p siRNA interference vector, respectively. Meanwhile, actinomycin D (ActD) was used to treat each group of cells, and the expressions of miR-155-3p, EAf1, β-catenin and c-Myc in each group were detected by real-time fluorescence quantitative PCR (n=5). The degradation rate of EAF1 mRNA, the expressions of EAF1, β-catenin and c-Myc protein were detected by Western blot (n=3), and the malignant proliferation abilities of cells were detected by CCK-8 (n=5). Results: Compared with the blank group, the expression levels of miR-155-3p, β-catenin and c-Myc in the over-expressed group were significantly higher, the expression level of EAF1 was lower, the half-life of EAF1 mRNA was shortened, and the malignant proliferation ability of the cells was strengthened (P<0.05). Compared with the control group, the expression levels of miR-155-3p, β-catenin and c-Myc in the interference group were significantly lower, and the expression level of EAF1 was increased, the half-life of mRNA was prolonged and the ability of cell proliferation was decreased (P<0.05). Conclusion: miR-155-3p can promote EAF1 mRNA degradation and proliferation in HANK1 cells.

Key words: NK/T cell lymphoma, miR-155-3p, EAF1, cell proliferation

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