基础医学与临床 ›› 2017, Vol. 37 ›› Issue (8): 1117-1121.

• 研究论文 • 上一篇    下一篇

miR-449a过表达抑制人神经母细胞瘤SH-SY5Y细胞增殖并促进其凋亡

徐晓艳,李小山   

  1. 重庆三峡医药高等专科学校
  • 收稿日期:2016-12-12 修回日期:2017-04-25 出版日期:2017-08-05 发布日期:2017-07-17
  • 通讯作者: 李小山 E-mail:lixiaoshan13@163.com
  • 基金资助:
    重庆市卫计委科技项目

Over-expressed miR-449a inhibits proliferation and induces apoptosis of SH-SY5Y cells in vitro

xiao yanXU,   

  • Received:2016-12-12 Revised:2017-04-25 Online:2017-08-05 Published:2017-07-17

摘要: 目的 探讨miR-449a对人神经母细胞瘤细胞系SH-SY5Y的增殖和凋亡的影响。方法 用LipofectamineTM2000将miR-449a mimics 或 miR-449a control转染至SH-SY5Y细胞,分为blank、miR-449a mimics 和 mi-449a control SH-SY5Y细胞组;实时荧光定量PCR(q-PCR)检测各组细胞中miR-449a表达;CCK8法检测细胞增殖;流式细胞仪检测细胞凋亡和周期,Western blot检测 c-MYC蛋白和Bax/Bcl2蛋白。结果 miR-449a mimics瞬时转染SH-SY5Y细胞后,miR-449a的表达水平明显高于正常对照组(P<0.05);SH-SY5Y细胞增殖能力受到明显抑制(P<0.05);凋亡率明显增加(P<0.05);c-Myc蛋白表达显著降低(P<0.05);细胞促凋亡蛋白Bax表达升高;抗凋亡蛋白Bcl2表达降低(P<0.05)。结论miR-449a可通过c-Myc影响SH-SY5Y细胞的增殖和周期,通过调节Bax/Bcl2影响其凋亡。

关键词: miRNA-449a, SH-SY5Y, 增殖, 凋亡, 周期

Abstract: Objective To explore the effects of miR-449a on the proliferation and apoptosis in human neuroblastoma SH-SY5Y cells. Methods Transferred miR-449a mimics to SH-SY5Y cells by LipofectamineTM2000 as experimental group. Set up blank and negative control group. Cell growth was measured by CCK8 assay, the cell cycle and apoptosis were detected by flow cytometry. Relative expression levels of c-Myc, Bax/ Bcl-2 were detected by Western blot. Results Compare to blank and negative control cells, the proliferation of miR-449a mimics SH-SY5Y cells were decreased(P<0.05). Flow cytometry showed that over-expressed miR-449a resulted in an increase of G0/G1 phase and apoptotic cells. The expression of c-Myc showed significant difference (P<0.05), Expression of pro-apoptotic protein Bax was increased, and that of anti-apoptotic protein Bcl-2 was decreased differentially (P<0.05). Conclusions miR-449a over-expression can inhibit cell proliferation by regulating cell cycles and induce apoptosis mainly through regulating Bcl-2 family proteins expression in SH-SY5Y cells.

Key words: miRNA-449a, SH-SY5Y, proliferation, apoptosis, cell cycle

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