天津医药

天津医药 ›› 2021, Vol. 49 ›› Issue (2): 125-130.doi: 10.11958/20201721

• 细胞与分子生物学 • 上一篇    下一篇

过表达miRNA-495联合替莫唑胺对神经胶质瘤细胞增殖和凋亡的影响 #br#

张园园,朱淑霞,张越华,张燕燕   

  1. 滨州医学院附属医院儿科（邮编256600）
  • 收稿日期:2020-06-18 修回日期:2020-10-31 出版日期:2021-02-15 发布日期:2021-02-02
  • 通讯作者: 朱淑霞 E-mail:zhuxs2003@163.com
  • 基金资助:
    山东省中医药科技发展计划项目

The effect of over-expression of miRNA-495 combined with temozolomide on proliferation and apoptosis of glioma cells #br#

  • Received:2020-06-18 Revised:2020-10-31 Published:2021-02-15 Online:2021-02-02

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摘要： 目的探讨过表达微小RNA（miRNA）-495联合替莫唑胺（TMZ）对神经胶质瘤细胞增殖、凋亡的影响及其
作 用 机 制 。 方法 将 miRNA-495 类 似 物（miRNA-495 mimics）以 及 阴 性 对 照 序 列（miRNA-495 NC）应 用
LipofectamineTM 2000转染至神经胶质瘤U251、A172细胞中，并设空白对照组。采用实时荧光定量PCR（qPCR）检测
转染后miRNA-495、高迁移率族核小体结合域5（HMGN5）mRNA的相对表达量；Western blot法检测HMGN5及基质金
属蛋白酶-9（MMP-9）蛋白表达水平；CCK-8检测神经胶质瘤细胞的增殖能力。miRNA-495 mimics和TMZ单独或联
合作用于U251细胞后，应用CCK-8、流式细胞术检测神经胶质瘤细胞增殖及凋亡情况。 结果在神经胶质瘤U251、
A172 细胞中，与空白对照组和 miRNA-495 NC 组比较，miRNA-495 mimics 组 miR-495 相对表达量升高，HMGN5
mRNA相对表达量及HMGN5、MMP-9蛋白表达量均降低；且细胞增殖能力明显减弱（P＜0.05）。在神经胶质瘤U251
细胞中，与TMZ组和miRNA-495 mimics组比较，TMZ+miRNA-495 mimics组细胞增殖能力明显被抑制，细胞凋亡明
显增加。 结论过表达miRNA-495与化疗药物TMZ联合应用可协同抑制神经胶质瘤细胞增殖、促进细胞凋亡；其机
制可能是通过抑制HMGN5/MMP-9信号通路实现的。

关键词: 神经胶质瘤, HMGN蛋白质类, 基质金属蛋白酶9, 细胞增殖, 细胞凋亡, miRNA-495, HMGN5, 替莫唑胺

Abstract: Objective To explore the effect of over-expression of miRNA-495 combined with temozolomide (TMZ) on
the proliferation and apoptosis in the glioma cells and the related mechanisms. Methods MiRNA-495 mimics and negative
control sequences (miRNA-495 NC) were transfected into U251 cells and A172 cells by LipofectamineTM 2000 in glioma.
The relative expression levels of miRNA-495 and HMGN5 mRNA were detected by quantitative real-time PCR (qPCR). The
protein expression levels of HMGN5 and MMP-9 were detected by Western blot assay. The effects of transfection on the
proliferation of U251 and A172 cells were detected by CCK-8. After the action of miRNA-495 mimics and TMZ alone or in
combination on U251 cells, the apoptosis rate and proliferation inhibition rate of U251 cells were detected by flow cytometry
and CCK-8 assay. Results In the glioma U251 and A172 cells, the relative expression of miRNA-495 was increased in the
miRNA-495 mimics group compared with that of the blank control group and the miRNA-495 NC group. The relative
expressions of HMGN5 mRNA, HMGN5 and MMP-9 protein were decreased in the miRNA-495 mimics group compared with those of the blank control group and the miRNA-495 NC group. The ability of cell proliferation was significantly
decreased (P＜0.05). Compared with the TMZ group and the miRNA-495-mimics group, the cell proliferation in U251 cells
was significantly inhibited, and the apoptosis was significantly decreased in TMZ+miRNA-495 mimics group. Conclusion
The over expression of miRNA-495 and in combination with TMZ can co-inhibit the proliferation and promote apoptosis of
glioma cells, which may be related with the inhibition of HMGN5/MMP-9 signaling pathway.

Key words: glioma, HMGN proteins, matrix metalloproteinase 9, cell proliferation, apoptosis, miRNA-495, HMGN5,
temozolomide

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