二甲双胍对胶质母细胞瘤细胞的抑制作用研究

doi:10.19586/j.2095-2341.2022.0062

生物技术进展 ›› 2022, Vol. 12 ›› Issue (6): 937-945.DOI: 10.19586/j.2095-2341.2022.0062

二甲双胍对胶质母细胞瘤细胞的抑制作用研究

孙梦婷(), 赵鹏翔, 仪杨, 王濛, 谢飞, ADZAVON Yao Mawulikplimi, 刘梦昱()

北京工业大学环境与生命学部，北京 100124

收稿日期:2022-04-22 接受日期:2022-05-23 出版日期:2022-11-25 发布日期:2022-11-30
通讯作者: 刘梦昱
作者简介:孙梦婷 E-mail: mtingsun@163.com；
基金资助:
军委后勤保障部开放重点项目(BHJ17L018)

Inhibitory Effect of Metformin Hydrochloride on Glioblastoma Cells

Mengting SUN(), Pengxiang ZHAO, Yang YI, Meng WANG, Fei XIE, Yao Mawulikplimi ADZAVON, Mengyu LIU()

Department of Environment and Life Science，Beijing University of Technology，Beijing 100124，China

Received:2022-04-22 Accepted:2022-05-23 Online:2022-11-25 Published:2022-11-30
Contact: Mengyu LIU

摘要/Abstract

摘要：

为了探究二甲双胍对不同胶质母细胞瘤U87细胞、GL261细胞及C6细胞增殖的影响，选取小鼠GBM细胞GL261细胞系、大鼠GBM细胞C6细胞系及人源GBM细胞U87MG细胞系，使用二甲双胍处理，通过CCK-8法检测细胞增殖活性；细胞实时荧光检测细胞凋亡水平；平板克隆实验检测GBM细胞克隆形成能力；CCK-L法检测胞内ATP水平；Western blot检测Akt及其磷酸化水平。结果显示，与对照组相比，随着作用浓度增加，二甲双胍显著抑制GBM细胞增殖活性，影响细胞形态；与对照组相比，同一作用浓度下，二甲双胍提高了GBM细胞凋亡水平，抑制了GBM细胞克隆形成能力，降低了GBM胞内ATP的产生；二甲双胍处理24 h后，GBM细胞内p-Akt表达显著下调，Akt无明显变化。结果表明，二甲双胍在体外可抑制多种GBM细胞的增殖、克隆，降低胞内ATP水平，其机制可能与Akt磷酸化水平相关，研究结果为进一步探索二甲双胍对胶质母细胞瘤的作用机制提供了体外研究理论基础。

关键词: 胶质母细胞瘤, 二甲双胍, 细胞增殖

Abstract:

The aim of this study was to explore the effect of metformin hydrochloride on the proliferation of different glioblastoma U87 cells， GL261 cells and C6 cells， a mouse GBM cell line GL261， a rat GBM cell line C6 and a human GBM cell line U87MG were selected and treated with metformin hydrochloride. The cell proliferative activity was detected by CCK-8 method； cell apoptosis level was detected by cell real-time fluorescence detection； the clonal formation ability of GBM cells was detected by plate cloning experiment； the intracellular ATP level was detected by CCK-L method； Akt and its phosphorylation were detected by Western blot. Results showed that， compared with the control group， metformin hydrochloride significantly inhibited the proliferative activity of GBM cells and affected cell morphology with the increase of the concentration. Compared with the control group， at the same concentration， metformin hydrochloride increased the apoptotic level of GBM cells， inhibited the clonal formation ability of GBM cells， and reduced the production of ATP in GBM cells. After 24 h of metformin hydrochloride treatment， the p-Akt expression in GBM cells was significantly down-regulated， but there was no significant change in Akt. The results showed that metformin hydrochloride could inhibit the proliferation and clonal formation of a variety of GBM cells and reduce the intracellular ATP level in vitro， which might be related to the phosphorylation of Akt. The results was expected to provide a theoretical basis for further research on the mechanism of metformin hydrochloride against glioblastoma in vitro.

Key words: glioblastoma, metformin hydrochloride, cell multiplication

中图分类号:

R739.91

孙梦婷, 赵鹏翔, 仪杨, 王濛, 谢飞, ADZAVON Yao Mawulikplimi, 刘梦昱. 二甲双胍对胶质母细胞瘤细胞的抑制作用研究[J]. 生物技术进展, 2022, 12(6): 937-945.

Mengting SUN, Pengxiang ZHAO, Yang YI, Meng WANG, Fei XIE, Yao Mawulikplimi ADZAVON, Mengyu LIU. Inhibitory Effect of Metformin Hydrochloride on Glioblastoma Cells[J]. Current Biotechnology, 2022, 12(6): 937-945.

图/表 7

图1 二甲双胍对GBM细胞的增殖活性的影响

Fig. 1 Effect of MET on proliferation activity of GBM cells

图2 二甲双胍对GBM细胞形态的影响(×10)注：MET组与对照组相比，细胞形态发生变化，变圆、皱缩细胞增加，细胞密度降低。

Fig. 2 Effect of MET on GBM cell morphology

图3 二甲双胍对GBM细胞凋亡水平的影响

Fig. 3 Effect of MET on apoptosis of GBM cells

图4 二甲双胍对GBM细胞克隆形成的影响A：克隆形成图；B：GL261细胞；C：C6细胞；D：U87MG细胞；*，**，***表示与对照组相比差异分别在P<0.05、P<0.001和P<0.000 1水平上具有统计学意义。

Fig. 4 Effects of MET on the formation of GBM cell clonies

图5 二甲双胍对GBM克隆细胞团形态的影响

Fig. 5 Effect of MET on cell cluster morphology of GBM clonies

图6 二甲双胍对GBM细胞内ATP水平的影响注：*，**，***表示与对照组相比差异分别在P<0.05、P<0.001和P<0.0001水平上具有统计学意义。

Fig. 6 Effect of MET on intracellular ATP level in GBM cells

图7 二甲双胍对Akt及其磷酸化水平的影响A：GL261细胞；B：C6细胞；C：U87MG细胞；CTRL：对照组；MET：MET处理组；*，***表示与CTRL组相比差异分别在P<0.05和P<0.000 1水平上具有统计学意义。

Fig. 7 Effect of MET on Akt and its phosphorylation level

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二甲双胍对胶质母细胞瘤细胞的抑制作用研究

Inhibitory Effect of Metformin Hydrochloride on Glioblastoma Cells

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