维生素C联合替莫唑胺对胶质瘤细胞的毒性作用及其机制*

doi:10.12047/j.cjap.5958.2020.129

中国应用生理学杂志 ›› 2020, Vol. 36 ›› Issue (6): 616-621.doi: 10.12047/j.cjap.5958.2020.129

维生素C联合替莫唑胺对胶质瘤细胞的毒性作用及其机制^*

陈明生¹, 赵海康², 程莹莹², 袁致海², 张越林^3△

1.西安医学院第二附属医院神经外科陕西省脑疾病防治重点实验室, 陕西西安 710038;
2.西安医学院第二附属医院神经外科, 陕西西安 710038;
3.西安医学院陕西省脑疾病防治重点实验室, 陕西西安 710021

收稿日期:2019-10-23 修回日期:2020-11-20 出版日期:2020-11-28 发布日期:2021-03-15
通讯作者: ^△Tel: 029-86131268; E-mail: zhangyuelin68@163.com
基金资助:
^*陕西省脑疾病防治重点实验室(中心)开放课题资助(18NBZD01)

Toxic effects of Vitamin C combined with temozolomide on glioma cells and its mechanism

CHEN Ming-sheng¹, ZHAO Hai-kang², CHENG Ying-ying², YUAN Zhi-hai², ZHANG Yue-lin^3△

1. Department of Neurosurgery, the Second Affiliated Hospital of Xi'an Medical University, Science and Technology Innovation Platform of Shaanxi Key Laboratory of Brain Disorders, Xi'an 710038;
2. Department of Neurosurgery, the Second Affiliated Hospital of Xi'an Medical University, Xi'an 710038, China;
3. Xi'an Medical University, Science and Technology Innovation Platform of Shaanxi Key Laboratory of Brain Disorders, Xi'an 710021, China

Received:2019-10-23 Revised:2020-11-20 Online:2020-11-28 Published:2021-03-15

摘要/Abstract

摘要： 目的:探讨维生素C(VC)联合替莫唑胺(TMZ)对胶质瘤细胞活力的毒性作用及其机制。方法:在体外条件下培养人胶质瘤细胞BMG-1和SHG44细胞,设对照组(不施加VC与TMZ)、TMZ组(0.2 mmol/L)、VC(0.5 mmol/L)+TMZ(0.2 mmol/L)组,TMZ(0.2 mmol/L TMZ)+U0126(10 μmol/L)组,每组实验重复3次。采用MTT实验检测细胞生存率;流式细胞术和Annexin V-FITC/PI染色检测细胞凋亡情况; ROS检测试剂盒检测活性氧簇(ROS)水平, Western blot检测与凋亡、自噬及ERK通路相关蛋白的表达。结果:与对照组比较,TMZ组胶质瘤细胞的存活率显著下降(P＜0.05)。与TMZ组比较,VC+TMZ组胶质细胞瘤细胞的存活率显著下降(P＜0.01),VC+TMZ组中细胞凋亡率显著升高,且Bax、Cleaved caspase-3及Cleaved PARP蛋白表达显著增加,Bcl-2表达显著降低,而ROS水平及细胞自噬率显著降低,LC3-II/LC3-1表达显著降低,p62表达显著增加(P均＜0.05)。同时,联用可降低BMG-1和SHG44细胞中的p-ERK1/2相关蛋白的表达水平,且提高细胞凋亡率(P均＜0.05)。结论:VC联合TMZ能够增强对胶质瘤细胞的毒性,而这一作用是通过ERK信号通路来促进细胞凋亡并抑制替莫唑胺所介导的自噬作用。

关键词: 维生素C, 替莫唑胺, 胶质瘤, 活性氧簇, 细胞毒性

Abstract: Objective: To investigate the toxic effects of vitamin C (VC) combined with temozolomide (TMZ) on gliomas and its mechanism. Methods: Human glioma cells BMG-1 and SHG44 cells were cultured in vitro, specifically divided into control group (without VC and TMZ), TMZ group (0.2 mmol/L), VC (0.5 mmol/L)+TMZ(0.2 mmol/L) group and TMZ(0.2 mmol/L TMZ)+U0126(10 μmol/L)group, each experiment was repeated three times. Cell survival rate was detected by MTT assay; Cell apoptosis was detected by flow cytometry and Annexin V-FITC/PI staining; Reactive oxygen species (ROS) levels were detected by ROS detection kit, and Western blot was used to detect the expression levels of proteins related to apoptosis, autophagy and ERK pathway. Results: Compared with the control group, the survival rate of glioma cells in the TMZ group was decreased significantly(P＜0.05). Compared with the TMZ group, the survival rate of glioma cells in the VC+TMZ group was decreased significantly(P＜0.01), the cell apoptosis rate was increased, and the expressions of Bax, Cleaved caspase-3 and Cleaved PARP protein were increased, while the expression of Bcl-2 was decreased. The ROS level and autophagy rate were decreased, while the expression of LC3-II/LC3-1 was decreased, and the expression of p62 was increased in the VC+TMZ group (all P＜0.05). At the same time, VC combined with TMZ decreased the expression level of p-ERK1/2-related protein in BMG-1 and SHG44 cells, and increased the apoptosis rate (P＜0.05). Conclusion: VC combined with temozolomide can enhance the toxicity of glioma cells. This effect is to promote apoptosis and inhibit temozolomide-mediated autophagy through the regulation of the ERK signaling pathway.

Key words: vitamin C (VC), temozolomide (TMZ), glioma, reactive oxygen species (ROS), cell toxicity

中图分类号:

R739.41

陈明生, 赵海康, 程莹莹, 袁致海, 张越林. 维生素C联合替莫唑胺对胶质瘤细胞的毒性作用及其机制^*[J]. 中国应用生理学杂志, 2020, 36(6): 616-621.

CHEN Ming-sheng, ZHAO Hai-kang, CHENG Ying-ying, YUAN Zhi-hai, ZHANG Yue-lin. Toxic effects of Vitamin C combined with temozolomide on glioma cells and its mechanism[J]. CJAP, 2020, 36(6): 616-621.

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维生素C联合替莫唑胺对胶质瘤细胞的毒性作用及其机制^*

Toxic effects of Vitamin C combined with temozolomide on glioma cells and its mechanism

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