Chen Ming-Sheng, Zhao Hai-Kang, Cheng Ying-Ying, Yuan Zhi-Hai, Zhang Yue-Lin
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.
Department of Neurosurgery, the Second Affiliated Hospital of Xi'an Medical University, Xi'an 710038, China.
Zhongguo Ying Yong Sheng Li Xue Za Zhi. 2020 Nov;36(6):616-621. doi: 10.12047/j.cjap.5958.2020.129.
To investigate the toxic effects of vitamin C (VC) combined with temozolomide (TMZ) on gliomas and its mechanism. Human glioma cells BMG-1 and SHG44 cells were cultured 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. Compared with the control group, the survival rate of glioma cells in the TMZ group was decreased significantly(＜0.05). Compared with the TMZ group, the survival rate of glioma cells in the VC+TMZ group was decreased significantly(＜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 ＜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 (＜0.05). 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.
探讨维生素C（VC）联合替莫唑胺（TMZ）对胶质瘤的毒性作用及其机制。将人胶质瘤细胞BMG-1和SHG44细胞培养后，具体分为对照组（不加VC和TMZ）、TMZ组（0.2 mmol/L）、VC（0.5 mmol/L）+TMZ（0.2 mmol/L）组和TMZ（0.2 mmol/L）+U0126（10 μmol/L）组，每个实验重复3次。采用MTT法检测细胞存活率；通过流式细胞术和Annexin V-FITC/PI染色检测细胞凋亡；用ROS检测试剂盒检测活性氧（ROS）水平，并用蛋白质免疫印迹法检测凋亡、自噬及ERK通路相关蛋白的表达水平。与对照组相比，TMZ组胶质瘤细胞存活率显著降低（＜0.05）。与TMZ组相比，VC+TMZ组胶质瘤细胞存活率显著降低（＜0.01），细胞凋亡率升高，Bax、Cleaved caspase-3和Cleaved PARP蛋白表达增加，而Bcl-2表达降低。VC+TMZ组ROS水平和自噬率降低，同时LC3-II/LC3-I表达降低，p62表达增加（均＜0.05）。同时，VC联合TMZ降低了BMG-1和SHG44细胞中p-ERK1/2相关蛋白的表达水平，并提高了凋亡率（＜0.05）。VC联合替莫唑胺可增强胶质瘤细胞的毒性。这种作用是通过调节ERK信号通路促进凋亡并抑制替莫唑胺介导的自噬。
