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| Inhibition effects of chlorimipramine on human brain U251 glioma cells and its potential mechanism |
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ZHU Xun1, WANG Jing-zhe1, CHEN Pin2, WANG Cun-zu1, XU Hui-zhong3, OUYANG Qi1 |
| （1. School of Medicine, Jiangsu University, Zhenjiang Jiangsu 212013; 2. Department of Neurosurgery, Subei People′s Hospital, Yangzhou Jiangsu 225001; 3. Department of Neurosurgery, Dongtai People′s Hospital, Dongtai Jiangsu 224200, China) |
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Abstract [Abstract]Objective: To explore the possible mechanism of chlorimipramine on human brain glioma U251 cell apoptosis. Methods: Using methyl thiazolyl tetrazolium(MTT) colorimetry with different concentrations of chlorimipramine to test the effects of chlorimipramine on human brain U251 glioma cells; based on the results, selecting two group of different concentrations for the detection of relevant indicators, including death count of trypan blue staining cell, apoptosis staining, the detection of flow cytometry. Additionally, Western blotting and immumofluorescence assay were conducted to examine the expression level of translocator protein(TSPO). DCFHDA probe was performed to examine the level of reactive oxygen species (ROS) and luciferase assay was applied to investigate cell mitochondrial membrane potential. Results: Based on the testing results of MTT, selecting chlorimipramine 57, 28.5 μmol/L, the concentrations of which have statistically significant differences; then putting them into the following tests: trypan blue staining, cell apoptosis staining, and flow cytometry. All the testing results confirmed that chlorimipramine had significantly inhibiting effects on human brain U251 glioma cells, and with a certain degree, the effects were associated with the drug concentration. Additionally, comparing with the control group, Western blotting analysis and immumofluorescence demonstrated that the expression level of TSPO was significantly increased in the groups of chlorimipramine（P<0.05).The level of ROS was significantly higher, but ATP quantity decreased significantly（P<0.05). Conclusion: Chlorimipramine could significantly inhibit human U251 glioma cell growth, promote apoptosis by increasing the expression level of TSPO,ROS and decreasing ATP quantity. Chlorimipramine has good antiglioma effects, and could be used as a kind of candidate drug for the treatment of glioma chemotherapy.
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Received: 12 August 2015
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2016, Vol. 26 
