Jin Zheng, Yu Ying, Jin Ri-Hua, Wang Yu-Bo, Xu Hai-Yang
Department of Neurosurgery, The First Hospital of Jilin University Changchun 130021, Jilin, China.
Am J Transl Res. 2016 Sep 15;8(9):3970-3975. eCollection 2016.
Glioblastoma is a highly malignant cancer of glioma cells. Present study investigates the anti proliferative activity of granatin B on glioma cell by inducing apoptosis. In this study Glioma cell (U87) was used on which anti proliferative activity of granatin B (0, 20, 40 & 80 µM) assessed by 3-(4, 5-dimethylthylthiazol-2-yl)-2,5 diphenyltetrazolium bromide (MTT) assay. Thereafter Apoptosis of glioma cell was assessed by apoptosis detection kit suing flow cytometer, DAPI staining and by estimating the activity of caspase 3 & 9 using caspase 3 & 9 kit. Expression of MMP9 protein was determined through gelatin zymography. Possible mechanism of apoptosis induction was proved by estimating the effect of granatin B with MMP9 agonist on cell proliferation, caspase 3 activity & MMP9 expression on glioblastoma cell. Result of the study suggested that granatin B significantly decreases the cell proliferation of glioma cell compared to 0 µM treated group. It was also observed that treatment with granatin B significantly induces apoptosis and increases the activity of caspase 3 & 9 protein compared to 0 µM treated group. Expression of MMP9 protein was also decreases with granatin B treatment of glima cell. MMP9 agonist significantly reverses the effect of granatin B on cell proliferation, caspase 3 and expression of MMP9 protein in glima cell. Present study concludes the anticancer activity of granatin B on glioblastoma cell by inducing apoptosis.
胶质母细胞瘤是一种由胶质瘤细胞构成的高度恶性肿瘤。本研究通过诱导凋亡来探究格栏亭B对胶质瘤细胞的抗增殖活性。在本研究中，使用了胶质瘤细胞（U87），通过3-(4,5-二甲基噻唑-2-基)-2,5-二苯基四氮唑溴盐（MTT）法评估格栏亭B（0、20、40和80μM）的抗增殖活性。此后，使用凋亡检测试剂盒，通过流式细胞仪、DAPI染色以及使用caspase 3和9试剂盒评估caspase 3和9的活性来评估胶质瘤细胞的凋亡情况。通过明胶酶谱法测定MMP9蛋白的表达。通过评估格栏亭B与MMP9激动剂对胶质母细胞瘤细胞增殖、caspase 3活性和MMP9表达的影响，证明了凋亡诱导的可能机制。研究结果表明，与0μM处理组相比，格栏亭B显著降低了胶质瘤细胞的增殖。还观察到，与0μM处理组相比，格栏亭B处理显著诱导了凋亡并增加了caspase 3和9蛋白的活性。格栏亭B处理胶质瘤细胞后，MMP9蛋白的表达也降低。MMP9激动剂显著逆转了格栏亭B对胶质瘤细胞增殖、caspase 3和MMP9蛋白表达的影响。本研究得出结论，格栏亭B通过诱导凋亡对胶质母细胞瘤细胞具有抗癌活性。
