吉林大学学报(医学版) ›› 2016, Vol. 42 ›› Issue (04): 637-641.doi: 10.13481/j.1671-587x.20160401

• 基础研究 •    下一篇

NPPB对人脑胶质瘤SHG-44细胞凋亡的诱导作用及其机制

田晶1, 齐玲2, 纪朋艳3, 沈楠3, 崔万丽1, 王春艳1   

  1. 1. 吉林医药学院生理学教研室, 吉林 吉林 132013;
    2. 吉林医药学院病理学教研室, 吉林 吉林 132013;
    3. 吉林医药学院实验中心, 吉林 吉林 132013
  • 收稿日期:2015-07-07 出版日期:2016-07-28 发布日期:2016-07-20
  • 通讯作者: 齐玲,副教授,硕士研究生导师(Tel:0432-64560027,E-mail:qiling1718@163.com) E-mail:qiling1718@163.com
  • 作者简介:田晶(1973-),女,吉林省吉林市人,副教授,医学硕士,主要从事脑肿瘤治疗方面的研究。
  • 基金资助:

    国家自然科学基金资助课题(81201671);吉林省教育厅"十二五"科研项目资助课题(2012330,2014369)

Induction effect of NPPB on apoptosis of human glioma SHG-44 cells and its mechanism

TIAN Jing1, QI Ling2, JI Pengyan3, SHEN Nan3, CUI Wanli1, WANG Chunyan1   

  1. 1. Department of Physiology, Jilin Medical College, Jilin 132013, China;
    2. Department of Pathology, Jilin Medical College, Jilin 132013, China;
    3. Experimental Center, Jilin Medical College, Jilin 132013, China
  • Received:2015-07-07 Online:2016-07-28 Published:2016-07-20

摘要:

目的:探讨氯通道阻断剂5-硝基-2-(3-苯丙胺)苯甲酸(NPPB)对人脑胶质瘤SHG-44细胞凋亡的诱导作用,并阐明其可能的机制。方法:体外培养脑胶质瘤SHG-44细胞,分为对照组及50、100和200 μmol·L-1 NPPB组。采用MTT法检测细胞活力,采用流式细胞术检测细胞凋亡率,采用免疫组织化学法和蛋白免疫印记法分别检测SHG-44细胞中凋亡蛋白的表达水平。结果:MTT检测,与对照组比较,作用24和48 h,100和200 μmol·L-1 NPPB组SHG-44细胞活力明显降低(P < 0.01)。流式细胞术检测,100和200 μmol·L-1 NPPB组细胞凋亡率分别为24.64%和41.85%,高于对照组(4.17%)(P < 0.01)。免疫组织化学和蛋白免疫印记法检测,100 μmol·L-1 NPPB组SHG-44细胞中caspase-3和Bax蛋白表达水平升高(P < 0.05或P < 0.01),Bcl-2蛋白表达水平降低(P < 0.05)。结论:NPPB可以通过下调Bcl-2的表达、上调Bax的表达,使caspase-3活化,进而引起SHG-44细胞凋亡。

关键词: 氯通道, 神经胶质瘤, 细胞增殖, 细胞凋亡

Abstract:

Objective:To investigate the induction effect of NPPB,a chloride channel blocker,on the apoptosis of human glioma SHG-44 cells,and to explore its mechanism. Methods: The SHG-44 cells were cultured in vitro and divided into control group and NPPB groups (50,100,200 μmol·L-1). The cell viability was detected by MTT assay. The apoptotic rates were detected by flow cytometry. The expression levels of Bax,Bcl-2 and caspase-3 were detected by immunohistochemical analysis and Western blottingmethod. Results: Compared with control group,the cell viabilities of SHG-44 cells in 100 and 200 μmol·L-1 NPPB groups after treated for 24 and 48 h were decreased significantly(P < 0.01). The results of flow cytometry showed that the apoptotic rates of SHG-44 cells in 100 and 200 μmol·L-1 NPPB groups were 24.64% and 41.85%,and they were higher than that in control group(4.17%)(P < 0. 01).The immunohistochemical analysis and Western blotting results showed that the expression levels of caspase-3 and Bax proteins in SHG-44 cells in 100 μmol·L-1 NPPB group were increased(P < 0.05 or P < 0. 01),and the expression level of Bcl-2 protein was decreased(P < 0.05). Conclusion:NPPB could induce the apoptosis of human glioma SHG-44 cellsby the down-regulation of the expression of Bcl-2 and the up-regulation of the expression of Bax,and the activation of caspase-3.

Key words: chloride channel, glioma, cell proliferation, apoptosis

中图分类号: 

  • R739.41