蛋白酶体抑制剂Lactacystin对胶质瘤C6细胞的增殖抑制作用及其机制

J4 ›› 2010, Vol. 36 ›› Issue (2): 276-280.

蛋白酶体抑制剂Lactacystin对胶质瘤C6细胞的增殖抑制作用及其机制

王海峰¹, 钟加滕², 王伟伟², 葛鹏飞¹, 李文臣¹, 罗毅男¹

1.吉林大学第一医院神经外科| 吉林 |长春 130021； |2.吉林大学基础医学院病理生理学教研室| 吉林长春 130021

收稿日期:2009-12-09 出版日期:2010-03-28 发布日期:2010-03-28
通讯作者: 罗毅男（Tel:0431-88782466,E-mail:luo.yinan@yahoo.com.cn） E-mail:luo.yinan@yahoo.com.cn
作者简介:王海峰（1974-）|男|吉林省松原市人|主治医师|在读医学博士|主要从事胶质瘤综合治疗的研究。
基金资助:
中国博士后基金资助课题（20080440422）；吉林省科技厅科研基金资助课题（200705460）

Inhibitory effect of proteasome inhibitor Lactacystin on proliferation of glioma C6 cells and its mechanism

WANG Hai-Feng¹, ZHONG Jia-Teng², WANG Wei-Wei², GE Peng-Fei¹, LI Wen-Chen¹, LUO Yi-Nan¹

1.Department of Neurosugery,First Hospital,Jilin University,Changchun 130021,China;2. Department of Pathophysiology,School of Basic Medical Sciences,Jilin University,Changchun 130021,China

Received:2009-12-09 Online:2010-03-28 Published:2010-03-28

摘要/Abstract

摘要：

目的：通过观察蛋白酶体抑制剂Lactacystin（LAC）对C6细胞增殖率的影响，探讨LAC引起胶质瘤细胞增殖率发生改变的机制，为临床上治疗胶质瘤提供理论依据。方法：体外培养大鼠胶质瘤C6细胞，选择处于对数生长期的细胞，实验分为对照组、LAC 2.5 μmol/L组、LAC 5.0 μmol/L组及LAC 10.0 μmol/L组，MTT法检测各组C6细胞增殖率，流式细胞术检测细胞凋亡和线粒体膜电势，RT-PCR方法检测各组细胞中Bax和bcl-2 mRNA表达，Western blotting方法检测各组细胞中Bax、bcl-2和P65 蛋白表达。结果：与对照组比较，LAC 5.0 μmol/L组和LAC 10.0 μmol/L组细胞增殖率、线粒体膜电势和P65蛋白表达水平降低（P<0.05），而细胞凋亡率、Bax/bcl-2 mRNA和蛋白的比值增加（P<0.05）。结论：LAC通过诱导细胞凋亡抑制C6细胞的增殖率，其引起凋亡的方式可能是通过线粒体途径，此外在这个过程中NF-κB信号通路也可能影响细胞的增殖率。

关键词: Lactacystin；细胞凋亡；线粒体；细胞增殖

Abstract:

Abstract:Objective To study the mechanism of the changes of proliferation rate induced by the proteasome inhibitor Lactacystin(LAC) through observing the effect of LAC on the proliferation rate of glioma C6 cells, and provide a theoretical basis for clinical treatment of glioma. Methods The cultured rat glioma C6 cells at the logarithmic growth phase were divided into control group,LAC 2.5 μmol/L group,LAC 5.0 μmol/L group and LAC 10.0 μmol/L Group.MTT assay was used to detect the proliferation rate of C6 cells,FCM was used to detect the death and mitochondrial membrane potential of C6 cells and RT-PCR was performed to examine the Bax and bcl-2 mRNA expressions,Western blotting was used to determine the Bax,bcl-2 and P65 protein expressions.Results Compared with control group,the proliferation rates of C6 cells,mitochondrial membrane potential and the level of P65 protein expression in LAC 5.0 and LAC 10.0 μmol/L groups were reduced (P<0.05),while the apoptotic rate,Bax/bcl-2 mRNA and protein ratio were increased (P<0.05).Conclusion LAC can inhibit the proliferation rate of C6 cells by inclucing the apoptosis of C6 cells,which may be related to the mitochondrial pathway,in addition,NF-κB signaling pathway may also affect the proliferation rate of the C6 cells in this process.

Key words: Lactacystin;apoptosis;mitochondria;cell proliferation

中图分类号:

R739.41

王海峰, 钟加滕, 王伟伟, 葛鹏飞, 李文臣, 罗毅男. 蛋白酶体抑制剂Lactacystin对胶质瘤C6细胞的增殖抑制作用及其机制[J]. J4, 2010, 36(2): 276-280.

WANG Hai-Feng, ZHONG Jia-Teng, WANG Wei-Wei, GE Peng-Fei, LI Wen-Chen, LUO Yi-Nan. Inhibitory effect of proteasome inhibitor Lactacystin on proliferation of glioma C6 cells and its mechanism[J]. J4, 2010, 36(2): 276-280.