Histone deacetylase inhibitors suppress JNK activity-mediated proliferation of glioma cells
10.3760/cma.j.issn.1671-8925.2015.06.002
- VernacularTitle:组蛋白去乙酰化酶抑制剂抑制神经胶质瘤细胞增殖的机制研究
- Author:
Jianwei WU
1
;
Jianfeng LIANG
;
Weiwen HE
;
Zhongmin YUAN
Author Information
1. 510260 广州,广州医科大学附属第二医院神经外科,广州医科大学神经科学研究所(广东省重点实验室,神经遗传与离子通道病省部共建教育部重点实验室)
- Keywords:
Glioma;
Trichostatin A;
JNK;
Proliferation
- From:
Chinese Journal of Neuromedicine
2015;14(6):547-552
- CountryChina
- Language:Chinese
-
Abstract:
Objective To investigate the mechanism of histone deacetylase inhibitors in proliferation ofglioma cells.Methods (1) Glioma cell line U251 was cultured in vitro and treated with Trichostatin A (TSA) at 0.1,0.2,0.5,1.0 or 2.0 μmol/L for 48 h to determine the IC50 for TSA,and then,cells were treated with TSA at the IC50 concentration (0.5 μ mol/L) for 8,16,24,48 h;1 μmol/L M344,0.5 μmol/L LBH589,6 mmol/L NaBu and 6 mmol/L VPA were used to treat the cells for 48 h and same volume of solvent was given to cells for 48 h as control group;MTT assay was performed to determine cell viability.(2) Western blotting was performed to test the Jun N-terminal kinase (JNK)expression and phosphorylated JNK (p-JNK) level in the U251 cells after being treated with 0.5 μmol/L TSA for 8,16 and 24 h.(3) Western blotting and MTT assay were employed to detect the c-Jun expression and phosphorylated c-Jun (p-c-Jun) level,and cell viability in the U251 cells of control group,10 μmol/L SP600125 treatment group (JNK inhibitor) and 1 μmol/L CEP11004 treatment group (MLK3,a direct upstream kinase of JNK).(4) Western blotting and MTT assay were employed to detect the c-Jun and Flag expressions,and cell viability in the U251 cells of pcDNA3.1 transfected group,pcDNA3.1+TSA transfected group,pMKK7-JNK1 transfected group and pMKK7-JNK1+TSA transfected group.Results (1) The viability of cells of 0.1,0.2,0.5,1.0 or 2.0 μmol/L TSA treated group was significantly lower than that in the control group,and the higher the TSA concentration,the lower the viability of cells;the viability of cells of 0.5 μmol/L TSA treated for 16,24,36 and 48 h groups was significantly lower than that in the control group,and the longer the TSA treatment,the lower the viability of cells.(2) The viability of 1 μmol/L M344,0.5 μmol/L LBH589,6 mmol/L NaBu and 6 mmol/L VPA treatment groups was significantly lower than that in the control group (P＜0.05).(3) As compared with those in the control group,the p-JNK expression was significantly decreased in the cells after 0.5 μmol/L TSA treatment for 16 and 24 h,and the p-c-Jun protein expression and the cell viability were significantly decreased in the cells of 10 μmol/L SP600125 and 1 μmol/L CEP11004 treatment groups (P＜0.05).(4) As compared with that in the pcDNA3.1 transfected group,cell viability in the U251 cells ofpcDNA3.1+TSA transfected group,pMKK7-JNK1 transfected group and pMKK7-JNK1+TSA transfected group was significantly increased(P＜0.05).Conclusion Histone deacetylase inhibitors reduce the proliferation of glioma cell U251 through inhibiting JNK activity.
