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.
ABSTRACT
Objective To assess the safety and efficacy of stent-assisted coil embolization for acutely ruptured wide-necked intracranial aneurysms. Methods We retrospectively reviewed 192 wide-necked intracranial aneurysms in 178 patients. The efficacy and peri-procedure complications of stent-assisted embolization were compared between rup?ture aneurysms and unrupture aneurysms. Results Stent was successfully implanted in 78 rupture aneurysms and 114 un?rupture aneurysms. There was statistically significant difference between rupture aneurysms and unrupture aneurysms groups in rate of poor prognosis on discharge ( 23.1%vs. 5%,χ2=12.726, P0.05)nor in the rate of mortality and permanent disability (8.9%vs. 6.1%,χ2=0.475, P>0.05). Angiograms at 14.7 months of follow-up did not reveal any significant difference between rupture aneu?rysms and unrupture aneurysms groups in aneurysm complete occlusion (74.1%vs. 70.6%,χ2=0.197,P>0.05), recana?lization (10.3%vs. 9.4%,χ2=0.034,P>0.05)and in-stent stenosis (3.4%vs. 4.7%,χ2=0.136,P>0.05). Conclusion Stent-assisted coil embolization for acutely rupture wide-necked intracranial aneurysms can prevent recurrence effective?ly and can achieve high complete occlusion rate in long term follow-up. However, its procedure related complications and mortality is higher in rupture aneurysms than in unrupture aneurysms, which indicates that a caution is needed to conduct stent-assisted coil embolization in rupture aneurysms.
ABSTRACT
Objective To investigate the roles ofc-Jun/Fra1 dimer in viability ofdopaminergic neurons.Methods (1) Dopaminergic neuron-like cell line SH-SY5Y was cultured in vitro.Cells were lysed for immunoprecipitation to determine whether c-Jun dimerized with Fra1.(2) Cells received treatments were divided into four groups:Jun N-terminal kinase (JNK) inhibitor SP600125 treatment group,mitogen-activated kinase (MEK) 1/2 inhibitor U0126 treatment group,SP600125+U0126 treatment group and control group (treated with dimethyl sulfoxide [DMSO]).Western blotting was performed to detect the c-Jun and Fral expression levels and MTT assay was used to observe the cell viability.(3) Cells infected with adenovirus (Ad) carrying c-Jun or Fra1 genes were divided into three groups:Ad-c-Jun group,Ad-Fra1 group and Ad-c-Jun+Ad-Fra1 group,and cells transfected with GFP were used as control group; immunofluorescence and MTT assay were performed to determine the infectious rate and the cell viability,respectively.Results (1) Co-immunoprecipitation showed that the c-Jun dimerized with Fral in SH-SY5Y cell line.(2) Western blotting indicated that inhibition of JNK by SP600125 reduced c-Jun expression and MEK1/2 by U0126 inhibited Fra1 expression; MTT assay indicated that there were significant differences in the cellular viability among the four groups (F=16.647,P=0.000),the cellular viability in control group obviously differed from that in SP600125 treatment group,U0126 treatment group or SP600125+U0126 treatment group (P＜0.05).(3) Immunofluorescence showed that most of cells infected by adenovirus expressed c-Jun or Fra1.MTT assay indicated that there were significant differences in the cellular viability among the four groups (F=14.543,P=0.000),and there were significant differences in their viabilities between control group and Ad-Fra1 group,and between Ad-Fra1 group and Ad-Fra1+Ad-c-Jun group (P＜0.05).Conclusion The c-Jun and Fra1forming a dimer promotes the viability of dopaminergic neuron-like cells.