MS-275对原代培养的胶质瘤细胞及其干细胞生物学行为和耐药性的影响

MS-275 regulates biological behavior and drug resistance of primary cultured glioma cells and glioma stem cells through PI3K/Akt signaling pathway

目的探讨组蛋白去乙酰化酶(HDAC)抑制剂MS-275对原代培养的胶质瘤细胞及其干细胞生物学行为和耐药性的影响。方法取术中切除的脑胶质瘤瘤体深部无囊性变、无坏死肿瘤组织标本,分离获得胶质瘤细胞(GC)和胶质瘤干细胞(CSC),细胞分为4组:GC组,CSC组,GC+MS-275组,CSC+MS-275组。GC组和CSC组不做药物处理;GC+MS-275组和CSC+MS-275组给予MS-275处理24 h(终浓度为8 mmol/L)。检测各组细胞HDAC水平;MMT法检测细胞增殖能力;免疫印迹法检测耐药信号通路相关蛋白(ABCG2、MPR1、MPR 4、Bax、Bcl-2、PI3K、p-PI3K)的表达;流式细胞仪检测细胞周期和细胞凋亡。结果MS-275显著降低GC和GSC中HDAC水平(P<0.05),显著抑制GC和GSC增殖能力(P<0.05),显著增加G0/G1期细胞比例和细胞凋亡率(P<0.05),显著增高GC和GSC中MPR4、ABCG2、MPR1、Bax、p-PI3K表达水平(P<0.05),而显著降低Bcl2表达水平(P<0.05)。与GC+MS-275组比较,GSC+MS-275组变化更明显(P<0.05)。结论MS-275促进GC和CSC凋亡,而且对CSC作用更强,其机制可能与PI3K/Akt信号通路有关。

Objective To explore the effect of histone deacetylases(HDAC)inhibitor MS-275 on the biological behavior and drug resistance of primary cultured glioma cells(GC)and glioma stem cells(GSC).Methods The specimens of gloma tissues removed during the operation were separated to obtain GC and CSC.The cells were divided into 4 groups:GC group,CSC group,GC+MS-275 group,and CSC+MS-275 group.The cells in the GC and CSC groups were not treated with MS-275.The cells in the GC+MS-275 and CSC+MS-275 groups were treated with MS-275 for 24 h with a final concentration of 8 mmol/L.The HDAC levels of cells in each group were detected.MMT method was used to detect the cell proliferation ability.Western blot method was used to detect the expression of drug resistance signal pathway related proteins including ABCG2,MPR1,MPR 4,Bax,Bcl-2,PI3K,and p-PI3K.Flow cytometry was used detect the cell cycle and apoptosis.Results MS-275 significantly reduced the HDAC levels in GC and GSC(P<0.05),significantly inhibited the proliferation ability of GC and GSC(P<0.05),significantly increased the G0/G1 phase cell ratio and apoptosis rate of GC and GSC(P<0.05),significantly increased the expression levels of MPR4,ABCG2,MPR1,Bax,and p-PI3K in GC and GSC(P<0.05),and significantly reduced the expression levels of Bcl2 in GC and GSC(P<0.05).Compared with the cells in GC+MS-275 group,the levels of HDAC and Bcl2 significantly reduced,the G0/G1 phase cell ratio and apoptosis rate,and the expression levels of MPR4,ABCG2,MPR1,Bax,and p-PI3K significantly increased in the cells of GSC+MS-275 group(P<0.05).Conclusions MS-275 may promote the apoptosis of primary cultured GC and CSC and the promotion of CSC is stronger than that of GC,which may be related to the PI3K/Akt signaling pathway.