GANT61对髓母细胞瘤Daoy细胞增殖、凋亡的影响及作用机制  

Influence of GANT61 on the proliferation and apoptosis of medulloblastoma Daoy cells and its mechanism

作　　者：卢恩敏 卢恩娟 高丽京 裴立静 王庆艳[1] LU Enmin;LU Enjuan;GAO Lijing;PEI Lijing;WANG Qingyan(Department of Pediatrics,People’s Hospital of Fengrun District,Tangshan 063000,Hebei,China;Depatment of Pediatrics,Second People’s Hospital of Fengrun District,Tangshan 063000,Hebei,China)

机构地区：[1]唐山市丰润区人民医院儿科,河北唐山063000 [2]唐山市丰润区第二人民医院儿科,河北唐山063000

出　　处：《癌症进展》2020年第12期1207-1210,共4页Oncology Progress

基　　金：河北省卫生厅科研基金项目(20181399)。

摘　　要：目的探讨GANT61对髓母细胞瘤(MB)Daoy细胞增殖、凋亡的影响及作用机制。方法将人MB Daoy细胞正常培养24 h,然后分为4组:A组,培养液中加入0.1%二甲基亚砜(DMSO);B组,培养液中加入10μmol/L GANT61和0.1%DMSO;C组,培养液中加入20μmol/L GANT61和0.1%DMSO;D组,培养液中加入40μmol/L GANT61和0.1%DMSO。采用噻唑蓝(MTT)法检测细胞增殖,采用实时定量聚合酶链反应(PCR)检测GLI家族锌指蛋白1(GLI1)mRNA、GLI家族锌指蛋白2(GLI2)mRNA的相对表达量,采用蛋白质印迹法(Western blot)检测细胞周期蛋白D1(cyclin D1)、p21反胱天蛋白酶3(caspase 3)的表达水平,采用流式细胞术检测细胞周期。结果培养24 h后,C组和D组细胞的光密度(OD)值均低于A组,差异均有统计学意义(P<0.05);培养48 h后,C组和D组细胞的OD值均低于A组,D组细胞的OD值低于B组和C组,差异均有统计学意义(P<0.05)。C组和D组细胞中GLI1 mRNA和GLI2 mRNA的相对表达量均低于A组,D组细胞中GLI1 mRNA和GLI2 mRNA的相对表达量均低于B组,差异均有统计学意义(P<0.05)。C组和D组细胞中cyclin D1蛋白表达水平均低于A组和B组,p21、caspase 3蛋白表达水平均高于A组和B组,差异均有统计学意义(P<0.05)。B组、C组和D组中G0/G1,期细胞比例均高于A组,G2/M期细胞比例均低于A组,差异均有统计学意义(P<0.05);C组和D组中G0/G1,期细胞比例均高于B组,差异均有统计学意义(P<0.05)。结论GANT61对MB Daoy细胞具有抑制细胞增殖和促进细胞凋亡的作用,可能是通过调节GLI1、GLI2的mRNA水平进而影响cyclin D1、p21及caspase 3蛋白表达来实现。Objective To explore the effect of GANT61 on the proliferation and apoptosis of medulloblastoma(MB)Daoy cells and its mechanism.Method The human MB Daoy cells were cultured for 24 h and then randomly divided into four groups as follows,group A:with 0.1%DMSO in medium solution,group B:with 10μmol/L GANT61 and 0.1%DMSO;group C:with 20μmol/L GANT61 and 0.1%DMSO;group D:with 40μmol/L GANT61 and 0.1%DMSO,respectively.Thiazolyl blue tetrazolium bromide(MTT)assay were utilized to analyze cell proliferation,and real-time polymerase chain reaction(PCR)was used to detect the relative expression level of GLI family zinc finger 1(GLI1)mRNA and GLI family zinc finger 2(GLI2)mRNA in Daoy cells,besides,Western blot was employed to determine the expression of cyclin D1,p21 and caspase 3,and the cell cycle was analyzed using flow cytometry.Result After culturing for 24 h,the optical density(OD)values detected in group C and D were significantly lower than that in group A(P<0.05);at 48 h after culturing,lower OD values were observed in group C and D than in group A,and the OD value in group D was decreased compared to group B and C(P<0.05).Compared with group A,the relative expression of GLI1 mRNA and GLI2 mRNA in group C and D were lower,and it was also lower in group D versus group B(P<0.05).Evidently decreased lower expression of cyclin D1 proteins in group C and D were noted in comparison with group A and B,while p21 and caspase 3 protein expression were higher than those in group A and B(P<0.05).Group B,C and D were with higher proportion of cells in G0/G1 phase,and less G2/M phase cells than in group A(P<0.05);while in group C and D,higher percentage of cells in G0/G1 phase were noted compared to group B(P<0.05).Conclusion GANT61 may inhibit cell proliferation while promote apoptosis for MB Daoy cells,possibly by regulating the levels of GLI1 and GLI2 mRNA,thereby influencing the protein expression of cyclinD1,p21 and caspase 3.

关 键 词：GANT61 Daoy细胞 细胞周期蛋白D1 P21 胱天蛋白酶3 

分 类 号：R739.4[医药卫生—肿瘤]