摘要
【目的】探讨组蛋白去乙酰化酶抑制剂(HDACIs)抑制神经胶质瘤细胞增殖的机制。【方法】体外培养U251和H4细胞,用组蛋白去乙酰化酶抑制剂:包括帕比司他(LBH589)和M344、伏立诺他(SAHA)处理后,MTT比色法检测细胞存活率,流式细胞术检测细胞周期进程改变,Western blotting和RT-qPCR法分别检测minichromo⁃some maintenance(MCM)蛋白家族成员MCM2、MCM3、MCM4、MCM5、MCM6和MCM7的mRNA和蛋白表达水平。用组蛋白去乙酰化酶抑制剂:包括帕比司他和曲古霉素(TSA)处理U251细胞后,BrdU掺入实验检测细胞增殖和DNA复制能力。用MCM抑制剂环丙沙星(CPX)处理U251和H4细胞,MTT法检测细胞存活率,流式细胞术检测细胞周期进程改变。【结果】与对照组相比,HDACIs降低了U251和H4活细胞数量(P<0.05);HDACIs组BrdU的掺入率降低(P<0.05);HDACIs减少细胞周期S期的比率(P<0.05);Western blotting和qPCR结果显示,HDACIs降低MCM2-7 mRNA和蛋白表达水平(P<0.05);与对照组相比,CPX降低了U251和H4活细胞数量(P<0.05),减少细胞周期S期的比率(P<0.05)。【结论】HDACIs通过下调MCM2-7表达抑制神经胶质瘤细胞增殖。
【objective】To investigate the mechanism of histone deacetylase inhibitors(HDACIs)in suppressing glio⁃ma cell proliferation.【Methods】Glioma cell lines U251 and H4 were cultured in vitro and treated with HDACIs,including LBH589,M344 and SAHA,MTT assay,flow cytometry,RT-qPCR assay and western blotting were perfomed to deter⁃mine the cell viability,cell cycle progression,mRNA and protein expression of minichromosome maintenance protein fam⁃ily(MCM2-7),respectively.BrdU assay was performed to detect the DNA replication in the U251 cells after they were treated with 0.5µmol/L LBH589 and 0.5µmol/L TSA.MTT assay and flow cytometry were performed to determine the via⁃bility and cell cycle progression of U251 and H4 cells respectively after they were treated with ciprofloxacin(CPX).【Results】Compared with those in control group,in HDACIs group,the cell viability was significantly decreased;the viability of cells treated with 0.5µmol/L LBH589 for 12,16,24 h was significantly lower and the longer the LBH589 treatment,the lower the viability(P<0.05).The percentage of U251 and H4 cells in S-phase was significantly lower(P<0.05).The mRNA and protein expression of MCM2-7 was significantly decreased(P<0.05).BrdU incorporation rate was lower.The cell viability and S-phase cell percentage of U251 and H4 cells treated with 0.5 mmol/L LBH589 in CPX group were signif⁃icantly decreased(both P<0.05).【Conclusion】HDACIs suppress glioma cell proliferation via downregulating MCM2-7 ex⁃pression.
作者
李慧锋
袁忠民
吴森斌
马莹
赵凡一
何伟文
梁建峰
伍健伟
LI Hui-feng;YUAN Zhong-min;WU Sen-bin;MA Ying;ZHAO Fan-yi;HE Wei-wen;LIANG Jian-feng;WU Jian-wei(Institute of Neuroscience,The Second Affiliated Hospital of Guangzhou Medical University,Guangzhou 510260,Chi-na;Neurosurgery Department,The Second Affiliated Hospital of Guangzhou Medical University,Panyu District,Guangzhou 511400,China;Guangdong-Hong Kong-Macao Greater Bay Area Center for Brain Science and Brain In-spired Intelligence,Guangzhou 510515,China)
出处
《中山大学学报（医学科学版）》
CAS
CSCD
北大核心
2022年第4期530-538,共9页
Journal of Sun Yat-Sen University:Medical Sciences
基金
广州市科学技术局基金(202002020088)
广东省重点领域研发计划(2018B030340001)
广东省自然科学基金(2019A1515010473)
国家科技部纵向子课题(2016YFC1301703)。
