摘要
目的探讨地西他滨(DCA)和丙戊酸(VPA)联用对白血病细胞株HL-60的作用及对human peri-od3基因(hPer3)的表达调控的影响。方法 DCA 1.0及4.0μmol·L-1,VPA 2.0 mmol·L-1,VPA 2.0 mmol·L-1+DCA 1.0μmol·L-1,VPA 2.0 mmo.L-1+DCA 4.0μmol·L-1作用HL-60细胞48 h。MTT法检测细胞存活,FITC-AnnexinⅤ/PI检测细胞凋亡,甲基化聚合酶链反应和实时荧光定量PCR检测hPer3基因启动子甲基化状态和mRNA表达,流式细胞术检测CD14表达率。结果 VPA 2.0+DCA 1.0联合用药组生长抑制率为(49.6±5.2)%,VPA 2.0+DCA 4.0联合用药组为(66.3±7.3)%,均高于其相应单药组,差异具有统计学意义(P<0.01)。VPA 2.0+DCA 1.0联合用药组〔早期:(167±3)%,晚期:(32±4)%〕和VPA 2.0+DCA 4.0联合用药组〔早期:(37±5)%,晚期:(36±5)%〕凋亡率均高于其相应单药组,差异具有统计学意义(P<0.01)。VPA 2.0+DCA 1.0联合用药组和VPA 2.0+DCA 4.0联合用药组hPer3启动子甲基化状态均明显低于其相应单药组。VPA2.0+DCA 1.0联合用药组和VPA 2.0+DCA 4.0联合用药组hPer3 mRNA表达分别为1.75±0.33和3.02±0.36,均明显高于其相应单药组,差异具有显著统计学意义(P<0.01)。VPA 2.0+DCA 1.0联合用药组和VPA 2.0+DCA 4.0联合用药组CD14表达率分别为(16.39±1.68)%和(14.82±0.94)%,均明显高于其相应单药组,差异具有显著统计学意义(P<0.01)。结论 DCA联合VPA能显著加强抗白血病效应,作用机制可能与上调hPer3基因的表达相关。
OBJECTIVE To investigate the synergistic effect of decitabine（DCA） and valproic acid（VPA） on human period3（hPer3） gene expression in leukemia HL-60 cells.METHODSCells were treated for 48 h by DCA 1.0,4.0 μmol·L-1,VPA 2.0 mmol·L-1,VPA 2.0 mmol·L-1＋DCA 1.0 μmol·L-1,and VPA 2.0 mmo·L-1＋DCA 4.0 μmol·L-1.Then inhibition rates of cell proliferation were detected by methyl thiazolyl tetrazolium（MTT）.The early apoptosis rate was detected by staining with Annexin Ⅴ and PI.The promoter methylation status of hPer3 gene was detected by MS-PCR.The hPer3 mRNA expressions level was detected by real-time PCR.The CD14 expression was detected by flow cytometry.RESULTS Compared with corresponding VPA or DCA single group,the inhibition rate of VPA 2.0＋DCA 1.0 group and VPA 2.0＋DCA 4.0 was significantly higher（P0.01）,the apoptosis rate of VPA 2.0＋DCA 1.0 group（early period：（16.98±2.85）%,late period：（32.44±4.13）%）and VPA 2.0＋DCA 4.0（early period：（36.87±4.99）%,late period：（35.61±5.22）%）was significantly higher（P0.01）,the promoter methylation status of hPer3 in VPA 2.0＋DCA 1.0 group and VPA 2.0＋DCA 4.0 group was significantly higher（P0.01）,the hPer3 mRNA expression in VPA 2.0＋DCA 1.0 group（1.75±0.33） and VPA 2.0＋DCA 4.0 group（3.02±0.36） was significantly lower（P0.01）,and the CD14 expression in VPA 2.0＋DCA 1.0 group（（14.82±0.94）%） and VPA 2.0＋DCA 4.0 group（（16.39±1.68）%） was significantly lower（P0.01）.CONCLUSION VPA enhances the antileukemia activity of DCA possibly by inducing the expression of hPer3 mRNA.
出处
《中国药理学与毒理学杂志》
CAS
CSCD
北大核心
2012年第1期35-40,共6页
Chinese Journal of Pharmacology and Toxicology
基金
浙江省卫生厅医药科技计划项目(2011KYA169)~~
