摘要
目的研究慢性粒细胞白血病(chronic myeloid leukemia,CML)骨髓细胞SFRP2基因启动子甲基化状态,探讨SFRP2基因启动子甲基化在CML发病机制中的作用。方法分别采用RT-PCR、甲基化特异性PCR(methylation-specif-ic PCR,MSP)方法检测CML细胞系K562细胞及骨髓标本SFRP2 mRNA及SFRP2基因启动子CpG岛甲基化状态;并予DNA甲基化转移酶抑制剂5-氮杂-2’-脱氧胞苷(Aza)联合组蛋白去乙酰化酶抑制剂曲古菌素A(TSA)处理K562细胞,观察其SFRP2 mRNA表达情况。结果 SFRP2 mRNA在K562细胞、CML和正常人骨髓标本表达率分别为0、22%(4/18)、100%(8/8)(P<0.05);SFRP2基因启动子甲基化率在K562细胞、CML和正常人骨髓标本分别为100%、66%(25/38)、0(0/13)(P<0.05);伴随K562细胞去甲基化药物处理后未甲基化序列的增加,其SFRP2 mRNA恢复了表达。结论 CML骨髓细胞的SFRP2基因启动子存在高甲基化现象,SFRP2基因启动子甲基化可能是CML发生发展的分子机制之一,提示了其甲基化可能成为CML的新的参考标记物。
Objective To determine the methylation status of secreted frizzled-related protein 2（SFRP2） gene in chronic myeloid leukemia（CML） bone marrow cells,and try to find the role of its promoter methylation during the tumorigenesis of CML.Methods The expression of SFRP2 in CML cell line K562,bone marrow specimens of identified CML patients and health volunteers were detected with semi-quantitative PCR（RT-PCR）.The methylation status of SFRP2 promoter was detected in K562 cells and CML bone marrow specimens by methylation-specific PCR（MSP）.After K562 cells was treated with pharmacological demethylation 5-aza-2＇deoxycytidine（Aza） and trichostatin A（TSA）,the expression of SFRP2 was detected again with RT-PCR.Results SFRP2 was silenced in K562 cells,expressed in 22.0%（4/18） CML bone marrow specimens,and detected in all normal bone marrow samples（100.0%,8/8,P0.05）.The frequencies of SFRP2 promoter methylation were 100%,66%（25/38） and 0（0/13） respectively in K562 cells,CML bone marrow and normal bone marrow specimens（P0.05）.Pharmacological demethylation treatment dramatically induced mRNA expression of SFPR2,and this reactivation was associated with an increase of unmethylation alleles.Conclusion The promoter of SFRP2 gene are hypermethylated in CML,which be one of the molecular mechanisms of CML.Its hypermethylation might serve as a new biomarker for CML.
出处
《第三军医大学学报》
CAS
CSCD
北大核心
2011年第24期2583-2586,共4页
Journal of Third Military Medical University
关键词
慢性粒细胞白血病
SFRP2
甲基化
去甲基化
chronic myeloid leukemia
secreted frizzled-related protein 2
methylation
demethylation
