摘要
目的探讨白血病ABO基因启动子区CpG岛甲基化对ABO基因表达的影响。方法以25例白血病患者及20名健康对照者为研究对象，同时以不同浓度去甲基化药物地西他滨处理白血病细胞株K562、HL．60和Jurkat细胞，采用序列特异性引物PCR（PCR-SSP）法检测ABO基因分型，采用实时荧光定量PCR法检On,4ABOmRNA表达，采用亚硫酸氢盐测序法检测ABO基因启动子区CpG岛甲基化情况。结果①急性淋巴细胞白血病组（10例）和急性髓系白血病组（10例）ABO基因启动子平均甲基化率分别为53．85％和18．22％，明显高于健康对照组（20名，2．33％）和慢性髓性白血病组（5例，2．12％）。②K562细胞的ABO基因型为O1O1，药物作用前后ABO基因型变化不大；HL-60和Jurkat细胞药物作用前基因型无法辨认，药物作用后ABO基因型为O1A1和A1O2。⑧K562细胞ABOmRNA相对表达水平为1275．67：k35．86，明显高于HL-60和Jurkat细胞（O．54±0．07和0．82±0．16）（P值均〈0．05）。@K562、HL-60、Jurkat细胞ABO基因启动子区甲基化率分别为0、58．14％和96．74％；与药物作用前比较，随地西他滨浓度增高，HL-60和Jurkat细胞的甲基化水平下降、ABOmRNA表达水平增高，差异均有统计学意义（P值均〈0．05）。结论ABO基因启动子区甲基化水平与急性白血病ABO基因表达呈负相关，DNA甲基化导致ABO基因沉默是急性白血病ABO抗原表达减弱的重要机制之一。
Objective To investigate the impact of promoter CpG island methylation on ABO mRNA expression in leukemia. Methods 25 cases of leukemia and 20 cases of normal control were studied, and the leukemia cell lines K562, HL-60 and Jurkat were treated with different concentrations of decitabine. PCR-SSP was used to identify ABO genotype, RQ-PCR for ABO mRNA expression and bisulfite sequencing PCR for DNA methylation status. Results （1）The methylation of ABO promoter in acute myeloid leukemia patients （10 cases） and acute lymphoblastic leukemia patients （10 cases） were 53.85% and 18.22% respectively, which were obviously higher than those in control （20 cases, 2.33%） and chronic myeloid leukemia patients （5 cases, 2.12% ）. （2）ABO genotype of K562 was O1O1, which has changed little before and after decitabine treatment. ABO genotype of HL-60 and Jurkat could not been identify before treatment, but showed as O1A1 and A1O2 after treatment. （3）ABO mRNA expression of K562 was 1 275.67±35.86, which was obviously higher than that in HL-60 （0.54±0.07, P〈0.05） and Jurkat （0.82±0.16, P〈0.05）. （4）The methylation of ABO promoter in K562, HL-60 and Jurkat were 0, 58.14%, and 96.74%. As concentration of decitabine increased, the methylation of ABO promoter were decreased and the expressions of ABO mRNA were increased in HL-60 and Jurkat, which had significant differences compared with that before treatment （P〈0.05）. Conclusion The methylation of ABO promoter shows a negative correlation with ABO mRNA expression. DNA methylation was an important aspect of ABO antigens decrease in acute leukemia.
出处
《中华血液学杂志》
CAS
CSCD
北大核心
2016年第9期795-799,共5页
Chinese Journal of Hematology
基金
基金项目：河南省教育厅科学技术研究重点项目（13A320431）
