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近年来研究发现ABO血型与许多疾病的发生发展相关,某些肿瘤导致A、B血型物质减少的现象已日益引起关注。本研究探讨ABO基因启动子CpG岛甲基化与白血病的相关性。采用流式细胞仪和激光共聚焦显微镜检测了不同血型的健康人群和各种血液病患者外周血红细胞表面ABH抗原的相对含量,用PCR和MSP-PCR分别检测血液病患者和健康人ABO基因启动子DNA序列和CpG岛甲基化,以及ABO基因启动子-102位点的甲基化。结果发现,白血病患者均出现不同程度的A、B抗原减少;通过对比检测健康人和患者的ABO基因启动子序列,未发现有序列的不同,说明启动子序列高度保守;利用重亚硫酸盐对DNA样本进行修饰后,通过对健康人和患者的ABO基因启动子序列进行扩增和测序,发现健康人和再生障碍性贫血患者在ABO基因启动子的CpG岛区没有甲基化的位点,而急性髓性白血病(AML)、急性淋巴细胞白血病(ALL)、慢性粒细胞白血病(CML)和部分骨髓增生异常综合征(MDS)患者在位置为-102、-101、-100、-99和-97位置的C碱基均有甲基化的现象。结论:甲基化是造成白血病患者AB抗原下降的原因;-102、-101、-100、-99和-97这几个甲基化位点有可能是白血病的特异性表现;针对-102位点检测结果提示-102位点是否甲基化有可能作为白血病鉴别诊断中一个有意义的分子标识物。
In recent years, the study found that ABO blood group is associated with the occurrence and development of many diseases, and some tumors lead to the decrease of A and B blood group substances, which has attracted more and more attention. This study was to investigate the association of ABO gene promoter CpG island methylation with leukemia. Flow cytometry and laser confocal microscopy were used to detect the relative content of ABH antigens on peripheral blood erythrocytes of healthy people of different blood groups and various hematological diseases. The detection of ABO gene promoter in blood sick and healthy people by PCR and MSP-PCR DNA sequence and CpG island methylation, and methylation at the -102 site of the ABO gene promoter. The results showed that patients with leukemia all had different levels of A and B antigens. By comparing the promoter sequences of ABO gene in healthy subjects and patients, there was no sequence difference, indicating that the promoter sequence was highly conserved. Using bisulfite pair DNA samples were modified by healthy and patients ABO gene promoter sequence was amplified and sequenced and found that healthy people and patients with aplastic anemia ABO gene promoter CpG island sites without methylation, Patients with acute myeloid leukemia (AML), acute lymphoblastic leukemia (ALL), chronic myeloid leukemia (CML), and partial myelodysplastic syndrome (MDS) were located at -102, -101, -100, -99 and The C-base at position-97 is methylated. Conclusion: Methylation is the cause of the decrease of AB antigen in leukemia patients. The methylation sites of -102, -101, -100, -99 and -97 may be the specific manifestation of leukemia. Point detection results suggest -102 methylation may be used as a leukemia differential diagnosis of a molecular marker.