急性髓系白血病中医证型与ID4基因启动子区甲基化相关性研究

Study on the Correlation between Chinese Medical Syndrome Types and ID4 Gene Promoter Methylation in Human Acute Myeloid Leukemia

目的研究急性髓系白血病(acute myeloid leukemia,AML)患者骨髓细胞中ID4基因启动子区甲基化状态与中医证型的关系,探讨ID4基因甲基化与AML发生、发展的关系。方法选取35例2010年2-12月山东中医药大学附属医院血液科住院或门诊AML患者作为试验组,另选取10名山东中医药大学附属医院健康体检中心体检健康者作为对照组,采用甲基化特异性聚合酶链反应(methylation-specific polymerasechain reaction,MS-PCR)检测两组ID4基因启动子区甲基化状态并进行组间及中医证型间比较,分析中医证型与基因启动子区甲基化相关性。结果 AML患者ID4基因启动子区甲基化为27例(77.1%),与对照组比较,差异有统计学意义(P<0.01)。各证型患者ID4基因启动子区甲基化阳性率由低到高依次为气阴两虚证<瘀血痰结证<毒热炽盛证,差异均有统计学意义(P<0.05)。ID4基因启动子区甲基化阳性患者外周血白细胞数、骨髓原始细胞比例高于阴性患者,差异均有统计学意义(P<0.05)。结论毒热炽盛证、瘀血痰结证患者更易出现ID4基因甲基化,ID4基因启动子区甲基化阳性表达从分子水平验证了AML初期中医邪实内存的本质,也一定程度上反映了AML恶性程度。

Objective To investigate the correlation between Chinese medical syndrome type（CMST） and the ID4 gene promoter methylation state in the bone marrow cells of acute myeloid leukemia（AML） patients,and to discuss the correlation between ID4 gene methylation and the occurrence,development of AML.MethodsThirty-five inpatients or outpatients from the Department of Hematology,the Affiliated Hospital of Shandong University of Traditional Chinese Medicine were recruited as the test group,while 10 healthy volunteers from the health medical center of the Affiliated Hospital,Shandong University of Traditional Chinese Medicine were recruited as the control group.The ID4 gene promoter methylation states were detected using methylation specific polymerase chain reaction（MS-PCR） in the two groups.Inter-group comparison was performed and CMSTs were compared to analyze the correlation between CMSTs and the gene promoter methylation.Results Twenty-seven AML patients（77.1%） had methylation of ID4 gene,showing statistical difference when compared with the control group（P0.01）.The ID4 methylation positive rate of ID4 gene promoter methylation was sequenced from low to high as qi and yin deficiency syndromeinter-accumulation of blood stasis and phlegm syndrometoxic heat inflaming syndrome,showing statistical difference when compared with the control group（P0.05）.The peripheral white blood cells and the bone marrow blast cells were higher in ID4 methylation positive patients than in the ID4 methylation negative control patients with statistical difference（P0.05）.Conclusions Patients of inter-accumulation of blood stasis and phlegm syndrome and toxic heat inflaming syndrome were more likely to have ID4 gene methylation.The ID4 methylation positive expression has verified the essence of evil domination in the early AML at the molecular level.It can also reflect the degree of malignancy of AML to some extent.