PCR检测降钙素基因甲基化及其在某些恶性血液病中的意义

Detection of the hypermethylation of calcitonin gene by polymerase chain reaction and its implication in some hematological malignancies

目的 :探讨急性非淋巴细胞白血病 (acutenonlymphoblasticleukecia ,ANLL)、骨髓增生异常综合征(myelodysplasticsyndrome ,MDS)和慢性粒细胞白血病 (chronicmyeloidleukemia ,CML)患者降钙素基因 (calcitoningene ,CT基因 )甲基化状况及其意义。方法 :利用限制性内切酶和多聚酶链反应 ( polymerasechainreaction ,PCR)技术。结果 :72 % ( 18/ 2 5 )的ANLL和 6 0 % ( 6 / 10 )的MDS患者有CT基因高度甲基化 ,在CML慢性期CT基因甲基化阳性率为 2 3 .8% ( 5 / 2 1) ,而急变期阳性率为 80 % ( 8/ 10 ) ,两者之间有显著性差异 ,系列稀释法发现本试验敏感性至少达 10 3 水平。结论 :CT基因高度甲基化较广泛地存在于ANLL、MDS和CML急变期 ,是ANLL和MDS患者白血病细胞克隆以及CML进展和急变的一个共同的分子标志 。

Objective:To study the methylation pattern of calcitonin gene (CT gene) in acute nonlymphoblastic leukemia (ANLL ),myelodysplastic syndrome (MDS) and chronic myeloid leukemia (CML) and its implication. Methods:Restriction endonuclease and polymerase chain reaction (PCR) techniques were used. Results:The hypermethylation pattern was found in 72%(18/25) of ANLL and 60%(6/10) of MDS patients.In CML,23.8%(5/21) samples in chronic phase and 80%(8/10) samples in blast crisis were positive ( P< 0. 05) . And the sensitivity of this method was at least 3 logs (1 in 1000). Conclusion:This findings indicated that the hypermethylation of CT gene was a common molecular marker for the tumor cells in ANLL,MDS and the progression and blast crisis of CML.The hypermethylation of CT gene might be associated with the transformation to increase blastic cells in MDS and might have prognostic value in MDS.