血液系统疾病中LRP15基因启动子区甲基化状态的研究

LRP15-gene-promoter methylation patterns in hematological diseases

目的:探讨LRP15基因启动子区及第一外显子区的甲基化与血液系统疾病的关系和临床意义。方法:采用甲基化特异性PCR方法检测了9例正常人、73例急性白血病患者、8例慢性髓系白血病患者、2例慢性淋巴细胞白血病、12例血液系统良性病患者骨髓LRP15基因启动子区及第一外显子区的甲基化状况;硫化修饰结合限制性内切酶分析及硫化测序验证了一例血液系统良性病患者LRP15基因启动子区及第一外显子区的甲基化。结果:AL患者LRP15基因甲基化阳性率(71.23%)高于正常供者及慢性白血病(10%),差异有统计学意义(P<0.05)。LRP15基因在非恶性血液病患者中甲基化阳性率(55.56%)高于正常供者,硫化修饰结合限制性内切酶分析及硫化测序进一步证实了上述结论。结论:LRP15基因启动子的甲基化与不成熟的白血病细胞有关,与血液系统良性疾病也有一定关系。

Objective：To explore the methylation status of LRP15 gene in hematological diseases and its role in the tumorigenesis. Methods：Methylation patterns were detected in 9 healthy donors, 73 acute leukemia （AL） patients, 8 chronic myeloid leukemia（CML） patients, 2 chronic lymphocytic leukemia （CLL） patients and 12 patients with benign hematological diseases by methylation specific PCR （MSP）. COBRA （Combined bisulfite restriction analysis） and bisulfite sequence analysis confirmed LRP15 methylation in an adult with benign hematological diseases. Results： The frequency of LRP15 methylation was 71.23% （52/73）in AL patients, 0% （0/9） in healthy donors and 10% （1/10） in CL patients. The diffrence of LRP15 methylation frequency between AL patients and healthy donors was statistically significant （P =0.000）. The difference between AL patients and CL patients was also statistically significant. LRP15 methylation in 55.56% （5/9） adult patients with hematological benign diseases was detected for the first time. COBRA and bisulfite sequence analysis confirmed LRP15 methylation in an adult with benign hematological diseases. Conclusions： LRP15 methylation change is common abnormality in immature leukemia cells. LRP15 may be a putative tumor suppressor gene. It is possible that hypermethylation of LRP15 gene is related to the pathogenesis and development of hematological benign diseases.