[急性髓系白血病患者中DKK-3和WIF-1的甲基化状态及mRNA表达]

[Methylation Status and mRNA Expression of DKK-3 and WIF-1 in Acute Myeloid Leukemia Patients].

作者信息

Xu Meng-Meng, Yang Yan-Li, Geng Ying-Hua, Li Jun, Ma Yue, Zhang Feng, Zhu Fang-Bing

机构信息

Department of Hematology, The First Affiliated Hospital of Bengbu Medical College, Bengbu 233000, Anhui Province, China.

Department of Hematology, The First Affiliated Hospital of Bengbu Medical College, Bengbu 233000, Anhui Province, China. E-mail:

出版信息

Zhongguo Shi Yan Xue Ye Xue Za Zhi. 2017 Oct;25(5):1314-1320. doi: 10.7534/j.issn.1009-2137.2017.05.007.

DOI:10.7534/j.issn.1009-2137.2017.05.007

PMID:29070101

Abstract

OBJECTIVE

To analyze the promoter methylation status, mRNA expression and clinical significance of DKK-3 and WIF-1 genes in patients with acute myeloid leukemia(AML).

METHODS

Methylation specific polymerase chain reaction (MS-PCR) mothod was carried out to detect DKK-3 and WIF-1 gene promoter methylation status in bone marrow specimen from 56 patients with AML and 20 patients with iron deficiency anaemia(IDA) as control; then the real-time quantitative reverse transcription polymerase chain reaction (RT-PCR) was used to detect mRNA expression of DKK-3, WIF-1 gene and β -catenin in the above-mentioned specinens, and their relationship with the clinical features and survival time was analyzed.

RESULTS

The promoter methylation rate of DKK-3 and WIF-1 gene in AML patients were significantly higher than that in control group(χ=15.330,P<0.001; χ=17.371,P<0.001). There was no relationship between DKK-3 and WIF-1 gene promoter methylation rate and AML patient's sex, age, clinical typing. The relative expression of DKK-3 and WIF-1 gene mRNA in AML group were 0.840±0.320 and 0.792±0.313, which were lower than those in control group (1.134±0.392 and 1.047±0.334) respectively, the difference was statistically significant (t=3.415,P=0.000; t=3.070, P=0.003). The relative expression of β-catenin mRNA in AML bone marrow specimens in AML group was 0.756±0.304, which was higher than that in control group(0.342±0.105), the difference was statistically significant (t=5.943, P=0.001). The expression of DKK-3 and WIF-1 gene mRNA negatively correlated with β-catenin mRNA(r=-0.543; r=-0.562). Kaplan Meier survival curve analysis showed that overall survival time in AML patients with DKK-3 gene methylation was shorter than that in the AML patients with DKK-3 gene unmethylation(χ=3.957, P=0.042). Futhermore, the orerall survival time in AML patients with WIF-1 gene methylation was also shorter than that in AML patients with WIF-1 gene unmethylation (χ=4.520, P=0.029).

CONCLUSION

Wnt/β-catenin signaling pathway is abnormally activated in AML patients, the DKK-3 and WIF-1 gene promoter methylation may be involved in Wnt pathways activation and the pathogenesis of AML.

摘要

目的

分析急性髓系白血病（AML）患者中DKK-3和WIF-1基因的启动子甲基化状态、mRNA表达及临床意义。

方法

采用甲基化特异性聚合酶链反应（MS-PCR）方法检测56例AML患者骨髓标本及20例缺铁性贫血（IDA）患者骨髓标本作为对照中DKK-3和WIF-1基因启动子甲基化状态；然后采用实时定量逆转录聚合酶链反应（RT-PCR）检测上述标本中DKK-3、WIF-1基因及β-连环蛋白的mRNA表达，并分析其与临床特征及生存时间的关系。

结果

AML患者中DKK-3和WIF-1基因启动子甲基化率显著高于对照组（χ=15.330，P<0.001；χ=17.371，P<0.001）。DKK-3和WIF-1基因启动子甲基化率与AML患者的性别、年龄、临床分型无关。AML组中DKK-3和WIF-1基因mRNA的相对表达分别为0.840±0.320和0.792±0.313，低于对照组（1.134±0.392和1.047±0.334），差异有统计学意义（t=3.415，P=0.000；t=3.070，P=0.003）。AML组AML骨髓标本中β-连环蛋白mRNA的相对表达为0.756±0.304，高于对照组（0.342±0.105），差异有统计学意义（t=5.943，P=0.001）。DKK-3和WIF-1基因mRNA的表达与β-连环蛋白mRNA呈负相关（r=-0.543；r=-0.562）。Kaplan Meier生存曲线分析显示，DKK-3基因甲基化的AML患者总生存时间短于DKK-3基因未甲基化的AML患者（χ=3.957，P=0.042）。此外，WIF-1基因甲基化的AML患者总生存时间也短于WIF-1基因未甲基化的AML患者（χ=4.520，P=0.029）。