Shao Li-Li, Fan Juan, Wang Rui, Feng Li-Li, Zhen Chang-Qing, Sui Xiao-Hui, Li Ying
Department of Hematology, Shandong Provincial Hospital Affiliated to Shandong University, Jinan 250021, Shandong Province, China.
Department of Hematology, Shandong Provincial Hospital Affiliated to Shandong University, Jinan 250021, Shandong Province, China. E-mail:
Zhongguo Shi Yan Xue Ye Xue Za Zhi. 2014 Aug;22(4):920-6. doi: 10.7534/j.issn.1009-2137.2014.04.008.
The aim of this study was to detect the mRNA expression of tissue factor pathway inhibitor-2 ( TFPI-2) and its methylation in bone marrow mononuclear cells from acute myeloid leukemia (AML) patients and to explore its significance in AML. Bone marrow mononuclear cells were isolated from newly diagnosed AML patients (n = 33), complete remission AML patients (n = 19), relapsed/refractory AML patients (n = 12) and iron deficiency anemia patients (control group, n = 15). Expression of TFPI-2 mRNA was detected with real-time quantitative PCR (RT-PCR) and the methylation of CpG island in its promoter was detected with methylation-specific PCR (MSP). The results showed that the expression of TFPI-2 mRNA in newly diagnosed AML, complete remission AML and relapsed/refractory AML patients was much lower than that in the controls (P < 0.05). Furthermore, its expression in relapsed/refractory AML patients was lower than that in newly diagnosed AML patients (P = 0.006). Compared with complete remission AML patients, the expression of TFPI-2 mRNA in newly diagnosed AML patients was significantly reduced (P = 0.030). The percentage of TFPI-2 promoter methylation in AML patients was 64.63% (42/64). In newly diagnosed AML group, complete remission AML group and relapsed/refractory AML group,the percentages of TFPI-2 promoter methylation were 66.67% (22/33), 52.63% (10/19) and 83.33% (10/12) (P > 0.05), respectively. The optical density ratio of TFPI-2 mRNA expression was 0.165 (0.005-2.099) in methylated AML patients, and 0.597 (0.011-2.787) in unmethylated AML patients (P < 0.05). Methylation of TFPI-2 gene promoter was not detected in control patients. After 2 courses of chemotherapy, the level of TFPI-2 mRNA was much higher in the CR group than that in the non-CR group (P < 0.05). It is concluded that the down-regulation or silence of TFPI-2 gene potentially results from its promoter methylation, and the expression level of TFPI-2 and the methylation status of its promoter may be used as indicators of risk stratification and evaluation of disease progress.
本研究旨在检测急性髓系白血病（AML）患者骨髓单个核细胞中组织因子途径抑制物-2（TFPI-2）的mRNA表达及其甲基化情况，并探讨其在AML中的意义。从新诊断的AML患者（n = 33）、完全缓解的AML患者（n = 19）、复发/难治性AML患者（n = 12）和缺铁性贫血患者（对照组，n = 15）中分离出骨髓单个核细胞。采用实时定量PCR（RT-PCR）检测TFPI-2 mRNA的表达，采用甲基化特异性PCR（MSP）检测其启动子区CpG岛的甲基化情况。结果显示，新诊断的AML患者、完全缓解的AML患者和复发/难治性AML患者中TFPI-2 mRNA的表达均显著低于对照组（P < 0.05）。此外，复发/难治性AML患者中TFPI-2 mRNA的表达低于新诊断的AML患者（P = 0.006）。与完全缓解的AML患者相比，新诊断的AML患者中TFPI-2 mRNA的表达显著降低（P = 0.030）。AML患者中TFPI-2启动子甲基化的比例为64.63%（42/64）。在新诊断的AML组、完全缓解的AML组和复发/难治性AML组中，TFPI-2启动子甲基化的比例分别为66.67%（22/33）、52.63%（10/19）和83.33%（10/12）（P > 0.05）。甲基化的AML患者中TFPI-2 mRNA表达的光密度比值为0.165（0.005 - 2.099），未甲基化的AML患者中为0.597（0.011 - 2.787）（P < 0.05）。对照组患者未检测到TFPI-2基因启动子的甲基化。化疗2个疗程后，完全缓解（CR）组中TFPI-2 mRNA的水平显著高于未完全缓解（non-CR）组（P < 0.05）。结论是，TFPI-2基因的下调或沉默可能是由于其启动子甲基化所致，TFPI-2的表达水平及其启动子的甲基化状态可作为风险分层和疾病进展评估的指标。
