摘要
目的:本研究通过检测HIF-1α在原代急性髓系白血病(AML)细胞中的表达,探讨其表达水平与临床参数、预后之间的关系。方法:采用CD3单克隆抗体负分选法分选出53例初诊AML(除急性早幼粒细胞白血病APL)患者外周血中的AML细胞,应用实时荧光定量RT-PCR(FQ RT-PCR)检测AML细胞的HIF-1α表达水平,以管家基因β-actin为内参,按2-△△Ct法计算目的基因相对正常供者的表达量,分析其表达水平与患者年龄、性别、白细胞计数、诊断分型、疗效等临床参数之间的关系,并进一步用Western Blot法分析了有髓外浸润和无髓外浸润AML患者HIF-1α蛋白表达水平。结果:HIF-1α在原代AML细胞中的表达量与年龄、性别、外周血白细胞计数、血红蛋白水平、血小板计数及外周血原始细胞比例等无关;不同FAB分型的AML患者的HIF-1α表达量无显著差异;标准1-2个疗程化疗后获得完全缓解(CR)的AML患者HIF-1α表达量低于2个疗程后未获得CR的AML患者,但无统计学差异;高表达HIF-1α的AML患者1年复发率要高于低表达者;伴有髓外浸润的AML患者的HIF-1α基因和HIF-1α蛋白表达量显著高于无髓外浸润的患者;HIF-1α表达水平与PI3K/AKT通路中重要的负调控分子PTEN水平呈明显负相关(r=-0.48,P=0.001)。结论:HIF-1α的表达与AML的髓外浸润和预后密切相关,有可能成为AML预后判断的一个参考指标。
Objective： This study was aimed to detect the expression of HIF-la in acute myeloid leukemia （AML） except acute promyelocyte leukemia （APL） and investigate the relationship of its expression levels with clinical parameters and prognosis. Methods： The primary AML cells were collected from peripheral blood of 53 newly diagnosed AML patients by using CD3 negative sorting. The expression of HIF-1α was measured by real-time fluorescent quantitative PCR （FQ-PCR） , and the relationship between expression level of HIF-1α and clinical parameters （age, sex, WBC count, clinical typing, prognosis） was analysed according to relative expression level. Furthermore, Western blot was used to detect the protein level of HIF-lct in AML patients with or without extramedullary infiltration. Results： The expression level of HIF-1α did not correlate with age, sex, WBC count, Hb level, Plt count and the percentage of blast. There was no significant difference of HIF-1α expression between different AML subtype based on FAB. The higher level of HIF-1α was found in AML patients who did not get complete remission after one or two courses of chemotherapy, however, the difference was not statistically significant. The relapse rate was higher in AML patients with the higher expression of HIF-1α. In addition, the higher level of HIF-1α mRNA and protein were found in bone marrow of AML patients with extramedullary infiltration （ P 〈 0.01 ）. The negative correlation between HIF-1α and PTEN was observed （ r=- 0.48, P = 0. 001 ）. Conclusions： Overexpression of HIF-1α are closely related with extramedullary infiltration and prognosis of acute myeloid leukemia, and may be used as an early indicator of extramedullary infiltration and prognosis.
出处
《中国实验血液学杂志》
CAS
CSCD
北大核心
2015年第1期19-23,共5页
Journal of Experimental Hematology
基金
国家自然科学基金(81100382)
国家和福建省临床重点专科建设项目资助
