通过实时逆转录聚合酶链反应检测急性髓性白血病中hTERT信使核糖核酸水平。

hTERT mRNA levels by real-time RT-PCR in acute myelogenous leukemia.

作者信息

Huh Hee Jin, Huh Jung Won, Yoo Eun Sun, Seong Chu Myong, Lee Miae, Hong Ki Sook, Chung Wha Soon

机构信息

Department of Laboratory Medicine, Ewha Womans University, College of Medicine, Seoul, South Korea.

出版信息

Am J Hematol. 2005 Aug;79(4):267-73. doi: 10.1002/ajh.20394.

DOI:10.1002/ajh.20394

PMID:16044449

Abstract

The purpose of this study was to investigate whether levels of hTERT mRNA, as determined by real-time RT-PCR, are associated with prognosis and clinical course in AML patients. Fifty-four bone marrow specimens from 21 patients diagnosed with de-novo AML were included. The level of hTERT mRNA was measured with the Telo TAGGG hTERT Quantification Kit (Roche Diagnostics, Mannheim, Germany), using a LightCycler Instrument (Roche Diagnostics). The level of hTERT mRNA was determined as the relative ratio (RR), which was calculated by dividing the level of hTERT mRNA by the level of the porphobilinogen deaminase (PBGD) housekeeping gene in the same samples [1,000x(hTERT/PBGD)]. The expression rates of hTERT mRNA were significantly higher at diagnosis (73%) and during relapse (80%) than during remission (27%) (P<0.05). The median RR for diagnosis or relapse was significantly higher than that for patients in remission (P<0.05). hTERT mRNA expression was not correlated with CD34 expression, blast counts, white blood cell counts, or chromosomal abnormality (P>0.05). Two patients who showed hTERT mRNA expression during remission (RR 3.14 and 7.15, respectively) relapsed after 1 month. Among seven patients with high hTERT mRNA levels (RR>9.51), 4 failed to achieve complete remission (CR), whereas 4 of 5 patients without hTERT mRNA expression at diagnosis or during relapse achieved CR (P>0.05). Patients showing a trend of increasing hTERT mRNA levels failed to reach a second CR after relapse, while those with a trend toward decreasing hTERT mRNA did achieve CR. Among eight samples showing hTERT mRNA expression in remission (RR>0), 5 were obtained from patients who had received GCSF within 14 days. The expression rate and level of hTERT mRNA during remission were significantly higher in patients who had previously received GSCF (56%, RR=0.15) than in other patients (15%, RR=0) (P<0.05). Serial and quantitative analysis of hTERT mRNA may be a useful marker for prediction of prognosis and monitoring in AML patients.

摘要

本研究的目的是调查通过实时逆转录聚合酶链反应（RT-PCR）测定的人端粒酶逆转录酶（hTERT）mRNA水平是否与急性髓系白血病（AML）患者的预后及临床病程相关。纳入了21例诊断为初发性AML患者的54份骨髓标本。使用罗氏诊断公司（德国曼海姆）的Telo TAGGG hTERT定量试剂盒，通过LightCycler仪器（罗氏诊断公司）测量hTERT mRNA水平。hTERT mRNA水平以相对比率（RR）确定，其计算方法是将同一样本中hTERT mRNA水平除以胆色素原脱氨酶（PBGD）管家基因的水平[1,000×（hTERT/PBGD）]。hTERT mRNA的表达率在诊断时（73%）和复发时（80%）显著高于缓解期（27%）（P<0.05）。诊断或复发时的RR中位数显著高于缓解期患者（P<0.05）。hTERT mRNA表达与CD34表达、原始细胞计数、白细胞计数或染色体异常均无相关性（P>0.05）。两名在缓解期显示hTERT mRNA表达的患者（RR分别为3.14和7.15）在1个月后复发。在7例hTERT mRNA水平高（RR>9.51）的患者中，4例未达到完全缓解（CR），而在诊断或复发时无hTERT mRNA表达的5例患者中有4例达到CR（P>0.05）。hTERT mRNA水平呈上升趋势的患者复发后未达到第二次CR，而hTERT mRNA呈下降趋势的患者则达到了CR。在8份缓解期显示hTERT mRNA表达（RR>0）的样本中，5份来自在14天内接受过粒细胞集落刺激因子（GCSF）的患者。既往接受过GCSF的患者缓解期hTERT mRNA的表达率和水平显著高于其他患者（56%，RR=0.15）（15%，RR=0）（P<0.05）。hTERT mRNA的系列定量分析可能是预测AML患者预后和进行监测的有用标志物。