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目的利用实时荧光定量RT-PCR方法,检测白血病细胞人类端粒酶反转录酶(hTERT)的表达水平并探讨其临床意义。方法采用基于TaqMan荧光技术的实时定量RT-PCR方法检测白血病患者骨髓单个核细胞及10种白血病细胞系hTERT mRNA拷贝数,来自同种基因移植供者的正常人骨髓细胞作为对照。结果与28例正常人组相比,33例AML1/ETO融合基因阳性的急性髓性白血病(AML)患者骨髓细胞hTERT mRNA水平较高,中位数分别为8.96%(0.23%～76.73%)和4.20%(0.21%～19.90%)(P<0.01)。47例B-ALL患者骨髓细胞hTERT mRNA水平中位数为20.80%(0.25%～1566.34%),高于正常人组(P<0.001)及156例AML组(中位数为2.06%,范围为0～2133.41%)。70例慢性髓性白血病(CML)、36例M3型白血病患者hTERT mRNA水平中位数分别为0.57%(0～14.77%)、0.53%(0～11.10%),均低于正常人组(P均<0.001)。15例治疗后达到完全缓解的急性白血病患者与初治时相比,hTERT mRNA水平明显下降,中位数分别为46.53%(8.96%～218.10%)和2.25%(0.18%～45.85%)(P<0.001)。10种白血病细胞系的hTERT mRNA水平差异甚大。结论 hTERT mRNA在人白血病细胞中的表达水平存在异质性。高表达hTERT mRNA可能是B-ALL及AML1/ETO阳性的AML发病机制中的因素之一。
Objective To detect the expression level of human telomerase reverse transcriptase (hTERT) in leukemia cells by real-time fluorescent quantitative RT-PCR and to explore its clinical significance. Methods Real-time quantitative RT-PCR based on TaqMan fluorescence was used to detect hTERT mRNA copy number in bone marrow mononuclear cells and leukemia cell lines from leukemia patients. Normal human bone marrow cells from allogeneic donor were used as control. Results Compared with 28 normal controls, 33 patients with acute myeloid leukemia (AML) who had AML1 / ETO fusion gene had higher levels of hTERT mRNA in bone marrow cells with a median of 8.96% (0.23% -76.73%) and 4.20% (0.21% ~ 19.90%) (P <0.01). The median hTERT mRNA level in bone marrow cells of 47 patients with B-ALL was 20.80% (0.25% -1566.34%), higher than that in normal people (P <0.001) and 156 patients with AML (median 2.06% 0 to 2133.41%). The median hTERT mRNA levels in 70 patients with chronic myeloid leukemia (CML) and 36 patients with M3 leukemia were 0.57% (0-14.77%) and 0.53% (0-11.10%), respectively, which were lower than those in normal subjects (P All <0.001). HTERT mRNA levels were significantly decreased in 15 patients with acute leukemia who achieved complete remission after treatment (median, 46.53%, 8.96% ~ 218.10%, 2.25%, 0.18% ~ 45.85%, P <0.001). The hTERT mRNA levels of 10 leukemia cell lines vary widely. Conclusion The expression level of hTERT mRNA in human leukemia cells is heterogeneous. High expression of hTERT mRNA may be one of the factors in the pathogenesis of B-ALL and AML1 / ETO-positive AML.