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目的:建立一种实时荧光RT-PCR方法,定量检测急性髓细胞性白血病(AML)患者外周血白细胞细胞TRF1、TRF2和hRAP1mRNA的表达水平及相互关系。方法:选取48例初治AML患者和21例健康志愿者,采用实时荧光RT-PCR与Lightcyeler荧光PCR仪定量检测TRF1、TRF2和hRAP1mRNA的表达水平;采用2-△△Ct法处理实时荧光定量RT-PCR数据。结果:AML首治患者TRF1、TRF2和hRAP1mRNA表达水平较对照组健康志愿者分别下降了2.9倍,2.2倍和16.6倍,且差异有统计学意义;AML患者组TRF1和TRF2mR-NA的表达改变具有一定的相关性(r=0.57,P<0.01)。结论:成功利用患者外周血检测出端粒结合蛋白TRF1,TRF2在白血病细胞中mRNA的表达水平下降,并首次检测发现hRAP1在AML患者细胞中存在表达下降,间接说明了hRAP1的表达下降也是白血病的发病原因之一。
OBJECTIVE: To establish a real-time fluorescence RT-PCR method for the quantitative detection of the expression of TRF1, TRF2 and hRAP1 mRNA in peripheral blood leukocytes of patients with acute myeloid leukemia (AML) and their correlation. Methods: Forty-eight newly diagnosed AML patients and 21 healthy volunteers were enrolled in this study. The expression of TRF1, TRF2 and hRAP1 mRNA were detected by real-time fluorescent RT-PCR and Lightcyeler fluorescence PCR. The real-time fluorescent quantitative RT -PCR data. Results: The expression of TRF1, TRF2 and hRAP1 mRNA in AML patients decreased by 2.9-fold, 2.2-fold and 16.6-fold respectively compared with healthy volunteers in the control group, and the difference was statistically significant. The changes of TRF1 and TRF2mR- A certain correlation (r = 0.57, P <0.01). CONCLUSIONS: The expression of telomere binding protein TRF1 and TRF2 in leukemic cells has been successfully detected using peripheral blood of patients. The expression of hRAP1 in the cells of AML patients decreased after the first detection, which indirectly indicates that the decrease of hRAP1 expression is also leukemia One of the causes.