人端粒重复序列结合因子(hTRF1)蛋白质在急性白血病中的表达水平与端粒酶活性关系的研究

Expression Level of TRF1 Protein in Human Acute Leukemia and Its Relationship with Activity of Telomerase

为了研究人端粒重复序列结合因子(TRF1)蛋白质在急性白血病(AL)及正常骨髓组织中的表达水平,AL和正常骨髓组织中端粒酶活性及TRF1蛋白质表达水平与端粒酶活性的关系,定量分析了AL患者及正常人骨髓组织中TRF1蛋白质的表达水平,并应用经倍比稀释的TRF133-277纯化蛋白质作为定量标准,建立以抗TRF133-277单克隆抗体的定量Westernblot的方法,而且以该方法检测TRF1蛋白质在组织中的表达水平;另外,应用TRAP-PCR-ELISA法检测AL患者及正常人骨髓组织端粒酶活性,以研究TRF1蛋白质表达水平与端粒酶活性的关系。结果表明:20例AL患者骨髓组织中TRF1表达水平较正常人骨髓组织显著降低,差异具有显著性(P<0.01);急性淋巴细胞性白血病患者的TRF1表达水平较急性非淋巴细胞性白血病患者略减低,但差异无统计学意义(P>0.05);化疗缓解的患者TRF1表达水平较前增高,但仍低于正常(P<0.01);化疗后完全缓解患者的TRF1表达水平较未缓解的患者显著增高,差异具有显著性(P<0.01);AL患者初发未治时骨髓细胞端粒酶的活性明显高于正常人(P<0.05)和首次缓解者(P<0.01);初发未治时,ALL患者骨髓细胞端粒酶活性略高于ANLL患者,但差异无统计学意义(P>0.05)。TRF1蛋白质表达水平与端粒酶活性呈明显负相关(P<0.001)。结论:TRF1蛋白质在AL患者骨髓细胞中的表达明显减低,且与疗效及端粒酶活性相关。

The study was aimed to investigate the expression level of TRF1 protein in human acute leukemia and relationship between expression level of TRF1 protein and activity of telomerase. A quantitative Western blot technique was developed using anti-TRF1^33-277 monoclonal antibody and GST-TRF1 fusion protein as a standard to further determine the expression level of TRF1 protein in total proteins extracted from clinical specimens. 20 cases of acute leukemias were studied when 11 normal volunteer＇s bone marrow was used as control. The results showed that the expression level of TRF1 protein in normal bone marrow （2.217 ±0.461 μg/μl） was significantly higher than that in bone marrow of acute leukemia patients （0. 754 ± 0.343 μg/μl） （ P 〈 0.01 ）. There was no remarkable difference of expression level of TRF1 protein between ALL and ANLL （0.628 ±0. 281 μg/μl vs 0. 844 ±0. 360 μg/μl, P 〉0.05）. After chemotherapy, TRF1 expression level of patients with complete remeision raised （0.772 ±0. 307 μg/μl vs 1. 683 ±0.344 μg/μl, P 〈 0.01 ）, but lower than that of normal （ 2. 217 ±0.461 μg/μl , P 〈 0.01 ）. TRF1 expression level of patients without complete remission was not remarkable different after chemotherapy （0. 726 ±0.443 μg/μl vs 0. 894 ±0.338 μg/μl, P 〉 0.05）. TRF1 expression level of patients with complete remession was higher than that in patients without complete remession （1.683 ±0.344 μg/μl vs 0.894 ±0.338 μg/μl, P〈0. 01）. For all sample the telomerase activity was determined. It was confmned that the activity of telomerase in normal bone marrow was lower than that in bone marrow of acute leukemia patients （0. 125 ±0.078 μg/μl vs 0.765 ±0.284 μg/μl, P〈0.01）. There was no significatly difference of expression level of TRF 1 protein between ALL and ANLL （ 0.897 ± 0.290 μg / μl vs 0.677 ±0.268 μg / μl, P 〉 0.05）. After chemotherapy, telomerase activity of patients with complete remeision reduced （0.393 ±0. 125 μg/μl）, but higher than that of normal （0. 125 ±0. 078 μg/μl, P 〈0. 01 ）. It is concluded that expression level of TRF1 protein in AL patients is significantly decrese and associated with therapeutic efficaciousness and the activity of telomerase （P 〈0. 001 ）.