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《Journal of Beijing Medical University》 2000-01
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Detection of DNA-methyltransferase activity of leukemia cells with radiology microassay

BAI Zhi Yong 1, XU Guo Bin 2, WU Shu Lan 1 (1. Department of Hematology, 2. Department of Laboratory, the First Hospital, Beijing Medical University, Beijing 100034, China)  
Objective:To establish a sensitive radiology microassay technique for studying activity of DNA methyltransferase in leukemia cells, and evaluate its feasibility. Methods: 1.1×10 8 Bq 3H SAM used as a methyl donor and 0.5 μg Poly[d(I C)d(I C)] as a receptor, MTase activity in mononucleated cells from bone marrow of patients with acute leukemia and of control subjects with no hematological malignancies were detected. cpm were counted with 5 μg cellular protein incubated for 2 hours. All detection was performed in duplicates, each experiment had positive control with HL 60 cells and negative control without Poly [d(I C)d(I C)]. Results: The method was linear up form 700 to 20 000 cpm when the quality of cellular proteins was between 0.25 μg and 7.5 μg, Y cpm = 2 864 X + 142, r =0.998.Within group and day to day variations were 5.9% and 11.4% respectively. The sensitivity of the method was 2.4×10 4 Bq. Mean MTase activity in 15 cases with acute leukemia was (9.1±4.6)%,4 fold higher than that in 11 cases with no hematological malignancies which was ( 2.1± 1.6)% ( P 0.05); and the value from 11 acute leukemia patients in complete remission was ( 4.1± 2.6)%, which was in the same range as the control group. Conclusion:Radiology microassay had high sensitivity and good replication for detecting MTase activity of leukemia cells. MTase activity of patients with acute leukemia increased, and it decreased in thesame range as control group when thecomplete remission was achieved.
【Fund】: 卫生部科学研究基金! ( 94 1 2 5 0 )
【CateGory Index】: Q555
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