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Effects of Hydroxyl Morpholine Resistance on K562 Cells of Chronic Myelocytic Leukemia

??OBJECTIVE To investigate the inhibitory activity, induced differentiation-inducing activity and apoptosis-inducingactivity of hydroxyl morpholine(QDML-01) on chronic myelocytic leukemia cells line K562. METHODS The cell growth curve was drawn based on cell counting method. The IC₅₀ value of QDML-01 and positive control medicine to K562 cells were evaluated by methyl thiazolyl tetrazolium (MTT) assay method. Double soft agar assay method was carried out to study the ability of cell proliferation to determine efficacy of phamacognosy. The pathomorphism was analyed by the Wright-Giemsa staining method. The mechanism of cell apoptosis from morphology and gene level were investigated, by AO-EB double-staining method and DNA breakage test. The effect of QDML-01 on K562 cells from the protein level was determined by Western-blot. RESULTS The growth curves showed the K562 cells had strong cell vitality. They came into logarithmic phase on the third generation. The MTT assay RESULTS showed that the IC₅₀ values of QDML-01 and imatinib to K562 cells were 5.81 and 596.88 nmol??L^-1. Double soft agar colony formation test showed that clone formed at 21 d and the inhibitory rate of QDML-01 was 81.7%.It indicated that K562 cells were sensitive to QDML-01.Morphology test result showed that QDML-01 induced K562 cells to normal cells. The RESULTS of AO-EB double-staining method showed that QDML-01 induced the apoptosis of K562 cells. The study of DNA breakage test indicated that QDML-01 can induce the apoptosis of K562 cells to produce DNA banding with step-like. Western-blot analysis result suggested that QDML-01 can downregulated the expression of P210^bcr/abl protein. CONCLUSION QDML-01 has the inhibitory activity on chronic myelocytic leukemia cells line K562 by promoting the apoptosis of K562 cells and inducing differentiation to normal cells.