The Effect Of Aminosteroid Derivatives On Human K562and MMG-olcell Lines Of Chronic Myelogenous Leukemia

Objects:To observe the effects of aminosteroid derivatives on human K562cell lines and MEG-01cell lines of chronic myelogenous leukemia on inhibition of proliferation and induction of differentiation, and further explore its mechanism.Methods:Liquid culture experiment and CCK-8kit was used to observe inhibition of cell proliferation ability of aminosteroid derivatives on K562and MEG-01cell lines; The CD71, CD41cell surface markers were detected by flow cytometry instrument to observe differentiation induction of aminosteroid derivatives on K562and MEG-01cell lines; Real-time RT-PCR experient was used to detect changes of mRNA expression of intracellular calcium ion channels TRPC1gene of aminosteroid derivatives on MEG-01cell lines and changes of mRNA expression of PTPN1gene on MEG-01cell lines.Results:According to the results of Cell counting and CCK-8kit, aminosteroid derivatives can effectively inhibit proliferation of K562and MEG-01cell lines, and it presented concentration dependence; According to the results of CCK-8kit, different aminosteroid derivatives all can effectively inhibit proliferation of MEG-01cell lines and their inhibition was not exactly the same. Wright staining after treatment for3d with aminosteroid derivatives at final concentration for10-6mol/L on K562and MEG-01cell lines found K562cells get wider, nuclear chromatin concentration coarsens, less nucleoplasm ratio, nuclear incisure, cytoplasm increased, light colored region appeared at nuclear cytoplasmic, which indicated that K562cells occured a certain degree of differentiation; MEG-01cells performed larger volume, nuclear chromatin concentration coarsens, less nucleoplasm ratio, irregular nucleus, cytoplasm increased, light colored region appeared at nuclear cytoplasmic, which indicated that MEG-01cells occured a certain degree of differentiation. Flow cytometry instrument analysis showed:after treatment for3d with aminosteroid derivatives at final concentration for10-6mol/L on K562and MEG-01cell lines, CD71-positive expression rate of K562and CD41-positive expression rate of MEG-01cells were increased. Results of real-time RT-PCR experient showed that the mRNA expression level of intracellular calcium ion channel TRPC1gene down-regulated and the mRNA expression level of PTPN1gene of MEG-01cell up-regulated after treatment for3d with aminosteroid derivatives at final concentration for10-6mol/L on MEG-01cell lines, of which RNA was extracted for reverse transcription and amplification.Conclusion:(1).Aminosteroid derivatives can effectively inhibit proliferation of K562and MEG-01cell lines, and it presented concentration dependence.(2).Aminosteroid derivatives can induce K562cell linestoword erythroid differentiation, inducing MEG-01toword megakaryocytic differentiation as well.(3).Aminosteroid derivatives can down-regulate mRNA expression level of intracellular calcium ion channel TRPC1genes of MEG-01cell lines.(4).Aminosteroid derivatives can up-regulate the mRNA expression level of PTPN1gene of MEG-01cells.