The Effects Of Aminosteroids On K562 And MEG-01 Cell Lines Of CML

Objective To explore the proliferation and differentiation-inducing effects and the potential mechanisms of aminosteroid on humam chronic myelogenous leukemia K562 and MEG-01 cell lines.Methods Proliferation was detected by MTT assay, differentiation was measured by morphology and flow cytometry.The changes of intracellular[Ca²⁺]were verified by fluorescence spectrophotometer.The changes of bcr/abl oncogene was testified by RT-PCR.Results 1 The effects of aminosteroid on proliferation and differentiation of K562 and MEG-01 cell lines.Proliferation:The growth of K562 and MEG-01 cell lines was inhibited after the treatment for 5d with 5 different aminosteroids at the concentration of 10^-5mol/L.The inhibition rates for K562 cells in MTT assay were respectively as follows:Z-30(62.18±4.72) %,Z-31(35.25±2.16)%,Z-32(42.86±7.32)%,Z-33(64.71±1.24) %,Z-34(73.95±8.82)%;and the inhibition rates for MEG-01 cells in MTT assay were respectively as follows:Z-30(43.24±5.61) %,Z-31(45.65±3.24)%,Z-32(60.81±2.57)%,Z-33(66.22±6.26) %,Z-34(72.97±7.28).Differentiation:The morphlogy showed the differentiation tendency after the treatment of K562 and MEG-01 cells.Data from flow cytometry showed that 5 kind of aminosteroids(Z-30,31,32,33,34) induced K562 cells toward erythroid differentiation at 10^-6mol/L after cultured for 5d,in which the antigen CD71 expression rates were as follows:91.63%,91.07%,85.16%,80.85%,75.51%;Data from flow cytometry showed that 5 kind of aminosteroids(Z-30,31,32,33,34) induced MEG-01 cells toward megakaryocytic differentiation at 10^-6mol/L after cultured for 5d,in which the antigen CD41 expression rates were as follows:74.7%,64.5%,64.58%,54.96%,59.65%.2 The effects of aminosteroids on intracellular[Ca²⁺]in K562 and MEG-01 cells:Data from fluorospectrophotometer showed that,5 kind of aminosteroids(Z-30,31,32,33,34) could all lowered the total intracellular[Ca²⁺]in K562 cells at 10^-6mol/Lafter cultured for 30 min;and the same effect was observed on MEG-01 cells after cultured for 1h.3 The effects of aminosteroid on bcr/abl oncogene of K562 and MEG-01 cell lines:After treated for 5d,it was found that all 5 kind of aminosteroids(Z-30,31,32,33,34)down-regulated the bcr/abl oncogene expression at 10^-6mol/L.Conclution1 All of the 5 aminosteroids inhibited the proliferation of K562 and MEG-01 cells,but the sensitivities were different for different aminosteroids. 2 All aminosteroids induced K562 cells toward erythroid differentiation,and induced MEG-01 cells toward megakaryocytic differentiation,but the differentiation degree varies depending on different aminosteroids.3 All aminosteroids decreased intracellular[Ca²⁺]in K562 and MEG-01 cells,and this may be related to their blocking effects on calcium channels.And what's more,the reduction of intracellular[Ca²⁺] may connect to the mechanism of the aminosteroids's effect of inhibiting proliferation and inducing differentiation.4 All aminosteroids down-regulated the bcr/abl oncogene both in K562 and MEG-01 cell lines,but the degrees of the down-regulation were different depending on different aminosteroids.