A Study On Apoptosis Procedure And Molecular Mechanism Of Leukemia Cells Induced By TGP

Purpose:For Experimental exploring the inhibition effects on K562?HL60 and fresh bone marrow leukemia cells,and observing apoptosis procedure of K562?HL60?and fresh bone marrow leukemia cells cultured with TPG.And to study the changes of HL60 cultured with TGP on morphology,gene and protein expression,verify the apoptosis process,and make a preliminary inference of the way of cell apoptosis.Methods: Build cells models of K562 and HL60 which was in logarithmic period cultured with TGP.The leukemia cells was extracted with lymphocyte wseparation medium from fresh of in the bone marrow leukemia patients,observe the cell proliferation with CCK8 method?Study on the effects of inhibition rate of leukemia cells with these factor,such as concentration of TGP?culturing time and leukemia cell type.Study on HL60 cultured with 200 ug/ml TGP and observe the morphological changes by microscopy.Study changes of endogenous apoptosis factor,such as the Bcl-2,Bax and Caspase3 and Caspase9.Results:(1)Accompanied with the increasing of culturing time of K562 and HL60 and human fresh bone marrow leukemia cells with TGP,results showed inhibition rate constantly rise;accompanied with the rising of the concentrations of TGP,inhibition rate was also rising;Under the condition of 72 hours of training the IC50 value concentration respectively was 25.638±1.409?19.04±1.28 and16.317±1.213.Through three factor analysis of variance,the result showed that the concentration of TGP? types of leukemia stem cells?culturing time on inhibition rate all had significant effects(P < 0.05),the factor of TGP concentration had the largest F value performance,it reached 1629.132.this indicated that its effect was more intense.On the effect of two factorinteractions,TGP × leukemia stem cell type,concentration of TGP×incubation time for leukemia stem cells all had significant inhibition effect(P< 0.01).And on the three factors interaction effect,F value was 0.472 and had not significant.(2)Accompanied with the extension of culturing time,the cell broken more serious morphologically;Observed the cells witch was cultured with 12 h,HL60 cells had obvious morphological characteristics changes such as edge set,cataclastic typical apoptotic nuclei pyknosis,the nucleus shape on the irregular degree has been increased.Using flow cytometry instrument to observe the apoptosis rate,the results showed that the K562 apoptosis rate rose from 3.0% to 42.49%;The HL60 rose from 3.87% to 46.73%;fresh bone marrow leukemia stem cells rose from 4.15% to 52.46%,the consequence indicated that it had significantly increasing(P < 0.05).(3)on mRNA expression level,with the increase of incubation time,the expression of Bcl-2levels falling,Bax,Caspase3 and Caspase9 rising;On the Bcl-2 / Bax ratio,12 h,starting from the blank control group to develop its value,the phenomenon showed a declining trend.On the protein expression level,it also showed the Bcl-2 expression level falling,Bax,Caspase3 and Caspase9 rising;Over the Bcl-2 / Bax ratio,and it also showed the same trend as the former.Conclusion:K562 and HL60 and fresh bone marrow leukemia cells cultured with TGP had outstanding inhibiting effects,and always accompanied with outstanding time dependent and dose dependent.Therefore,combined with the conclusions above,the study believe that TGP has a great advantage compared with the traditional chemotherapy drugs,regulation of setting the appropriate concentration of TGP,at the same time considering that accompanied with other drugs should have a synergistic effect,increasing the therapeutic effect,at the same time also can eliminate the related side effects,make the TGP becoming an excellent clinical leukemia treatment.