Effects of inducing apoptosis on HL-60 cells with Mitoxantrone / 中国免疫学杂志

ABSTRACT

Objective:To investigate the mechanisms of Mitoxantrone(MTN) induced apoptosis in human acute myloid leukemia HL 60 cells.Methods:Exposured log phase growth HL 60 cells to different Mitoxantrone concentrations for different time respectively and then test the inhibitive effect by modified tertrozalium salt(MTT) assay.Morphologic evidence for apoptosis of MTN induced on HL 60 cells was determined by transmission electron microscope.The detection of MTN induced internucleosamal DNA fragmentation by agurose gele lectrophoresis.The effects on HL 60 cells proteins(Bcl 2,Bax) implicated in apoptosis were determined by Flow cytoetric.Results:(1)The antiproliferative effects were observed following exposure micromilligram level of MTN.It shows a very substantial differences in the dose and duration dependency of the observed antiproliferative and cytotoxic effects.(2)The HL 60 cells treated by MTN are observed apoptosis characteristic morphological changes such as complete cell membrane,nuclear fragmention apoptosic bodies and found about 200 bp trapezoidal belt with DNA electrophoresis.(3)The protein test results of Bcl 2 and Bax indicate that when the time of MTN treating HL 60 cells is the same,Bcl 2 protein and the ratio of Bcl 2 protein and Bax protein(Bcl 2/Bax) descend along with the ascending concentration of MTN.Bax protein has no connection with the concentration of MTN.When the concentration of MTN treating HL 60 cells are the same.The ratio of Bcl 2/Bax descends as the time of MTN treating HL 60 cells is prolonged.Conclusion:MTN has the function of inducing apoptosis on HL 60 cells with descent of Bcl 2 and Bcl 2/Bax.The regulation pathway of Bcl 2 may be a mechanism of MTN induced apoptosis on HL 60 cells.