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MJ‐29 interacts with microtubule polymerization and induces mitotic arrest following apoptosis in human leukemia U937 cell line
Author(s) -
Yang JaiSing,
Chung JingGung,
Hour ManJan,
Wang ShuFang,
Lin KueiLi
Publication year - 2007
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.21.6.a794
Subject(s) - apoptosis , mitosis , u937 cell , dapi , microbiology and biotechnology , annexin , cell cycle , chemistry , xiap , cell growth , cell culture , cyclin dependent kinase 1 , caspase , biology , programmed cell death , biochemistry , genetics
In this study, MJ‐29(6‐pyrrolidinyl‐2‐(2‐substituted phenyl)‐4‐quinazolinones derivatives) was examined for its growth inhibition effect on U937 human leukemia cell line. After 24 hours treatment of MJ‐29,PI incorporation was applied to determine the proliferation rate. A dose and time‐dependent decrease in cell proliferation was observed. The cytotoxic effect of MJ‐29 on normal cells (HUVEC and peripheral mononuclear cells) was lower than that on U937 cells of treatment. Cell cycle analysis of U937 cells showed that MJ‐29 induced significant G2/M arrest and apoptosis. Immuno‐fluorescence microscopy and in vitro tubulin assembly assays indicated that MJ‐29 binding to tubulin and disrupts microtubule organization. We found that MJ‐29 up‐regulated the protein levels of p161‐CDK1,Chk1,Chk2,p21 and increased cyclin B and CDK1 kinase activity. MJ‐29‐treated U937 cells presented typical apoptosis by DAPI staining, increase of Annexin V binding, internucleosomal DNA cleavage. In addition, release of cytosolic cytochrome c¡BPro‐caspase‐9¡BApaf‐1 and AIF, down‐regulation of Bcl‐2, Bcl‐xL¡B xIAP¡BcIAP‐1¡BcIAP‐2 and up‐regulation of phospho‐Bcl‐2¡BBax¡BBad¡BBak were detected by western blotting. Caspases activity assay and Sub‐G1 nuclei analysis of MJ‐29‐treated U937 indicated that the MJ‐29 induced apoptosis was mainly mediated by activation of caspase 9 and 3. Our results indicated that the ability of MJ‐29 to inhibit cell proliferation may be mediated by both the induction of mitotic arrest and apoptosis. μ