原创力文档role of cyclin b1cdc2 up-regulation in the development of mitotic prometaphase arrest in human breast cancer cells treated with nocodazole细胞周期蛋白的作用b1cdc2老年病的发展在人类乳腺癌细胞有丝分裂前中期逮捕与诺考达唑治疗.pdfVIP
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role of cyclin b1cdc2 up-regulation in the development of mitotic prometaphase arrest in human breast cancer cells treated with nocodazole细胞周期蛋白的作用b1cdc2老年病的发展在人类乳腺癌细胞有丝分裂前中期逮捕与诺考达唑治疗
Role of Cyclin B1/Cdc2 Up-Regulation in the
Development of Mitotic Prometaphase Arrest in Human
Breast Cancer Cells Treated with Nocodazole
Hye Joung Choi, Masayuki Fukui, Bao Ting Zhu*
Department of Pharmacology, Toxicology and Therapeutics, School of Medicine, University of Kansas Medical Center, Kansas City, Kansas, United States of America
Abstract
Background: During a normal cell cycle, the transition from G2 phase to mitotic phase is triggered by the activation of the
cyclin B1-dependent Cdc2 kinase. Here we report our finding that treatment of MCF-7 human breast cancer cells with
nocodazole, a prototypic microtubule inhibitor, results in strong up-regulation of cyclin B1 and Cdc2 levels, and their
increases are required for the development of mitotic prometaphase arrest and characteristic phenotypes.
Methodology/Principal Findings: It was observed that there was a time-dependent early increase in cyclin B1 and Cdc2
protein levels (peaking between 12 and 24 h post treatment), and their levels started to decline after the initial increase. This
early up-regulation of cyclin B1 and Cdc2 closely matched in timing the nocodazole-induced mitotic prometaphase arrest.
Selective knockdown of cyclin B1or Cdc2 each abrogated nocodazole-induced accumulation of prometaphase cells. The
nocodazole-induced prometaphase arrest was also abrogated by pre-treatment of cells with roscovitine, an inhibitor of
cyclin-dependent kinases, or with cycloheximide, a protein synthesis inhibitor that was found to suppress cyclin B1 and
Cdc2 up-regulation. In addition, we found that MAD2 knockdown abrogated nocodazole-induced accumulation of cyclin B1
and Cdc2 proteins, which was accompanied by an attenuation of nocodazole-induced prometaphase arrest.
Conclusions/Significance: These observations demonstrate that the strong
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