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Deficiency in myosin light-chain phosphorylation causes cytokinesis failure and multipolarity in cancer cells

Oncogene volume 29, pages 4183–4193 (2010)Cite this article

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Abstract

Cancer cells often have unstable genomes and increased centrosome and chromosome numbers, which are an important part of malignant transformation in the most recent model of tumorigenesis. However, very little is known about divisional failures in cancer cells that may lead to chromosomal and centrosomal amplifications. In this study, we show that cancer cells often failed at cytokinesis because of decreased phosphorylation of the myosin regulatory light chain (MLC), a key regulatory component of cortical contraction during division. Reduced MLC phosphorylation was associated with high expression of myosin phosphatase and/or reduced myosin light-chain kinase levels. Furthermore, expression of phosphomimetic MLC largely prevented cytokinesis failure in the tested cancer cells. When myosin light-chain phosphorylation was restored to normal levels by phosphatase knockdown, multinucleation and multipolar mitosis were markedly reduced, resulting in enhanced genome stabilization. Furthermore, both overexpression of myosin phosphatase or inhibition of the myosin light-chain kinase in nonmalignant cells could recapitulate some of the mitotic defects of cancer cells, including multinucleation and multipolar spindles, indicating that these changes are sufficient to reproduce the cytokinesis failures we see in cancer cells. These results for the first time define the molecular defects leading to divisional failure in cancer cells.

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Acknowledgements

We thank Dr David J Hartshorne (University of Arizona) for kindly providing the UPCI:SCC103 cell line and HEK293 with the MYPT1 expression cell line, respectively. We are also grateful to Dr Jeffrey D Hildebrand (University of Pittsburgh), Dr Anne R Bresnick (Albert Einstein College of Medicine), Dr Kathleen Kelly (National Cancer Institute), Dr David J Hartshorne (University of Arizona) and Dr James T Stull (University of Texas Southwestern Medical Center) for plasmids and antibodies. We also thank Dr Xiaojing Wang (University of Pittsburgh) for helpful discussions regarding statistical analysis and Dr Jian Huang (University of Texas Southwestern Medical Center) for helpful suggestions about urea glycerol electrophoresis. This study was supported in part by NIH grants R01DE016086 to WSS and SMG and R01DE14729 to SMG and WSS. Cytogenetic analyses were carried out in the UPCI Cytogenetics Facility, supported in part by NIH grant P30CA47904 to Ronald B Herberman/SMG.

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Authors and Affiliations

  1. Department of Biological Sciences, University of Pittsburgh, Pittsburgh, PA, USA

    Q Wu, R M Sahasrabudhe & W S Saunders

  2. Beckman Research Institute, City of Hope National Medical Center, Duarte, CA, USA

    L Z Luo

  3. Department of Human Genetics, University of Pittsburgh Graduate School of Public Health, Pittsburgh, PA, USA

    D W Lewis & S M Gollin

  4. The University of Pittsburgh Cancer Institute, Pittsburgh, PA, USA

    D W Lewis, S M Gollin & W S Saunders

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  1. Q Wu
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Correspondence to W S Saunders.

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Wu, Q., Sahasrabudhe, R., Luo, L. et al. Deficiency in myosin light-chain phosphorylation causes cytokinesis failure and multipolarity in cancer cells. Oncogene 29, 4183–4193 (2010). https://doi.org/10.1038/onc.2010.165

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