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Anoikis triggers Mdm2-dependent p53 degradation

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Abstract

The extracellular matrix (ECM) plays a key role in cell–cell communication and signaling, and the signals it propagates are important for tissue remodeling and survival. However, signals from disease-altered ECM may lead to anoikis—apoptotic cell death triggered by loss of ECM contacts. Previously, we found that an altered fibronectin matrix triggers anoikis in human primary ligament cells via a pathway that requires p53 transcriptional downregulation. Here we show that this p53 reduction is suppressed by transfecting cells with Mdm2 antisense oligonucleotides or small interfering RNA. Similar results were found in cells treated to prevent p53 and Mdm2 interactions. When p53 was overexpressed in cells lacking Mdm2 and p53, p53 levels were unaffected by anoikis conditions. However, cells cotransfected with p53 and wild type Mdm2, but not a mutant Mdm2, exhibited decreased p53 levels in response to anoikis conditions. Thus, cells under anoikis conditions undergo p53 degradation that is mediated by Mdm2.

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Notes

  1. The abbreviations used are: ECM, extracellular matrix; cAFn and AFn, alternatively spliced V region (V+) and containing intact (H+) or a mutated, nonfunctional (H−) high-affinity heparin binding domain; TBST, 25 mM Tris, 150 mM NaCl, 0.05% Tween-20.

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Acknowledgments

We thank Dr. Gerard P. Zambetti (St. Jude Children’s Research Hospital, Memphis, TN) for p53-null fibroblasts, Dr. Gigi Lozano (University of Texas M.D. Anderson Cancer Center, Houston, TX) for p53/Mdm2-null fibroblasts, Dr. Bert Vogelstein (Johns Hopkins University School of Medicine, Baltimore, MD) for human wildtype p53 and human Mdm2 pCMV expression plasmids, Dr. Arnold J. Levine (Princeton University, Princeton, NJ) for human Mdm2 pCHDMΔ222-437 expression plasmid, Dr. Paul W. Johnson for mentoring and the fibronectin fragments and Stephen Ordway for editorial assistance. This study was support by an NIH grant RO1 DE013725 (to Y. L. K.).

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  1. Department of Periodontics and Oral Medicine, University of Michigan, School of Dentistry, 1011 N. University Ave., Ann Arbor, MI, 48109-1078, USA

    Abhijit Ghosh, Tina Chunyuan Chen & Yvonne L. Kapila

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  1. Abhijit Ghosh
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  2. Tina Chunyuan Chen
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Correspondence to Yvonne L. Kapila.

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Ghosh, A., Chen, T.C. & Kapila, Y.L. Anoikis triggers Mdm2-dependent p53 degradation. Mol Cell Biochem 343, 201–209 (2010). https://doi.org/10.1007/s11010-010-0514-6

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