Silencing of RB1 but not of RB2/P130 induces cellular senescence and impairs the differentiation potential of human mese
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Cellular and Molecular Life Sciences
RESEARCH ARTICLE
Silencing of RB1 but not of RB2/P130 induces cellular senescence and impairs the differentiation potential of human mesenchymal stem cells Nicola Alessio · Wolfgang Bohn · Verena Rauchberger · Flavio Rizzolio · Marilena Cipollaro · Michael Rosemann · Martin Irmler · Johannes Beckers · Antonio Giordano · Umberto Galderisi
Received: 27 June 2012 / Revised: 24 November 2012 / Accepted: 26 November 2012 / Published online: 31 January 2013 © Springer Basel 2013
Abstract Stem cell senescence is considered deleterious because it may impair tissue renewal and function. On the other hand, senescence may arrest the uncontrolled growth of transformed stem cells and protect organisms from cancer. This double function of senescence is strictly linked to the activity of genes that the control cell cycle such as the retinoblastoma proteins RB1, RB2/P130, and P107. We took advantage of the RNA interference technique to analyze the role of these proteins in the biology of mesenchymal stem cells (MSC). Cells lacking RB1 were prone to DNA damage. They showed elevated levels of p53 and p21cip1 and increased regulation of RB2/P130 and P107 expression. These cells gradually adopted a senescent phenotype with impairment Electronic supplementary material The online version of this article (doi:10.1007/s00018-012-1224-x) contains supplementary material, which is available to authorized users. N. Alessio · M. Cipollaro · U. Galderisi Department of Experimental Medicine, Biotechnology and Molecular Biology Section, Second University of Naples, Naples, Italy W. Bohn · V. Rauchberger Department of Tumorvirology, Heinrich-Pette-Institute, Leibniz-Institute for Experimental Virology, Hamburg, Germany F. Rizzolio · A. Giordano · U. Galderisi (*) Sbarro Institute for Cancer Research and Molecular Medicine, Center for Biotechnology, Temple University, 1900 North 12th Street, Philadelphia, PA 19107-6799, USA e-mail: [email protected]; [email protected] M. Rosemann Helmholtz Zentrum, National Research Center for Environment and Health, GmbH, Institute of Radiation Biology, Munich, Germany
of self-renewal properties. No significant modification of cell growth was observed as it occurs in other cell types or systems. In cells with silenced RB2/P130, we detected a reduction of DNA damage along with a higher proliferation rate, an increase in clonogenic ability, and the diminution of apoptosis and senescence. Cells with silenced RB2/P130 were cultivated for extended periods of time without adopting a transformed phenotype. Of note, acute lowering of P107 did not induce relevant changes in the in vitro behavior of MSC. We also analyzed cell commitment and the osteo-chondroadipogenic differentiation process of clones derived by MSC cultures. In all clones obtained from cells with silenced retinoblastoma genes, we observed a reduction in the ability to differentiate compared with the control clones. In summary, our data show evidence that the silencing of the expression of RB1 or RB
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