Silencing of RB1 but not of RB2/P130 induces cellular senescence and impairs the differentiation potential of human mesenchymal stem cells

被引:55
作者
Alessio, Nicola [1 ]
Bohn, Wolfgang [2 ]
Rauchberger, Verena [2 ]
Rizzolio, Flavio [3 ]
Cipollaro, Marilena [1 ]
Rosemann, Michael
Irmler, Martin [4 ]
Beckers, Johannes [4 ,5 ]
Giordano, Antonio [3 ,6 ,7 ]
Galderisi, Umberto [1 ,3 ,6 ]
机构
[1] Univ Naples 2, Dept Expt Med, Biotechnol & Mol Biol Sect, Naples, Italy
[2] Leibniz Inst Expt Virol, Heinrich Pette Inst, Dept Tumorvirol, Hamburg, Germany
[3] Temple Univ, Sbarro Inst Canc Res & Mol Med, Ctr Biotechnol, Philadelphia, PA 19107 USA
[4] Natl Res Ctr Environm & Hlth GmbH, Helmholtz Zentrum, Inst Expt Genet, Munich, Germany
[5] Tech Univ Munich, Ctr Life & Food Sci Weihenstephan, Chair Expt Genet, WZW, Freising Weihenstephan, Germany
[6] Human Hlth Fdn, Spoleto, Italy
[7] Univ Siena, Dept Med Sci Surg & Neurosci, I-53100 Siena, Italy
关键词
Retinoblastoma gene family; Marrow stromal stem cells; Differentiation; Apoptosis; Senescence; Cell cycle; DNA damage; BONE-MARROW; RETINOBLASTOMA PROTEIN; ADIPOCYTE DIFFERENTIATION; REDOX REGULATION; FAMILY-MEMBERS; STROMAL CELLS; DNA-REPAIR; IN-VITRO; EXPRESSION; DAMAGE;
D O I
10.1007/s00018-012-1224-x
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
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 p21(cip1) and increased regulation of RB2/P130 and P107 expression. These cells gradually adopted a senescent phenotype with impairment 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-chondro-adipogenic 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 RB2/P130 is not compensated by other gene family members, and this profoundly affects MSC functions.
引用
收藏
页码:1637 / 1651
页数:15
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