Vol.:(0123456789) 1 3 Molecular Genetics and Genomics https://doi.org/10.1007/s00438-018-1495-5 ORIGINAL ARTICLE Proliferation of aneuploid cells induced by CENP-E depletion is counteracted by the p14 ARF tumor suppressor Lorena Veneziano 1  · Viviana Barra 1,2  · Danilo Cillufo 1  · Aldo Di Leonardo 1,3 Received: 23 January 2018 / Accepted: 24 September 2018 © Springer-Verlag GmbH Germany, part of Springer Nature 2018 Abstract The spindle assembly checkpoint (SAC) is a cellular surveillance mechanism that ensures the fdelity of chromosomes segregation. Reduced expression of some of its components weakens the SAC and induces chromosome instability and ane- uploidy, which are both well-known hallmarks of cancer cells. Centromere protein-E (CENP-E) is a crucial component of the SAC and its function is to facilitate kinetochore microtubule attachment required to achieve and maintain chromosome alignment. The present study investigates the possible role of p14 ARF as a controller of aneuploid cells proliferation. We used RNA interference to induce aneuploidy by partial depletion of CENP-E in human primary fbroblasts (IMR90) and in near diploid tumor cells (HCT116). In contrast to IMR90 aneuploid cell number, which was drastically reduced and leaned towards the WT condition, HCT116 aneuploid cell numbers were slightly decreased at later time points. This euploidy restoration was accompanied by increased p14 ARF expression in IMR90 cells and followed ectopic p14 ARF re-expression in p14 ARF -null HCT116 cells. Collectively, our results suggest that hampering proliferation of aneuploid cells could be an additional role of the p14 ARF tumor suppressor. Keywords Aneuploidy · CENP-E · P14ARF · HCT116 cells · RNAi Introduction Accurate control of chromosome segregation during mitosis is a crucial step to prevent numerical and structural chromosomal changes leading to aneuploidy. Errors resulting in aneuploidy include improper kinetochore-microtubule attachments, cen- trosome amplifcation, chromosome cohesion defects and malfunctioning of the spindle assembly checkpoint (SAC) surveillance pathway (Gordon et al. 2012; Thompson et al. 2010). The SAC ensures chromosomal stability by preventing anaphase onset until all sister chromatids are properly attached by their kinetochores to the mitotic spindle. The depletion of some of its components weakens the SAC, resulting in defective mitoses which in turn will generate aneuploid cells (Thompson and Compton 2011). Aneuploidy has detrimental efects on both cultured cells and organisms of many species, including humans (Torres et al. 2008). Aneuploidy results in a dangerous genetic imbalance that has been shown to inhibit cell proliferation and to induce premature cellular senescence (Torres et al. 2007; Lentini et al. 2012). Despite these delete- rious consequences aneuploidy is a well-known hallmark of many solid tumors, conferring potentially “benefcial” charac- teristics to the developing cancer cells through the activation of oncogenes and repression of tumor suppressor genes (Bak- houm and Compton 2012). However, whether aneuploidy is a cause or consequence of cell transformation is still debated. Nevertheless, human aneuploidy syndromes, including Mosaic Variegate Aneuploidy (MVA), full constitutional trisomy 8 Communicated by Andres Aguilera. Lorena Veneziano and Viviana Barra contributed equally to this study. Electronic supplementary material The online version of this article (https://doi.org/10.1007/s00438-018-1495-5) contains supplementary material, which is available to authorized users. * Aldo Di Leonardo aldo.dileonardo@unipa.it 1 Department of Biological, Chemical and Pharmaceutical Sciences and Technologies, University of Palermo, Palermo, Italy 2 Present Address: Department of Genetic Stability and Oncogenesis, Institut Gustave Roussy, CNRS UMR8200, 94805 Villejuif, France 3 Centro di OncoBiologia Sperimentale (COBS), Palermo, Italy