Functional inactivation of pRB results in aneuploid mammalian cells after release from a mitotic block.
The widespread chromosome instability observed in tumors and in early stage carcinomas suggests that aneuploidy could be a prerequisite for cellular transformation and tumor initiation. Defects in tumor suppressors and genes that are part of mitotic checkpoints are likely candidates for the aneuploid phenotype. By using flow cytometric, cytogenetic, and immunocytochemistry techniques we investigated whether pRB deficiency could drive perpetual aneuploidy in normal human and mouse fibroblasts after mitotic checkpoint challenge by microtubule-destabilizing drugs. Both mouse and human pRB-deficient primary fibroblasts resulted, upon release from a mitotic block, in proliferating aneuploid cells possessing supernumerary centrosomes. Aneuploid pRB-deficient cells show an elevated variation in chromosome numbers among cells of the same clone. In addition, these cells acquired the capability to grow in an anchorage-independent way at the same extent as tumor cells did suggesting aneuploidy as an initial mutational step in cell transformation. Normal Mouse Embryonic Fibroblasts (MEFs) harboring LoxP sites flanking exon 19 of the Rb gene arrested in G2/M with duplicated centrosomes after colcemid treatment. However, these cells escaped the arrest and became aneuploid upon pRB ablation by CRE recombinase, suggesting pRB as a major component of a checkpoint that controls cellular ploidy.
Lentini
Laura
L
Department of Cell and Developmental Biology A. Monroy, University of Palermo, viale delle Scienze 90128, Palermo, Italy.
Pipitone
Loredana
L
Di Leonardo
Aldo
A
D000782:Aneuploidy; D000818:Animals; D000972:Antineoplastic Agents, Phytogenic / Q000494:pharmacology; D002453:Cell Cycle / Q000187:drug effects; D002471:Cell Transformation, Neoplastic; D002478:Cells, Cultured; D002873:Chromosome Fragility; D003114:Colony-Forming Units Assay; D004261:DNA Replication / Q000187:drug effects*; D003703:Demecolcine / Q000494:pharmacology; D005260:Female; D005347:Fibroblasts / Q000166:cytology / Q000187:drug effects*; D005434:Flow Cytometry; D016161:Genes, Retinoblastoma / Q000502:physiology*; D006801:Humans; D017404:In Situ Hybridization, Fluorescence; D051379:Mice / Q000196:embryology; D008856:Microscopy, Fluorescence; D008938:Mitosis / Q000235:genetics; D016160:Retinoblastoma Protein / Q000172:deficiency; D008941:Spindle Apparatus / Q000235:genetics
D000972:Antineoplastic Agents, Phytogenic; D016160:Retinoblastoma Protein; D003703:Demecolcine