title: Immortalization of MEF is characterized by the deregulation of specific miRNAs with potential tumor suppressor activity.
abstract: The life span (Hayflick limit) of primary mouse embryo fibroblasts (MEF) in culture is variable but it is still unclear if the escape of the Hayflick limit is also variable. To address this point MEF were expanded every fifteen days (6T15) instead of every three days (6T3) until they became immortal. With this protocol MEF lifespan was extended and immortalization accordingly delayed. By testing a panel of genes (p19ARF, p16, p21) and miRNAs (miR-20a, miR-21, miR-28, miR-290) related to primary MEF senescence, a switch of p21 from up to down regulation, the down regulation of specific miRNAs as well as a massive shift from diploidy to hyperdiploidy were observed in coincidence with the resumption of cell proliferation. Collectively, these data indicate that the inactivation of genes and miRNAs, important in controlling cell proliferation, might be determinant for the escape from the Hayflick limit. In support of this hypothesis was the finding that some of the down regulated miRNAs transfected in immortalized MEF inhibited cell proliferation thus displaying a tumor suppressor-like activity.
authors.affiliation: Laboratory of Gene and Molecular Therapy, Institute of Clinical Physiology, CNR, Pisa, Italy.
authors.forename: Milena
authors.initials: M
authors.lastname: Rizzo
authors.forename: Monica
authors.initials: M
authors.lastname: Evangelista
authors.forename: Marcella
authors.initials: M
authors.lastname: Simili
authors.forename: Laura
authors.initials: L
authors.lastname: Mariani
authors.forename: Letizia
authors.initials: L
authors.lastname: Pitto
authors.forename: Giuseppe
authors.initials: G
authors.lastname: Rainaldi
mesh_terms: D000818:Animals; D049109:Cell Proliferation; D002478:Cells, Cultured; D016922:Cellular Senescence / Q000502:physiology*; D005347:Fibroblasts / Q000166:cytology / Q000502:physiology*; D005786:Gene Expression Regulation; D016147:Genes, Tumor Suppressor; D051379:Mice; D035683:MicroRNAs / Q000235:genetics / Q000378:metabolism*
chemical_list: D035683:MicroRNAs