title: Differential proteome analysis of replicative senescence in rat embryo fibroblasts.
abstract: Normal somatic cells undergo a finite number of divisions and then cease dividing whereas cancer cells are able to proliferate indefinitely. To identify the underlying mechanisms that limit the mitotic potential, a two-dimensional differential proteome analysis of replicative senescence in serially passaged rat embryo fibroblasts was undertaken. Triplicate independent two-dimensional gels containing over 1200 spots each were run, curated, and analyzed. This revealed 49 spots whose expression was altered more than 2-fold. Of these, 42 spots yielded positive protein identification by mass spectrometry comprising a variety of cytoskeletal, heat shock, and metabolic proteins, as well as proteins involved in trafficking, differentiation, and protein synthesis, turnover, and modification. These included gelsolin, a candidate tumor suppressor for breast cancer, and alpha-glucosidase II, a member of the family of glucosidases that includes klotho; a defect in klotho expression in mice results in a syndrome that resembles human aging. Changes in expression of TUC-1, -2, -4, and -4 beta, members of the TUC family critical for neuronal differentiation, were also identified. Some of the identified changes were also shown to occur in two other models of senescence, premature senescence of REF52 cells and replicative senescence of mouse embryo fibroblasts. The majority of these candidate proteins were unrecognized previously in replicative senescence. They are now implicated in a new role.
authors.affiliation: Ludwig Institute for Cancer Research, Royal Free and University College School of Medicine, Courtauld Building, 91 Riding House Street, London W1W 7BS, United Kingdom.
authors.forename: Silvia
authors.initials: S
authors.lastname: Benvenuti
authors.forename: Rainer
authors.initials: R
authors.lastname: Cramer
authors.forename: Christopher C
authors.initials: CC
authors.lastname: Quinn
authors.forename: Jim
authors.initials: J
authors.lastname: Bruce
authors.forename: Marketa
authors.initials: M
authors.lastname: Zvelebil
authors.forename: Steven
authors.initials: S
authors.lastname: Corless
authors.forename: Jacquelyn
authors.initials: J
authors.lastname: Bond
authors.forename: Alice
authors.initials: A
authors.lastname: Yang
authors.forename: Susan
authors.initials: S
authors.lastname: Hockfield
authors.forename: Alma L
authors.initials: AL
authors.lastname: Burlingame
authors.forename: Michael D
authors.initials: MD
authors.lastname: Waterfield
authors.forename: Parmjit S
authors.initials: PS
authors.lastname: Jat
mesh_terms: D000818:Animals; D015153:Blotting, Western; D002460:Cell Line; D002478:Cells, Cultured; D016922:Cellular Senescence; D002851:Chromatography, High Pressure Liquid; D003599:Cytoskeleton / Q000378:metabolism; D015180:Electrophoresis, Gel, Two-Dimensional; D005347:Fibroblasts / Q000166:cytology* / Q000378:metabolism; D018260:Gelsolin / Q000096:biosynthesis; D005966:Glucuronidase; D000090265:Klotho Proteins; D008565:Membrane Proteins / Q000096:biosynthesis; D051381:Rats; D017207:Rats, Sprague-Dawley; D020133:Reverse Transcriptase Polymerase Chain Reaction; D021241:Spectrometry, Mass, Electrospray Ionization; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D015854:Up-Regulation; D000520:alpha-Glucosidases / Q000096:biosynthesis
chemical_list: D018260:Gelsolin; D008565:Membrane Proteins; D000520:alpha-Glucosidases; D005966:Glucuronidase; D000090265:Klotho Proteins