Cloning and identification of genes that associate with mammalian replicative senescence

Exp Cell Res. 1998 Apr 10;240(1):66-74. doi: 10.1006/excr.1998.3948.

Abstract

Cellular senescence and limited proliferative capacity of normal diploid cells has a dominant phenotype over immortality of cancerous cells, suggesting its regulation by the expression of a set of genes. In order to isolate the genes that associate with senescence, we have employed a clonal system of conditional SV40 T antigen rat embryo fibroblast cell lines which undergo senescence upon T antigen inactivation. Construction of cDNA libraries from two conditional cell lines and application of differential screening and subtractive hybridization techniques have resulted in the cloning of eight senescence-induced genes (SGP-2/Apo J, alpha 1-procollagen, osteonectin, fibronectin, SM22, cytochrome C oxidase, GTP-alpha, and a novel gene) and a senescence-repressed gene (FRS-2). Three of these genes encode for extracellular matrix proteins, others are involved in the calcium-dependent signal transduction pathways, while the SGP-2/Apo J gene may have a cellular protective function. RNA analysis has shown that the senescence-associated genes are overexpressed in both normal rat embryonic fibroblasts and human osteoblasts cell cultures undergoing aging in vitro. In comparison, the expression of these genes in a rat fibroblast immortalized cell line (208F cells) was down-regulated after both its partial and its full transformation by ras oncogenes. Thus, cloning of senescence-associated genes opens up new ways to elucidate and/or to modulate aging and cancer.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Aging / genetics*
  • Animals
  • Cell Line, Transformed / cytology
  • Cell Line, Transformed / physiology
  • Cellular Senescence / genetics
  • Cloning, Molecular
  • DNA Replication / genetics*
  • DNA, Complementary / analysis
  • Fibroblasts / cytology
  • Fibroblasts / physiology
  • Gene Expression Regulation, Neoplastic / genetics*
  • Gene Library
  • Glycoproteins / genetics
  • Humans
  • Mammals
  • Osteoblasts / cytology*
  • Osteoblasts / physiology
  • RNA, Messenger / genetics
  • Rats

Substances

  • DNA, Complementary
  • Glycoproteins
  • RNA, Messenger