Cellular Model of p21-Induced Senescence

Methods Mol Biol. 2017:1534:31-39. doi: 10.1007/978-1-4939-6670-7_3.

Abstract

Cellular senescence is a unique process of normal physiology, from embryonic development to aging, also known for its association with a broad range of pathological conditions. Therefore a reliable model of cellular senescence remains an indispensable tool for the investigation of senescence-associated changes and human disease. Here we describe a model of HT1080 fibrosarcoma cells with an inducible senescence phenotype. These cells are equipped with the lac repressor and exogenous p21 under the control of a lac repressor regulated promoter. The senescent phenotype is induced in these cells by isopropyl-β-D-thiogalactopyranoside (IPTG)-inducible expression of senescence-associated cell cycle inhibitor p21Waf1/Cip1/Sdi1.

Keywords: HT1080-p21-9; IPTG; Lac-repressor; Senescence; p21Waf1/Cip1/Sdi1.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Biomarkers
  • Cell Cycle / genetics
  • Cell Line
  • Cellular Senescence / physiology*
  • Cyclin-Dependent Kinase Inhibitor p21 / genetics*
  • Cyclin-Dependent Kinase Inhibitor p21 / metabolism*
  • Flow Cytometry
  • Gene Expression
  • Gene Knock-In Techniques
  • Humans
  • Phenotype

Substances

  • Biomarkers
  • Cyclin-Dependent Kinase Inhibitor p21