Butyrate-induced differentiation of Caco-2 cells is associated with apoptosis and early induction of p21Waf1/Cip1 and p27Kip1

Surgery. 1998 Aug;124(2):161-9; discussion 169-70.


Background: Intestinal mucosal turnover is a process of proliferation, differentiation, and apoptosis; the mechanisms remain largely undefined. The purpose of our study was to (1) assess the relationship between apoptosis and enterocyte differentiation and (2) determine whether the cell-cycle inhibitors, p21Waf1/Cip1 and p27Kip1, or the apoptosis inhibitors, Bcl-2 and Bcl-XL, may be involved.

Methods: Gut-derived Caco-2 cells were treated with sodium butyrate. Apoptosis was assessed by Hoechst stain, DNA laddering, and annexin V assay; differentiation was determined by alkaline phosphatase and sucrase activity. RNA and protein were analyzed for expression of p21Waf1/Cip1, p27Kip1, and members of the Bcl-2 family.

Results: Treatment of Caco-2 cells with sodium butyrate resulted in the concomitant induction of both differentiation (increased alkaline phosphatase and sucrase activity) and apoptosis. Increased levels of p21Waf1/Cip1 and p27Kip1 mRNA and protein were detected at 24 hours, occurring before apoptosis or differentiation; decreased mRNA levels of Bcl-2 and Bcl-XL were noted at 24 hours.

Conclusions: Differentiation and apoptosis occurred simultaneously in Caco-2 cells, suggesting that apoptosis may be linked to enterocyte differentiation. The induction of p21Waf1/Cip1 and p27Kip1 and the down-regulation of Bcl-2 and Bcl-XL further suggest a link between the cell-cycle mechanisms regulating enterocyte differentiation and apoptosis.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Alkaline Phosphatase / analysis
  • Alkaline Phosphatase / metabolism
  • Annexin A5 / analysis
  • Apoptosis / drug effects*
  • Butyrates / pharmacology*
  • Butyric Acid
  • Caco-2 Cells / cytology
  • Caco-2 Cells / drug effects
  • Caco-2 Cells / physiology
  • Cell Cycle Proteins*
  • Cell Differentiation / drug effects
  • Cyclin-Dependent Kinase Inhibitor p21
  • Cyclin-Dependent Kinase Inhibitor p27
  • Cyclins / genetics*
  • Enzyme Inhibitors / metabolism*
  • Flow Cytometry
  • Gene Expression Regulation, Neoplastic / drug effects
  • Genes, Tumor Suppressor / physiology
  • Histamine Antagonists / pharmacology*
  • Humans
  • Microtubule-Associated Proteins / genetics*
  • Microvilli / enzymology
  • Proto-Oncogene Proteins c-bcl-2 / genetics
  • Ribonucleases
  • Sucrase / analysis
  • Sucrase / metabolism
  • Tumor Suppressor Proteins*


  • Annexin A5
  • Butyrates
  • CDKN1A protein, human
  • Cell Cycle Proteins
  • Cyclin-Dependent Kinase Inhibitor p21
  • Cyclins
  • Enzyme Inhibitors
  • Histamine Antagonists
  • Microtubule-Associated Proteins
  • Proto-Oncogene Proteins c-bcl-2
  • Tumor Suppressor Proteins
  • Butyric Acid
  • Cyclin-Dependent Kinase Inhibitor p27
  • Ribonucleases
  • Alkaline Phosphatase
  • Sucrase