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Am J Physiol Cell Physiol 277: C310-C319, 1999;
0363-6143/99 $5.00
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Vol. 277, Issue 2, C310-C319, August 1999

Antagonism of CD95 signaling blocks butyrate induction of apoptosis in young adult mouse colonic cells

Yang-Yi Fan1, Jianhu Zhang1, Rola Barhoumi2,3, Robert C. Burghardt2,3, Nancy D. Turner1, Joanne R. Lupton1,2, and Robert S. Chapkin1,2

1 Molecular and Cell Biology Group, Faculty of Nutrition, 2 Department of Veterinary Anatomy and Public Health, and 3 Image Analysis Laboratory, Texas A&M University, College Station, Texas 77843

There is great interest in utilizing butyrate as a chemopreventive agent for colon tumorigenesis because of its ability to promote apoptosis in colon tumor cell lines. Because CD95 (APO-1/Fas) transduces signals resulting in apoptosis, we tested the hypothesis that butyrate-dependent colonocyte apoptosis is mediated by this death receptor. Butyrate (1 mM) exposure for 24 h upregulated expression of Fas and its ligand in young adult mouse colon (YAMC) cells. To delineate the proapoptotic effect of butyrate and to avoid the confounding effects of detachment from the extracellular matrix, adherent cell apoptosis was monitored as loss of plasma membrane asymmetry and dissipation of mitochondrial membrane potential (Delta Psi mt) by laser cytometry. Soluble Fas receptor protein (Fas:Fc chimera) and caspase inhibitors (z-VAD-fmk and z-IETD-fmk) blocked butyrate induction of apoptosis. Treatment with Fas agonistic antibody (clone Jo-2) significantly induced cell death, indicating that Fas in colonocytes is functional. In addition, butyrate promoted apoptosis by inducing loss of Delta Psi mt and phospholipid asymmetry of the plasma membrane after 12 and 24 h of exposure, respectively, before cell detachment. Therefore, Fas receptor-dependent signal transduction is involved in butyrate induction of apoptosis in colonocytes.

anoikis; plasma membrane asymmetry; mitochondrial membrane potential; colon cancer; Fas; APO-1


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