BACKGROUND & AIMS: Caco-2 cells have been used extensively to elucidate events involved in intestinal cell proliferation and differentiation. Because individual isoforms of protein kinase C (PKC) and p21waf1, a cyclin-dependent kinase inhibitor, may regulate these processes, their role(s) on the growth and differentiation of Caco-2 cells were assessed. METHODS: Protein abundance and subcellular distribution of several PKC isoforms, as well as the expression of p21waf1, were examined in preconfluent and postconfluent cells. RESULTS: In cells at confluence (approximately 7 days postplating) and during their postconfluent phase (up to 20 days postplating), both total protein expression of PKC-alpha and its particulate distribution increased compared with their 3-day postplated counterparts. These findings were in agreement with those obtained by immunocytochemistry of PKC-alpha. In contrast, neither the total expression nor the subcellular distribution of PKC-betaI, -betaII, -delta, or -zeta changed significantly during these time periods. In addition, the expression of p21waf1, which can be induced by PKC-alpha, increased in postconfluent cells. CONCLUSIONS: PKC-alpha, but not other isoforms of PKC, may modulate the proliferation and differentiation of Caco-2 cells. This regulation appears to be mediated, at least in part, via a mechanism involving p21waf1.
BACKGROUND & AIMS: Caco-2 cells have been used extensively to elucidate events involved in intestinal cell proliferation and differentiation. Because individual isoforms of protein kinase C (PKC) and p21waf1, a cyclin-dependent kinase inhibitor, may regulate these processes, their role(s) on the growth and differentiation of Caco-2 cells were assessed. METHODS: Protein abundance and subcellular distribution of several PKC isoforms, as well as the expression of p21waf1, were examined in preconfluent and postconfluent cells. RESULTS: In cells at confluence (approximately 7 days postplating) and during their postconfluent phase (up to 20 days postplating), both total protein expression of PKC-alpha and its particulate distribution increased compared with their 3-day postplated counterparts. These findings were in agreement with those obtained by immunocytochemistry of PKC-alpha. In contrast, neither the total expression nor the subcellular distribution of PKC-betaI, -betaII, -delta, or -zeta changed significantly during these time periods. In addition, the expression of p21waf1, which can be induced by PKC-alpha, increased in postconfluent cells. CONCLUSIONS:PKC-alpha, but not other isoforms of PKC, may modulate the proliferation and differentiation of Caco-2 cells. This regulation appears to be mediated, at least in part, via a mechanism involving p21waf1.
Authors: Marybeth A Pysz; Olga V Leontieva; Nicholas W Bateman; Joshua M Uronis; Kathryn J Curry; David W Threadgill; Klaus-Peter Janssen; Sylvie Robine; Anna Velcich; Leonard H Augenlicht; Adrian R Black; Jennifer D Black Journal: Exp Cell Res Date: 2009-02-14 Impact factor: 3.905