Bile acid deoxycholate induces differential subcellular localisation of the PKC isoenzymes beta 1, epsilon and delta in colonic epithelial cells in a sodium butyrate insensitive manner.
Elevated levels of bile acids have been implicated in the abnormal morphogenesis of the colonic epithelium thus contributing to colorectal cancer (CRC). Alternatively sodium butyrate (NaB) produced by anaerobic fermentation of dietary fibre is regarded as being protective against colon cancer. Bile acids such as deoxycholic acid (DCA) are thought to mediate some of their actions by differentially activating protein kinase C (PKC). We examined the effects of DCA on the subcellular localisation of PKC-beta(1), -epsilon and -delta and whether these responses could be modulated by NaB. HCT116 cells endogenously express PKC-epsilon and -delta but not PKC-beta. DCA treatment results in endogenous PKC-epsilon translocation but not PKC-delta after 1 hr. To study the subcellular localisation of PKC isoforms in response to DCA in real time, PKC-beta(1), PKC-epsilon and PKC-delta functionally intact green fluorescent protein (GFP) fusion constructs were used. Stimulation with 300 microM DCA induces rapid translocation of PKC-beta(1)-GFP and PKC-epsilon-GFP but not PKC-delta-GFP from the cytosol to the plasma membrane in 15 min. Interestingly, pretreatment with 4mM NaB does not modify the response of the PKC isoenzymes to DCA as PKC-beta(1)-GFP and PKC-epsilon-GFP translocates to the plasma membrane in 15 min whereas PKC-delta-GFP localisation remains unaltered. Immunofluorescence shows that PKC-beta(1)-GFP and PKC-epsilon-GFP cells treated with DCA colocalise with the cytoskeletal elements actin and tubulin adjacent to the plasma membrane. Our findings demonstrate that the differential activation of the PKC isoenzymes by DCA may be of critical importance for the functional responses of colonic epithelial cells. Supplementary material for this article can be found on the International Journal of Cancer website at http://www.interscience.wiley.com/jpages/0020-7136/suppmat/index.html.
Looby
Eileen
E
Department of Clinical Medicine, Trinity College and Dublin Molecular Medicine Centre, Dublin, Ireland.
Long
Aideen
A
Kelleher
Dermot
D
Volkov
Yuri
Y
D002087:Butyrates / Q000494:pharmacology*; D018929:Cell Culture Techniques; D003106:Colon / Q000166:cytology* / Q000473:pathology; D015179:Colorectal Neoplasms / Q000209:etiology* / Q000517:prevention & control*; D003599:Cytoskeleton; D003840:Deoxycholic Acid / Q000494:pharmacology*; D003902:Detergents / Q000494:pharmacology*; D004789:Enzyme Activation; D004848:Epithelium; D006801:Humans; D058610:Isobutyrates; D007527:Isoenzymes; D011493:Protein Kinase C / Q000493:pharmacokinetics* / Q000494:pharmacology*
D002087:Butyrates; D003902:Detergents; D058610:Isobutyrates; D007527:Isoenzymes; D003840:Deoxycholic Acid; C020380:isobutyric acid; D011493:Protein Kinase C