Distinct functions of protein kinase Calpha and protein kinase Cbeta during retinoic acid-induced differentiation of F9 cells.
As F9 embryonal carcinoma cells differentiate into parietal endoderm-like cells, expression of conventional protein kinase C (PKC) changes. Undifferentiated stem cells express PKCbeta but not PKCalpha, whereas differentiated parietal endoderm cells express PKCalpha but not PKCbeta. To determine whether changes in PKCalpha and/or PKCbeta expression control retinoic acid (RA)- and dibutyryl cyclic AMP-induced F9 cell differentiation, we established cell lines stably expressing PKCalpha, PKCbeta, antisense PKCalpha, or antisense PKCbeta RNAs. Constitutive expression of PKCalpha or inhibition of PKCbeta expression in F9 stem cells enhanced RA induced differentiation, both by increasing total expression and accelerating RA-induced expression of laminins A, B1, B2, and type IV collagen. In addition, expressing PKCbeta in a parietal endoderm cell line caused these cells to retrodifferentiate into stem cells. Based on these results, we conclude that PKCbeta and PKCalpha are key targets for RA-regulated gene expression, that PKCalpha plays an important, active role in inducing and maintaining the parietal endoderm phenotype, and that PKCbeta activity is incompatible with maintaining the differentiated state of these cells.
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Institute of Human Nutrition, Columbia University, New York, New York 10032, USA.
Klein
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MG
Talmage
D A
DA
D000818:Animals; D003994:Bucladesine / Q000494:pharmacology; D002454:Cell Differentiation; D002460:Cell Line; D003094:Collagen / Q000235:genetics; D016373:DNA, Antisense; D054278:Embryonal Carcinoma Stem Cells; D004707:Endoderm / Q000166:cytology* / Q000201:enzymology; D005786:Gene Expression Regulation; D016762:Genes, fos; D007527:Isoenzymes / Q000235:genetics / Q000378:metabolism*; D051379:Mice; D014411:Neoplastic Stem Cells / Q000166:cytology* / Q000201:enzymology; D010641:Phenotype; D011493:Protein Kinase C / Q000235:genetics / Q000378:metabolism*; D064546:Protein Kinase C beta; D051571:Protein Kinase C-alpha; D015398:Signal Transduction; D014162:Transfection; D014212:Tretinoin / Q000494:pharmacology*; D014407:Tumor Cells, Cultured
D016373:DNA, Antisense; D007527:Isoenzymes; D014212:Tretinoin; D003994:Bucladesine; D003094:Collagen; C496630:Prkca protein, mouse; D011493:Protein Kinase C; D064546:Protein Kinase C beta; D051571:Protein Kinase C-alpha