Lack of protein kinase C (PKC)-beta and low PKC-alpha, -delta, -epsilon, and -zeta isozyme levels in proliferating human melanoma cells.
Protein kinase C (PKC), a calcium and phospholipid-dependent kinase, has been implicated in carcinogenesis of melanocytic cells. However, its role in melanoma cell growth remains controversial. We therefore investigated the growth dependence of PKC isozyme expression in human normal melanocytes and melanoma cells. Logarithmic and stationary growth phases in culture were clearly distinguished by nuclear cell staining with the proliferation marker Ki-67. PKC-beta I and -beta II were expressed exclusively in normal melanocytes but not in melanoma cells, whereas PKC-gamma was not found in any of the cultures studied. Low PKC-delta, -epsilon and -zeta mRNA levels were detected by RT-PCR in proliferating melanoma cells and higher in confluent non-proliferating cells, whereas levels of PKC-alpha mRNA remained rather stable. Subcellular fractionation and immunoblotting revealed accordingly low expression of PKC-alpha, -delta, -epsilon, and -zeta in the logarithmic growth phase of melanoma cells, with subsequent increase of expression and of membrane association in the stationary phase. Only weak differences were detected between the growth phases in normal melanocytes for the respective PKC isozymes, except for membrane-associated PKC-beta I and -beta II which were clearly elevated in confluent melanocyte cultures. These data suggest that certain PKC isozymes are involved in the intracellular signalling that regulates melanoma cell proliferation, and may function as suppressors of tumour cell growth.
Krasagakis
Konstantin
K
Department of Dematology, University Medical Center Bejamin Franklin, Free University of Berlin, Germany. krasagak@med.uoc.gr
Fimmel
Sabine
S
Genten
Daniela
D
Eberle
Jürgen
J
Quas
Petra
P
Ziegler
Wolfgang
W
Haller
Hermann
H
Orfanos
Constantin E
CE
D015153:Blotting, Western; D002455:Cell Division / Q000502:physiology*; D002478:Cells, Cultured; D003600:Cytosol; D005786:Gene Expression Regulation / Q000502:physiology; D006801:Humans; D007527:Isoenzymes / Q000235:genetics / Q000378:metabolism*; D019394:Ki-67 Antigen / Q000378:metabolism; D008544:Melanocytes / Q000187:drug effects / Q000201:enzymology*; D008545:Melanoma / Q000201:enzymology* / Q000473:pathology; D011493:Protein Kinase C / Q000235:genetics / Q000378:metabolism*; D064546:Protein Kinase C beta; D012313:RNA / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction
D007527:Isoenzymes; D019394:Ki-67 Antigen; D012313:RNA; D011493:Protein Kinase C; D064546:Protein Kinase C beta