Protein kinase C isoform alpha overexpression in C6 glioma cells and its role in cell proliferation.
Previous studies from this laboratory have demonstrated that protein kinase C (PKC) enzyme activity is highly correlated with the proliferation rate of glioma cells, and that glioma cells of both human and rat origin have very high PKC enzyme activity when compared to non-malignant glia including astrocytes, the antecedents of most gliomas. In the present study, by contrasting the rat C6 glioma cells with non-malignant rat astrocytes, we have sought to determine whether the high PKC enzyme activity of glioma cells was due to the overexpression of a specific isoform of PKC. By Western blot analyses, both C6 glioma cells and astrocytes were found to express PKC alpha, beta, delta, epsilon and zeta, but not gamma. Enzyme activity measurements revealed that the elevated PKC activity of glioma cells compared to glia was calcium-dependent, thereby implicating abnormal activity of the alpha or beta isoforms. On Western blots, when compared to astrocytes, glioma cells were determined to overexpress PKC alpha but not beta. An antisense oligonucleotide to PKC alpha, directed at the site of initiation of translation, inhibited the proliferation rate of glioma cells when compared to cells treated with control oligonucleotides; PKC enzyme activity and PKC alpha protein expression were significantly reduced by the antisense treatment. These results suggest that the high PKC enzyme activity of glioma cells, and its correspondence with proliferation rate, is the result of overexpression of isozyme alpha. Targetting PKC alpha in glioma cells may provide a refinement of therapy of glioma patients, some of which are already showing clinical stabilization when treated with drugs with PKC-inhibitory effects.
Baltuch
G H
GH
Department of Neurology and Neurosurgery, Montreal Neurological Institute and McGill University, Quebec, Canada.
Dooley
N P
NP
Rostworowski
K M
KM
Villemure
J G
JG
Yong
V W
VW
D000818:Animals; D001253:Astrocytes / Q000201:enzymology*; D001483:Base Sequence; D001932:Brain Neoplasms / Q000201:enzymology* / Q000473:pathology; D005910:Glioma / Q000201:enzymology* / Q000473:pathology; D006801:Humans; D007527:Isoenzymes / Q000096:biosynthesis*; D008969:Molecular Sequence Data; D016376:Oligonucleotides, Antisense / Q000494:pharmacology; D011493:Protein Kinase C / Q000096:biosynthesis*; D051571:Protein Kinase C-alpha; D051381:Rats; D014407:Tumor Cells, Cultured
D007527:Isoenzymes; D016376:Oligonucleotides, Antisense; C496628:PRKCA protein, human; D011493:Protein Kinase C; D051571:Protein Kinase C-alpha