Protein kinase C-alpha regulates proliferation but not apoptosis in rat coronary vascular smooth muscle cells.
In a previous study (Am. J. Pathol. 1994, 145: 1265-1270) we found rat coronary vascular smooth muscle cell (SMC) proliferation and apoptosis to be regulated by protein kinase C (PKC). In the present study we analysed whether selective depletion of alpha isozyme of PKC would affect SMC proliferation and/or apoptosis. First, using Western blot technique, it was determined that the rat SMC express alpha, delta, epsilon and zeta isozymes of PKC. The selective depletion of PKC-alpha in SMC was achieved by exposing cells to antisense oligodeoxynucleotide to mRNA for PKC-alpha (AS-PKC-alpha). The effect of AS-PKC-alpha on SMC proliferation was analysed by measurement of 3H-thymidine incorporation. The results indicated that a single dose of AS-PKC-alpha at a concentration of 10-100microM caused long-lasting (for at least 4 days) inhibition (up to 55%) of 3H-thymidine incorporation by SMC. This observation indirectly demonstrates that PKC-alpha regulates SMC proliferation. However, it was not possible to induce a significant level of apoptosis in SMC exposed even to the highest dose of AS-PKC-alpha. These data, in conjunction with the previously shown induction of apoptosis in SMC by calphostin C, suggests that another isozyme of PKC is likely to be involved in regulation of SMC apoptosis. Finally, we observed that induction of apoptosis via PKC-dependent mechanism is prevented by supplementing the culture medium with serum. This shows striking similarity with the regulation of apoptosis by the c-myc-dependent pathway. In conclusion, PKC-alpha joins the group of proteins such as c-myc, proliferating-cell nuclear antigen and cdc2 kinase which may be therapeutical targets, for antisense oligodeoxynucleotides, in order to prevent SMC hyperplasia.
Leszczynski
D
D
Department of Research, Finnish Centre for Radiation and Nuclear Safety, Helsinki, Finland.
Joenväärä
S
S
Foegh
M L
ML
D000818:Animals; D017209:Apoptosis; D001483:Base Sequence; D002455:Cell Division; D002478:Cells, Cultured; D003331:Coronary Vessels / Q000166:cytology* / Q000201:enzymology; D066298:In Vitro Techniques; D007527:Isoenzymes / Q000378:metabolism*; D008969:Molecular Sequence Data; D009131:Muscle, Smooth, Vascular / Q000166:cytology* / Q000201:enzymology; D009838:Oligodeoxyribonucleotides; D011493:Protein Kinase C / Q000378:metabolism*; D051571:Protein Kinase C-alpha; D051381:Rats
D007527:Isoenzymes; D009838:Oligodeoxyribonucleotides; D011493:Protein Kinase C; D051571:Protein Kinase C-alpha