Am. J. Respir. Cell Mol. Biol., Volume 23, Number 4, October, 2000 555-559

An Antisense of Protein Kinase C- Inhibits Proliferation of Human Airway Smooth Muscle Cells

Stephen Carlin, Philip Poronnik, David I. Cook, Lee Carpenter, Trevor J. Biden, Peter R. A. Johnson, and Judith L. Black

Departments of Pharmacology and Physiology, University of Sydney; and the Garvan Institute, Sydney, New South Wales, Australia

We hypothesized that an atypical isoform of protein kinase (PK) C, PKC-, is essential for proliferation of human airway smooth muscle (HASM) cells in primary culture. Recombinant replication-deficient E1-deleted adenoviruses (100 plaque-forming units [pfu]/cell) expressing the antisense of PKC- and the wild-type PKC- (Ad-CMV-PKC-) were added to actively growing cells that were subsequently incubated for 48 h in platelet-derived growth factor (PDGF) 40 ng/mL or 10% fetal bovine serum (FBS). Expression of the antisense at a virus concentration of 100 pfu/cell produced a significant (n = 3, P < 0.05) decrease in the mean manual cell count in the presence of PDGF to 37 ± 5% relative to that in cells with no virus (100%), whereas in cells infected with virus containing no construct, this figure was 102 ± 13%. The increase in cell number in response to FBS, however, was not affected by the presence of the antisense. Corresponding values for cells in 10% FBS were 100 ± 22%, 85 ± 22%, and 122 ± 18%. Western blotting revealed decreased levels of PKC- protein, but not PKC- or PKC- protein, in cells infected with the antisense when compared with levels in control cells. Thus, in HASM cells, PKC- is involved in proliferation in response to PDGF, but not in response to FBS, for which alternate signal transduction pathways independent of PKC- must exist.

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