title: An antisense of protein kinase C-zeta inhibits proliferation of human airway smooth muscle cells.
abstract: We hypothesized that an atypical isoform of protein kinase (PK) C, PKC-zeta, 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-zeta and the wild-type PKC-zeta (Ad-CMV-PKC-zeta) 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-zeta protein, but not PKC-alpha or PKC-epsilon protein, in cells infected with the antisense when compared with levels in control cells. Thus, in HASM cells, PKC-zeta is involved in proliferation in response to PDGF, but not in response to FBS, for which alternate signal transduction pathways independent of PKC-zeta must exist.
authors.affiliation: Department of Pharmacology, University of Sydney, New South Wales, Australia.
authors.forename: S
authors.initials: S
authors.lastname: Carlin
authors.forename: P
authors.initials: P
authors.lastname: Poronnik
authors.forename: D I
authors.initials: DI
authors.lastname: Cook
authors.forename: L
authors.initials: L
authors.lastname: Carpenter
authors.forename: T J
authors.initials: TJ
authors.lastname: Biden
authors.forename: P R
authors.initials: PR
authors.lastname: Johnson
authors.forename: J L
authors.initials: JL
authors.lastname: Black
mesh_terms: D000256:Adenoviridae / Q000235:genetics; D000818:Animals; D002417:Cattle; D002455:Cell Division / Q000187:drug effects*; D002460:Cell Line; D049452:Green Fluorescent Proteins; D006801:Humans; D007527:Isoenzymes / Q000235:genetics*; D008164:Luminescent Proteins / Q000235:genetics; D009130:Muscle, Smooth / Q000166:cytology / Q000187:drug effects*; D016376:Oligonucleotides, Antisense / Q000494:pharmacology*; D011493:Protein Kinase C / Q000235:genetics*; D014132:Trachea / Q000166:cytology / Q000187:drug effects*; D015854:Up-Regulation
chemical_list: D007527:Isoenzymes; D008164:Luminescent Proteins; D016376:Oligonucleotides, Antisense; D049452:Green Fluorescent Proteins; C081975:protein kinase C eta; D011493:Protein Kinase C