An antisense of protein kinase C-zeta inhibits proliferation of human airway smooth muscle cells

Am J Respir Cell Mol Biol. 2000 Oct;23(4):555-9. doi: 10.1165/ajrcmb.23.4.4197.

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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adenoviridae / genetics
  • Animals
  • Cattle
  • Cell Division / drug effects*
  • Cell Line
  • Green Fluorescent Proteins
  • Humans
  • Isoenzymes / genetics*
  • Luminescent Proteins / genetics
  • Muscle, Smooth / cytology
  • Muscle, Smooth / drug effects*
  • Oligonucleotides, Antisense / pharmacology*
  • Protein Kinase C / genetics*
  • Trachea / cytology
  • Trachea / drug effects*
  • Up-Regulation

Substances

  • Isoenzymes
  • Luminescent Proteins
  • Oligonucleotides, Antisense
  • Green Fluorescent Proteins
  • protein kinase C eta
  • Protein Kinase C