PKCδ/midkine pathway drives hypoxia-induced proliferation and differentiation of human lung epithelial cells

Am J Physiol Cell Physiol. 2014 Apr 1;306(7):C648-58. doi: 10.1152/ajpcell.00351.2013. Epub 2014 Feb 5.


Epithelial cells are key players in the pathobiology of numerous hypoxia-induced lung diseases. The mechanisms mediating such hypoxic responses of epithelial cells are not well characterized. Earlier studies reported that hypoxia stimulates protein kinase C (PKC)δ activation in renal cancer cells and an increase in expression of a heparin-binding growth factor, midkine (MK), in lung alveolar epithelial cells. We reasoned that hypoxia might regulate MK levels via a PKCδ-dependent pathway and hypothesized that PKCδ-driven MK expression is required for hypoxia-induced lung epithelial cell proliferation and differentiation. Replication of human lung epithelial cells (A549) was significantly increased by chronic hypoxia (1% O2) and was dependent on expression of PKCδ. Hypoxia-induced proliferation of epithelial cells was accompanied by translocation of PKCδ from Golgi into the nuclei. Marked attenuation in MK protein levels by rottlerin, a pharmacological antagonist of PKC, and by small interfering RNA-targeting PKCδ, revealed that PKCδ is required for MK expression in both normoxic and hypoxic lung epithelial cells. Sequestering MK secreted into the culture media with a neutralizing antibody reduced hypoxia-induced proliferation demonstrating that an increase in MK release from cells is linked with epithelial cell division under hypoxia. In addition, recombinant MK accelerated transition of hypoxic epithelial cells to cells of mesenchymal phenotype characterized by elongated morphology and increased expression of mesenchymal markers, α-smooth muscle actin, and vimentin. We conclude that PKCδ/MK axis mediates hypoxic proliferation and differentiation of lung epithelial cells. Manipulation of PKCδ and MK activity in epithelial cells might be beneficial for the treatment of hypoxia-mediated lung diseases.

Keywords: PKCδ; differentiation; hypoxia; lung epithelial cells; midkine; proliferation.

Publication types

  • Research Support, N.I.H., Extramural

MeSH terms

  • Actins / metabolism
  • Active Transport, Cell Nucleus
  • Antibodies, Neutralizing / pharmacology
  • Biomarkers / metabolism
  • Cell Differentiation* / drug effects
  • Cell Hypoxia
  • Cell Line, Tumor
  • Cell Nucleus / enzymology
  • Cell Proliferation* / drug effects
  • Epithelial Cells / drug effects
  • Epithelial Cells / enzymology*
  • Epithelial Cells / pathology
  • Epithelial-Mesenchymal Transition
  • Golgi Apparatus / enzymology
  • Humans
  • Lung / drug effects
  • Lung / enzymology*
  • Lung / pathology
  • Midkine
  • Nerve Growth Factors / antagonists & inhibitors
  • Nerve Growth Factors / immunology
  • Nerve Growth Factors / metabolism*
  • Phenotype
  • Protein Kinase C-delta / antagonists & inhibitors
  • Protein Kinase C-delta / genetics
  • Protein Kinase C-delta / metabolism*
  • Protein Kinase Inhibitors / pharmacology
  • RNA Interference
  • Recombinant Proteins / metabolism
  • Signal Transduction
  • Time Factors
  • Transfection
  • Vimentin / metabolism


  • ACTA2 protein, human
  • Actins
  • Antibodies, Neutralizing
  • Biomarkers
  • MDK protein, human
  • Nerve Growth Factors
  • Protein Kinase Inhibitors
  • Recombinant Proteins
  • Vimentin
  • Midkine
  • PRKCD protein, human
  • Protein Kinase C-delta