Evidence for LHRH-receptor expression in human airway epithelial (Calu-3) cells and its role in the transport of an LHRH agonist

Pharm Res. 2004 Jun;21(6):1034-46. doi: 10.1023/b:pham.0000029294.70707.74.


Purpose: To determine whether LHRH-receptor is expressed in Calu-3, a human bronchial epithelial cell line, and to further determine whether this receptor plays a role in the transport of deslorelin, an LHRH agonist.

Methods: Using cultured monolayers of Calu-3 grown at air-interface, the presence and localization of LHRH-receptors in Calu-3 cells was determined using immunochemical methods. To determine the mechanisms of deslorelin transport, the directionality [apical-basolateral (A-B) and basolateral-apical (B-A)] of deslorelin transport across Calu-3 monolayers and the effects of temperature (37 degrees C and 4 degrees C) and an energy depletor (2,4-dinitrophenol) were investigated. To determine the role of LHRH-receptor in deslorelin transport across Calu-3 monolayers, the influence of an LHRH-receptor antisense oligonucleotide on the LHRH-receptor expression and deslorelin transport was studied. Also, the effect of a competing LHRH agonist, buserelin, on deslorelin transport was determined.

Results: Immunofluorescence studies indicated the predominance of LHRH-receptor in Calu-3 cells at the apical and lateral surfaces. Western blot and RT-PCR studies further confirmed the expression of LHRH-receptor in Calu-3 cells. Deslorelin transport across Calu-3 monolayers was vectorial, with the cumulative A-B transport (1.79 +/- 0.29%) at the end of 240 min being higher than the B-A transport (0.34 +/- 0.11%). Low temperature as well as 2,4-dinitrophenol abolished this directionality. LHRH-receptor antisense oligonucleotide decreased the receptor expression at the mRNA and protein level and reduced the A-B deslorelin transport by 55 +/- 4%, without affecting the B-A transport, suggesting a role for LHRH-receptor in the vectorial transport of deslorelin. In addition, buserelin reduced the A-B deslorelin transport by 56 +/- 5% without affecting the B-A transport.

Conclusions: Taken together, our results provide evidence that deslorelin is transported across the respiratory epithelium via the LHRH-receptor.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • 2,4-Dinitrophenol / pharmacology
  • Animals
  • Biological Transport
  • Blotting, Western / methods
  • Buserelin / pharmacokinetics*
  • Buserelin / pharmacology
  • Cattle
  • Cell Line
  • Electric Impedance
  • Epithelial Cells / cytology*
  • Epithelial Cells / physiology
  • Fluorescent Antibody Technique / methods
  • Gene Expression / genetics*
  • Humans
  • Oligoribonucleotides, Antisense / pharmacology
  • RNA, Messenger
  • Rats
  • Receptors, LHRH / antagonists & inhibitors
  • Receptors, LHRH / genetics*
  • Receptors, LHRH / immunology
  • Respiratory System / cytology*
  • Respiratory System / metabolism
  • Reverse Transcriptase Polymerase Chain Reaction
  • Temperature
  • Time Factors
  • Triptorelin Pamoate / analogs & derivatives*
  • Triptorelin Pamoate / pharmacokinetics*
  • Triptorelin Pamoate / pharmacology


  • Oligoribonucleotides, Antisense
  • RNA, Messenger
  • Receptors, LHRH
  • Triptorelin Pamoate
  • Buserelin
  • 2,4-Dinitrophenol
  • deslorelin