Differential regulation of Cl- transport proteins by PKC in Calu-3 cells.
Cl- transport proteins expressed in a Calu-3 airway epithelial cell line were differentiated by function and regulation by protein kinase C (PKC) isotypes. mRNA expression of Cl- transporters was semiquantitated by RT-PCR after transfection with a sense or antisense oligonucleotide to the PKC isotypes that modulate the activity of the cystic fibrosis transmembrane conductance regulator [CFTR (PKC-epsilon)] or of the Na/K/2Cl (NKCC1) cotransporter (PKC-delta). Expression of NKCC1 and CFTR mRNAs and proteins was independent of antisense oligonucleotide treatment. Transport function was measured in cell monolayers grown on a plastic surface or on filter inserts. With both culture methods, the antisense oligonucleotide to PKC-epsilon decreased the amount of PKC-epsilon and reduced cAMP-dependent activation of CFTR but not alpha(1)-adrenergic activation of NKCC1. The antisense oligonucleotide to PKC-delta did not affect CFTR function but did block alpha(1)-adrenergic activation of NKCC1 and reduce PKC-delta mass. These results provide the first evidence for mRNA and protein expression of NKCC1 in Calu-3 cells and establish the differential regulation of CFTR and NKCC1 function by specific PKC isotypes at a site distal to mRNA expression and translation in airway epithelial cells.
The Cystic Fibrosis Center, Department of Pediatrics, Case Western Reserve University, BRB, Rm. 824, 2109 Adelbert Rd., Cleveland, OH 44106-4948, USA. cxl7@po.cwru.edu
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D002352:Carrier Proteins / Q000235:genetics / Q000378:metabolism* / Q000502:physiology; D002712:Chlorides / Q000378:metabolism*; D019005:Cystic Fibrosis Transmembrane Conductance Regulator / Q000235:genetics / Q000502:physiology; D006801:Humans; D009841:Oligonucleotides / Q000494:pharmacology; D011493:Protein Kinase C / Q000502:physiology*; D012333:RNA, Messenger / Q000378:metabolism; D028021:Sodium-Potassium-Chloride Symporters; D014407:Tumor Cells, Cultured
C505032:CFTR protein, human; D002352:Carrier Proteins; D002712:Chlorides; D009841:Oligonucleotides; D012333:RNA, Messenger; D028021:Sodium-Potassium-Chloride Symporters; D019005:Cystic Fibrosis Transmembrane Conductance Regulator; D011493:Protein Kinase C