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.
Liedtke
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CM
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
Cody
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Cole
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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