Decreased expression of the type I isozyme of cAMP-dependent protein kinase in tumor cell lines of lung epithelial origin.
A spontaneous transformant derived from a mouse lung epithelial cell line exhibited decreased cAMP-dependent protein kinase (PKA) activity. DEAE column chromatography demonstrated that this was caused by specific loss of the type I PKA isozyme (PKA I). Western immunoblot analysis indicated that indeed several mouse lung tumor-derived cell lines and spontaneous transformants of immortalized, nontumorigenic lung cell lines contained less PKA I regulatory subunit (RI) protein than normal cell lines. PKA II regulatory subunit protein differed only slightly among cell lines and showed no conspicuous trend between normal and neoplastic cells. The decrease in RI was apparently concomitant with decreased catalytic (C) subunit levels in neoplastic cells since no free catalytic subunit activity was detected by DEAE chromatography. Northern blot analysis using RI alpha and C alpha cDNA probes showed that the levels of RI alpha and C alpha mRNAs paralleled their intracellular protein concentrations; neoplastic cell lines contained significantly less RI alpha and C alpha mRNAs than the normal cell line. The decreased expression of both RI and C subunits therefore results in a net decrease of PKA I in neoplastic lung cells, an isozymic difference which may account for the differential effects of cAMP analogs on cell growth and differentiation in normal and neoplastic cells.
Lange-Carter
C A
CA
Molecular and Environmental Toxicology Program, School of Pharmacy, University of Colorado, Boulder 80309.
Fossli
T
T
Jahnsen
T
T
Malkinson
A M
AM
D000249:Adenosine Monophosphate / Q000031:analogs & derivatives / Q000378:metabolism; D000345:Affinity Labels; D000818:Animals; D001386:Azides / Q000378:metabolism; D015153:Blotting, Western; D002460:Cell Line; D002848:Chromatography, DEAE-Cellulose; D000242:Cyclic AMP / Q000494:pharmacology*; D015342:DNA Probes; D007527:Isoenzymes / Q000235:genetics* / Q000378:metabolism; D008175:Lung Neoplasms / Q000201:enzymology*; D046911:Macromolecular Substances; D051379:Mice; D010446:Peptide Fragments / Q000378:metabolism; D011494:Protein Kinases / Q000235:genetics* / Q000378:metabolism; D012333:RNA, Messenger / Q000378:metabolism; D015398:Signal Transduction; D014407:Tumor Cells, Cultured
D000345:Affinity Labels; D001386:Azides; D015342:DNA Probes; D007527:Isoenzymes; D046911:Macromolecular Substances; D010446:Peptide Fragments; D012333:RNA, Messenger; D000249:Adenosine Monophosphate; C021291:8-azidoadenosine 5'-monophosphate; D000242:Cyclic AMP; D011494:Protein Kinases