title: TSH/cAMP up-regulate sarco/endoplasmic reticulum Ca2+-ATPases expression and activity in PC Cl3 thyroid cells.
abstract: OBJECTIVE
  We recently reported that the sarco/endoplasmic reticulum Ca2+-ATPase (SERCA) 2b is the SERCA form preferentially expressed in rat thyroid. Moreover, SERCA2b expression dramatically decreases in virally transformed, highly tumorigenic, PC Cl3 thyroid cells. These results suggest that, in the thyroid, SERCA2b, in addition to its housekeeping role, is linked to differentiation and is a regulated gene. We therefore sought to study the effect of TSH, the main regulator of thyroid function, on SERCA2b expression and activity.
  
  
  METHODS
  PC Cl3 cells were hormone starved in low-serum medium and stimulated for long (48 h) or short (1, 2 and 4 h) times. SERCA2b expression and activity were evaluated by Northern and Western blots, Ca2+-ATPase activity and Ca2+ store content.
  
  
  RESULTS
  In PC Cl3 cells, SERCA2b mRNA and protein were induced twofold by a 48-h long treatment with TSH. Long-term elevation (48 h) of intracellular cAMP levels, by forskolin or 8-Br-cAMP, had similar effects on SERCA2b mRNA and protein. We also measured Ca2+-ATPase activity and Ca2+ store content. Both long (48 h) and short (0.5-1 h) treatments with TSH, forskolin or 8-Br-cAMP induced a marked increase of SERCA2b activity. This effect was completely abolished by H89, a specific inhibitor of cAMP-dependent protein kinase A (PKA). TSH and 8-Br-cAMP increased Ca2+ store content after both long (48 h) and short (1-2 h) treatments.
  
  
  CONCLUSIONS
  These data suggested that TSH/cAMP acts as an important regulator of both SERCA2b expression and activity in the thyroid system, through PKA activation.
authors.affiliation: Dipartimento di Biologia e Patologia Cellulare e Molecolare L. Califano, Via S. Pansini 5, 80131 Napoli, Italy.
authors.forename: Luca
authors.initials: L
authors.lastname: Ulianich
authors.forename: Agnese
authors.initials: A
authors.lastname: Secondo
authors.forename: Stefania
authors.initials: S
authors.lastname: De Micheli
authors.forename: Sonia
authors.initials: S
authors.lastname: Treglia
authors.forename: Francesco
authors.initials: F
authors.lastname: Pacifico
authors.forename: Domenico
authors.initials: D
authors.lastname: Liguoro
authors.forename: Fortunato
authors.initials: F
authors.lastname: Moscato
authors.forename: Santo
authors.initials: S
authors.lastname: Marsigliante
authors.forename: Lucio
authors.initials: L
authors.lastname: Annunziato
authors.forename: Silvestro
authors.initials: S
authors.lastname: Formisano
authors.forename: Eduardo
authors.initials: E
authors.lastname: Consiglio
authors.forename: Bruno
authors.initials: B
authors.lastname: Di Jeso
mesh_terms: D015124:8-Bromo Cyclic Adenosine Monophosphate / Q000494:pharmacology; D000818:Animals; D000252:Calcium-Transporting ATPases / Q000235:genetics* / Q000378:metabolism; D002460:Cell Line; D005576:Colforsin / Q000494:pharmacology; D000242:Cyclic AMP / Q000494:pharmacology*; D017868:Cyclic AMP-Dependent Protein Kinases / Q000378:metabolism; D004789:Enzyme Activation; D005786:Gene Expression Regulation / Q000187:drug effects*; D012333:RNA, Messenger / Q000032:analysis; D051381:Rats; D020133:Reverse Transcriptase Polymerase Chain Reaction; D053498:Sarcoplasmic Reticulum Calcium-Transporting ATPases; D013961:Thyroid Gland / Q000201:enzymology*; D013972:Thyrotropin / Q000494:pharmacology*
chemical_list: D012333:RNA, Messenger; D005576:Colforsin; D015124:8-Bromo Cyclic Adenosine Monophosphate; D013972:Thyrotropin; D000242:Cyclic AMP; D017868:Cyclic AMP-Dependent Protein Kinases; D053498:Sarcoplasmic Reticulum Calcium-Transporting ATPases; D000252:Calcium-Transporting ATPases