title: PPAR-gamma expression in pituitary tumours and the functional activity of the glitazones: evidence that any anti-proliferative effect of the glitazones is independent of the PPAR-gamma receptor.
abstract: OBJECTIVE
  It has been reported that both normal pituitary and pituitary tumours express PPAR-gamma, a nuclear hormone receptor, the expression being more abundant in pituitary tumours, and that this is the basis for the reported antiproliferative effects of the thiazolidinedione, rosiglitazone, in animal models. However, the mechanisms for the responsivity to rosiglitazone have remained unclear.
  
  
  METHODS
  To investigate this further, 'real-time' PCR was used to assess PPAR-gamma mRNA expression, and Western blotting and immunohistochemistry to study its protein expression, in 46 human pituitary tumours and normal pituitary tissue. Cell proliferation of the GH3 pituitary cell line was assessed by [3H]-thymidine-incorporation after 48 h rosiglitazone and pioglitazone (10(-4) M- 10(-10) M) treatment alone, or rosiglitazone in combination with the PPAR-gamma antagonist GW9662.
  
  
  RESULTS
  PPAR-gamma mRNA and protein was found to be expressed in normal pituitary and was variably expressed in pituitary tumours, but were increased specifically in nonfunctioning pituitary adenomas. However, very little staining was observed with immunohistochemistry, with only occasional cell nuclei stained, and no difference was detectable between controls and tumours. Rosiglitazone at 10(-4) M and 10(-5) M concentrations inhibited cell proliferation (10(-4) M 14.0% +/- 1.5% and 10(-5) M 67% +/- 4%[mean +/- SEM]vs Control 100% +/- 3%, P < 0.0001) while lower concentrations showed no significant effect. Following withdrawal of rosiglitazone 10(-5) M, the cells fully recovered at a further 48 h, while lower doses showed a 'rebound' of stimulation. Pioglitazone was of similar potency to rosiglitazone in inhibiting proliferation. The PPAR-gamma antagonist did not show a significant reversal of the antiproliferative effect of rosiglitazone, and indeed suppressed proliferation on its own.
  
  
  CONCLUSIONS
  Our data suggest that the antiproliferative action of rosiglitazone is probably not via PPAR-gamma.
authors.affiliation: Department of Endocrinology, St. Bartholomew's and the Royal London School of Medicine and Dentistry, London, UK.
authors.forename: Michelle N
authors.initials: MN
authors.lastname: Emery
authors.forename: Chrysanthia
authors.initials: C
authors.lastname: Leontiou
authors.forename: Sarah E
authors.initials: SE
authors.lastname: Bonner
authors.forename: Chiara
authors.initials: C
authors.lastname: Merulli
authors.forename: Alexandra M
authors.initials: AM
authors.lastname: Nanzer
authors.forename: Madalina
authors.initials: M
authors.lastname: Musat
authors.forename: Malcolm
authors.initials: M
authors.lastname: Galloway
authors.forename: Michael
authors.initials: M
authors.lastname: Powell
authors.forename: Khash
authors.initials: K
authors.lastname: Nikookam
authors.forename: Márta
authors.initials: M
authors.lastname: Korbonits
authors.forename: Ashley B
authors.initials: AB
authors.lastname: Grossman
mesh_terms: D000236:Adenoma / Q000737:chemistry* / Q000473:pathology; D000328:Adult; D000368:Aged; D000369:Aged, 80 and over; D000818:Animals; D001483:Base Sequence; D016022:Case-Control Studies; D045744:Cell Line, Tumor; D049109:Cell Proliferation / Q000187:drug effects; D006801:Humans; D007150:Immunohistochemistry / Q000379:methods; D008875:Middle Aged; D008969:Molecular Sequence Data; D047495:PPAR gamma / Q000032:analysis* / Q000235:genetics; D047492:Peroxisome Proliferator-Activated Receptors / Q000378:metabolism*; D000077205:Pioglitazone; D010911:Pituitary Neoplasms / Q000737:chemistry* / Q000473:pathology; D051381:Rats; D020133:Reverse Transcriptase Polymerase Chain Reaction; D000077154:Rosiglitazone; D045162:Thiazolidinediones / Q000378:metabolism / Q000494:pharmacology*
chemical_list: D047495:PPAR gamma; D047492:Peroxisome Proliferator-Activated Receptors; D045162:Thiazolidinediones; D000077154:Rosiglitazone; D000077205:Pioglitazone