Lovastatin decreases prolactin and growth hormone gene expression in GH4C1 cells through a cAMP dependent mechanism.
The heterotrimeric G protein Gs couples several surface ligand receptors to cAMP production, as well as to both growth hormone (GH) and prolactin (PRL) gene expression in pituitary and GH cells. It has been shown that constitutively active alpha s stimulates transient expression of both PRL- and GH- chloramphenicol acetyl transferase (CAT) constructions, which indicates that both the PRL and GH promoter regions are under the influence of signal pathways mediated by alpha s. We have previously shown that the cholesterol lowering drug lovastatin decreases both the amount of G alpha s subunit in the membrane and the adenylyl cyclase activity in GH4C1 cells. Thus, we tried to verify whether that decrease in alpha s levels could affect PRL and GH secretion, as well as the expression of PRL- and GH-CAT constructions. Since the regulation of these two genes is dependent on the pituitary specific transcription factor Pit-1, the effect of lovastatin on the expression of Pit-1-CAT constructions was also studied. Our results show that lovastatin decreased the basal expression of these three cAMP-responsive genes in GH4C1 cells, being partially reversed by the addition of mevalonate to the culture medium. This effect of lovastatin on the promoter activities of the transfected constructions was also observed in PRL and GH secretion to the medium, suggesting that this drug produces similar changes in the endogenous promoters of both hormones. Moreover, the presence of lovastatin did not prevent the response to the cAMP activator forskolin, indicating that the main effect of this drug could be exerted through upstream adenylyl cyclase. In conclusion, our data indicate that lovastatin decreases the basal expression of Pit-1 and consequently of both GH and PRL genes through a mechanism probably mediated by the decrease of G alpha s levels in the cell membrane. Taken together, these results suggest that the activity of membrane heterotrimeric G proteins regulates the basal transcription of specific cellular genes in GH4C1 cells. Moreover the effects of lovastatin may be taken into account in the study of constitutively endocrine disorders associated with an increased secretion of either PRL or GH.
Lasa
M
M
Departamento de Bioquímica y Biología Molecular, Universidad de Alcalá, Crta., Madrid, Spain.
Chiloeches
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A
García
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N
Montes
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Toro
M J
MJ
D000818:Animals; D000924:Anticholesteremic Agents / Q000494:pharmacology*; D002460:Cell Line; D015500:Chloramphenicol O-Acetyltransferase / Q000235:genetics; D005576:Colforsin / Q000494:pharmacology; D000242:Cyclic AMP / Q000378:metabolism*; D019204:GTP-Binding Proteins / Q000378:metabolism; D015870:Gene Expression / Q000187:drug effects; D017930:Genes, Reporter / Q000187:drug effects; D013006:Growth Hormone / Q000235:genetics* / Q000378:metabolism; D007700:Kinetics; D008148:Lovastatin / Q000494:pharmacology*; D008798:Mevalonic Acid / Q000494:pharmacology; D010902:Pituitary Gland / Q000187:drug effects / Q000378:metabolism; D011388:Prolactin / Q000235:genetics* / Q000378:metabolism; D011401:Promoter Regions, Genetic / Q000187:drug effects; D051381:Rats; D015398:Signal Transduction
D000924:Anticholesteremic Agents; D005576:Colforsin; D011388:Prolactin; D013006:Growth Hormone; D008148:Lovastatin; D000242:Cyclic AMP; D015500:Chloramphenicol O-Acetyltransferase; D019204:GTP-Binding Proteins; D008798:Mevalonic Acid