Evidence for direct effects of prolactin on human osteoblasts: Inhibition of cell growth and mineralization.
Hyperprolactinemia is one of the risk factor of decrease in bone mass which has been believed to be mediated by hypogonadism. However, the presence of prolactin receptor in human osteosarcoma cell line and primary bone cell culture from mouse calvariae supported the hypothesis of a direct prolactin (PRL) action on bone cells. Therefore, the aim of this study was to investigate the role of PRL and its signal transduction pathway in the regulation of bone metabolism via osteoblast differentiation. Human pre-osteoblasts (SV-HFO) that differentiate in a 3-week period from proliferating pre-osteoblasts (days 2-7) to extracellular matrix producing cells (days 7-14) which is eventually mineralized (days 14-21) were used. Concentration of PRL mimicked a lactating period (100 ng/ml) was used to incubate SV-HFO for 21 days in osteogenic medium. Human prolactin receptor mRNA and protein are expressed in SV-HFO. PRL significantly decreased osteoblast number (DNA content) which was due to a decrease in proliferation. PRL increased osteogenic markers, RUNX2 and ALP in early stage of osteoblast differentiation while decreasing it later suggesting a bi-directional effect. Calcium measurement and Alizarin red staining showed a reduction of mineralization by PRL while having neither an effect on osteoblast activity nor RANKL/OPG mRNA ratio. We also demonstrated that PRL action on mineralization was not via PI-3 kinase pathway. The present study provides evidence of a direct effect of prolactin on osteoblast differentiation and in vitro mineralization.
Seriwatanachai
Dutmanee
D
Department of Physiology, Mahidol University, Bangkok, Thailand.
Krishnamra
Nateetip
N
van Leeuwen
J P T M
JP
D015415:Biomarkers / Q000032:analysis; D001842:Bone and Bones / Q000166:cytology / Q000378:metabolism; D002113:Calcification, Physiologic / Q000187:drug effects*; D002452:Cell Count; D002454:Cell Differentiation; D049109:Cell Proliferation / Q000187:drug effects*; D002478:Cells, Cultured; D006801:Humans; D010006:Osteoblasts / Q000166:cytology* / Q000187:drug effects; D011388:Prolactin / Q000494:pharmacology*; D015398:Signal Transduction
D015415:Biomarkers; D011388:Prolactin