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Am J Physiol Endocrinol Metab (November 12, 2002). doi:10.1152/ajpendo.00307.2002
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Articles in PresS, published online ahead of print November 12, 2002
Am J Physiol Endocrinol Metab, 10.1152/ajpendo.00307.2002
Submitted on July 11, 2002
Accepted on October 31, 2002

Human osteoblast-like cell proliferation induced by calcitonin related peptides involves PKC activity

Isabella Villa1, Chiara Dal Fiume1, Anna Maestroni2, Alessandro Rubinacci1, Flavio Ravasi3, and Francesca Guidobono4*

1 Bone Metabolic Unit, Scientific Institute H. San Raffaele, Milan, Italy
2 Laboratory of Renal Pathophysiology, Division of Medicine, Scientific Institute H. San Raffaele, Milan, Italy
3 Department of Orthopaedics, Scientific Institute H. San Raffaele, Milan, Italy
4 Department of Pharmacology, Chemotherapy and Medical Toxicology, University of Milan, Milan, Italy

* To whom correspondence should be addressed. E-mail: Francesca.Guidobono{at}unimi.it.

The calcitonin peptides (CT, CGRP, amylin) share many biological actions including activity on bone cells. In the present study, CT (10-11-10-9M) stimulated [3H]-thymidine incorporation in primary culture of human osteoblasts (hOB) as already demonstrated for CGRP and amylin. RT-PCR analysis showed that the calcitonin receptor and the calcitonin receptor-like receptor are both expressed in hOB. In these cells CT (10-10M) and amylin (10-9M), in contrast with CGRP (10-8M), did not increase cAMP production. All three peptides stimulated PKC activity. In order to evaluate PKC involvement in hOB proliferation, cells were incubated with phorbol12,13-dibutyrate, a stimulator of PKC activity; cell proliferation was increased in a dose dependent manner (EC50 = 3.4 x 10-8M). Staurosporine (10-9M), a PKC inhibitor, blocked phorbol12,13-dibutyrate induced PKC activity and cell proliferation. Inhibition of PKC by staurosporine also counteracted the stimulatory effect of CT, CGRP and amylin on hOB proliferation. From these data it is deduced that the activation of PKC is important for hOB proliferation and that it is involved in the anabolic effect of CT peptides on bone.




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