Changes in cytoskeletal proteins in response to parathyroid hormone and 1,25-dihydroxyvitamin D in human osteoblastic...

We have examined the influence of parathyroid hormone (PTH) and 1,25(OH)2 vitamin D (1,25(OH)2D) on cytoskeletal assembly and biosynthesis in relation with cAMP production and parameters of cell growth and differentiation in normal human osteoblastic cells. Untreated human bone cells showed elongated morphology associated with high levels of actin, vimentin, alpha- and beta-tubulins and alpha-actinin as determined by 2-dimensional-gel electrophoresis and [35S]methionine labelling of cytoskeletal proteins. PTH (20 nM, 24 h) decreased the de novo biosynthesis of vimentin and alpha-actinin in human bone cells, an effect associated with a rise in intracellular cyclic AMP. In addition, PTH induced cytoskeletal disassembly as shown by a 52-70% decrease in the Triton-insoluble fractions of actin, alpha-tubulins and alpha-actinin. 1,25(OH)2D (10 nM, 24 h) also induced a 40-64% decrease in the polymerized fractions of actin, alpha-tubulins and alpha-actinin. These changes were associated with an 83% increase in osteocalcin production. Under these conditions, neither PTH nor 1,25(OH)2D at the doses tested affected alkaline phosphatase activity or cell growth as assessed by [3H]thymidine incorporation into DNA. The results show that PTH and 1,25(OH)2D induce similar inhibition of cytoskeletal proteins assembly involving microfilaments and microtubules in human osteoblastic cells. These alterations of cytoskeletal arrangement in response to PTH and 1,25(OH)2D may contribute to the functional response of human osteoblastic cells to these bone-resorbing hormones.