Intracellular calcium and protein kinase C mediate expression of receptor activator of nuclear factor-κB ligand and osteoprotegerin in osteoblasts

被引:78
作者
Takami, M
Takahashi, N
Udagawa, N
Miyaura, C
Suda, K
Woo, JT
Martin, TJ
Nagai, K
Suda, T
机构
[1] Showa Univ, Sch Dent, Dept Biochem, Shinagawa Ku, Tokyo 1428555, Japan
[2] Tokyo Inst Technol, Dept Bioengn, Yokohama, Kanagawa 2268501, Japan
[3] Tokyo Univ Pharm & Life Sci, Sch Pharm, Dept Biochem, Tokyo 1920392, Japan
[4] St Vincents Inst Med Res, Fitzroy, Vic 3065, Australia
关键词
D O I
10.1210/en.141.12.4711
中图分类号
R5 [内科学];
学科分类号
1002 ; 100201 ;
摘要
Receptor activator of nuclear factor-kappaB ligand (RANKL) and osteoprotegerin (OPG) produced by osteoblasts/stromal cells are involved as positive and negative regulators in osteoclast formation. Three independent signals have been proposed to induce RANKL expression in osteoblasts/stromal cells: vitamin D receptor-, cAMP-, and gp130-mediated signals. We previously reported that intracellular calcium-elevating compounds such as ionomycin, cyclopiazonic acid, and thapsigargin induced osteoclast formation in cocultures of mouse bone marrow cells and primary osteoblasts. Increases in calcium concentration in culture medium also induced osteoclast formation in cocultures. Treatment of primary osteoblasts with these compounds or with high calcium medium stimulated the expression of both RANKL and OPG messenger RNAs (mRNAs). 1,2-Bis(o-aminophenoxy)ethane-N,N,N',N'-tetraacetic acid)-tetra(acetoxymethyl)ester, an intracellular calcium chelator, suppressed both ionamycin-induced osteoclast formation in cocultures and expression of RANKL and OPG mRNAs in primary osteoblasts. Phorbol 12-myristate 13-acetate (PMA), an activator of protein kinase C, also stimulated osteoclast formation in these cocultures and the expression of RANKL and OPG mRNAs in primary osteoblasts. Protein kinase C inhibitors such as calphostin and staurosporin suppressed ionomycin- and PMA-induced osteoclast formation in cocultures and expression of RANKL and OPG mRNAs in primary osteoblasts. Ionomycin stimulated RANKL mRNA expression in ST2 and MC3T3-G2/PA6 cells, but not in MC3T3-E1 or NIH-3T3 cells. These effects were closely correlated with osteoclast formation in response to ionomycin in cocultures with these stromal cell lines. OPG strongly inhibited osteoclast formation induced by calcium-elevating compounds and PMA in cocultures, suggesting that RANKL expression in osteoblasts is a rate-limiting step for osteoclast induction. Forskolin, an activator of cAMP signals, also stimulated osteoclast formation in cocultures. Forskolin enhanced RANKL mRNA expression but suppressed OPG mRNA expression in primary osteoblasts. These results suggest that the calcium/protein kinase C signal in osteoblasts/stromal cells is the fourth signal for inducing RANKL mRNA expression, which, in turn, stimulates osteoclast formation.
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页码:4711 / 4719
页数:9
相关论文
共 38 条
[31]   Stimulation of osteoclast formation by 1,25-dihydroxyvitamin D requires its binding to vitamin D receptor (VDR) in osteoblastic cells: Studies using VDR knockout mice [J].
Takeda, S ;
Yoshizawa, T ;
Nagai, Y ;
Yamato, H ;
Fukumoto, S ;
Sekine, K ;
Kato, S ;
Matsumoto, T ;
Fujita, T .
ENDOCRINOLOGY, 1999, 140 (02) :1005-1008
[32]   THE BONE MARROW-DERIVED STROMAL CELL-LINES MC3T3-G2/PA6 AND ST2 SUPPORT OSTEOCLAST-LIKE CELL-DIFFERENTIATION IN COCULTURES WITH MOUSE SPLEEN-CELLS [J].
UDAGAWA, N ;
TAKAHASHI, N ;
AKATSU, T ;
SASAKI, T ;
YAMAGUCHI, A ;
KODAMA, H ;
MARTIN, TJ ;
SUDA, T .
ENDOCRINOLOGY, 1989, 125 (04) :1805-1813
[33]   INTERLEUKIN (IL)-6 INDUCTION OF OSTEOCLAST DIFFERENTIATION DEPENDS ON IL-6 RECEPTORS EXPRESSED ON OSTEOBLASTIC CELLS BUT NOT ON OSTEOCLAST PROGENITORS [J].
UDAGAWA, N ;
TAKAHASHI, N ;
KATAGIRI, T ;
TAMURA, T ;
WADA, S ;
FINDLAY, DM ;
MARTIN, TJ ;
HIROTA, H ;
TADA, T ;
KISHIMOTO, T ;
SUDA, T .
JOURNAL OF EXPERIMENTAL MEDICINE, 1995, 182 (05) :1461-1468
[34]   BONE - FORMATION BY AUTOINDUCTION [J].
URIST, MR .
SCIENCE, 1965, 150 (3698) :893-&
[35]   CONTROL OF BONE-RESORPTION BY HEMATOPOIETIC TISSUE - INDUCTION AND REVERSAL OF CONGENITAL OSTEOPETROSIS IN MICE THROUGH USE OF BONE-MARROW AND SPLENIC TRANSPLANTS [J].
WALKER, DG .
JOURNAL OF EXPERIMENTAL MEDICINE, 1975, 142 (03) :651-663
[36]   TRANCE is a novel ligand of the tumor necrosis factor receptor family that activates c-Jun N-terminal kinase in T cells [J].
Wong, BR ;
Rho, JR ;
Arron, J ;
Robinson, E ;
Orlinick, J ;
Chao, M ;
Kalachikov, S ;
Cayani, E ;
Bartlett, FS ;
Frankel, WN ;
Lee, SY ;
Choi, YW .
JOURNAL OF BIOLOGICAL CHEMISTRY, 1997, 272 (40) :25190-25194
[37]   Identity of osteoclastogenesis inhibitory factor (OCIF) and osteoprotegerin (OPG):: A mechanism by which OPG/OCIF inhibits osteoclastogenesis in vitro [J].
Yasuda, H ;
Shima, N ;
Nakagawa, N ;
Mochizuki, SI ;
Yano, K ;
Fujise, N ;
Sato, Y ;
Goto, M ;
Yamaguchi, K ;
Kuriyama, M ;
Kanno, T ;
Murakami, A ;
Tsuda, E ;
Morinaga, T ;
Higashio, K .
ENDOCRINOLOGY, 1998, 139 (03) :1329-1337
[38]   Osteoclast differentiation factor is a ligand for osteoprotegerin osteoclastogenesis-inhibitory factor and is identical to TRANCE/RANKL [J].
Yasuda, H ;
Shima, N ;
Nakagawa, N ;
Yamaguchi, K ;
Kinosaki, M ;
Mochizuki, S ;
Tomoyasu, A ;
Yano, K ;
Goto, M ;
Murakami, A ;
Tsuda, E ;
Morinaga, T ;
Higashio, K ;
Udagawa, N ;
Takahashi, N ;
Suda, T .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1998, 95 (07) :3597-3602