caa a22 4500 605463638 CHVBK 20210128100254.0 cr unu---uuuuu 210128e20150301xx s 000 0 eng 10.1007/s00774-014-0576-8 doi (NATIONALLICENCE)springer-10.1007/s00774-014-0576-8 Effects of interleukin-11 on the expression of human bone sialoprotein gene [Elektronische Daten] [Hiroyoshi Matsumura, Yohei Nakayama, Hideki Takai, Yorimasa Ogata] Interleukin-11 (IL-11) is a bone marrow stromal fibroblast-derived cytokine with a wide spectrum of activities in different biological systems. IL-11 and IL-6 are two cytokines known to rely on osteoblast-osteoclast communication for their effects on osteoclast differentiation. Bone sialoprotein (BSP) is a mineralized connective tissue-specific protein expressed in differentiated osteoblasts, odontoblasts, and cementoblasts. To determine the molecular basis of the transcriptional regulation of the human BSP gene by IL-11, we conducted real-time polymerase chain reactions (PCR), transient transfection analyses with chimeric constructs of the human BSP gene promoter linked to a luciferase reporter gene, gel mobility shift assays, and a chromatin immunoprecipitation assay using human osteoblast-like Saos2 cells. IL-11 (20ng/ml) increased BSP, Runx2, and Osterix mRNA levels at 6h and the alkaline phosphatase (ALP) mRNA level at 12h in osteoblast-like Saos2 cells. In a transient transfection assay, IL-11 (20ng/ml, 12h) increased luciferase activities of constructs between −60LUC and −868LUC including the human BSP gene promoter. Transcriptional stimulations by IL-11 were partially inhibited in the constructs that included 2-bp mutations in the cAMP response element 1 (CRE1, −72 to −79) and CRE2 (−667 to −674). When mutations were made in pairs of CRE1 and CRE2 in −868LUC, the effect of IL-11 on luciferase activity was almost totally abrogated. Transcriptional activities induced by IL-11 were inhibited by protein kinase A, tyrosine kinase, ERK1/2, and PI3-kinase inhibitors. Gel mobility shift analyses showed that IL-11 increased nuclear proteins binding to CRE1 and CRE2. CREB1, phospho-CREB1, c-Fos, and c-Jun antibodies disrupted the formation of CRE1 and CRE2 protein complexes. These data demonstrate that IL-11 stimulates BSP gene transcription via CRE1 and CRE2 elements in the human BSP gene promoter. The Japanese Society for Bone and Mineral Research and Springer Japan, 2014 Bone sialoprotein nationallicence Interleukin-11 nationallicence Osteoblasts nationallicence Transcription nationallicence Matsumura Hiroyoshi Department of Periodontology, Nihon University School of Dentistry at Matsudo, 271-8587, Matsudo, Chiba, Japan aut Nakayama Yohei Department of Periodontology and Research Institute of Oral Science, Nihon University School of Dentistry at Matsudo, 271-8587, Matsudo, Chiba, Japan aut Takai Hideki Department of Periodontology and Research Institute of Oral Science, Nihon University School of Dentistry at Matsudo, 271-8587, Matsudo, Chiba, Japan aut Ogata Yorimasa Department of Periodontology and Research Institute of Oral Science, Nihon University School of Dentistry at Matsudo, 271-8587, Matsudo, Chiba, Japan aut Journal of Bone and Mineral Metabolism Springer Japan 33/2(2015-03-01), 142-153 0914-8779 33:2<142 2015 33 774 https://doi.org/10.1007/s00774-014-0576-8 text/html Onlinezugriff via DOI BK010053 XK010053 XK010000 Metadata rights reserved Springer special CC-BY-NC licence nationallicence 1 research-article jats NATIONALLICENCE NATIONALLICENCE NL-springer NATIONALLICENCE 856 40 https://doi.org/10.1007/s00774-014-0576-8 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Matsumura Hiroyoshi Department of Periodontology, Nihon University School of Dentistry at Matsudo, 271-8587, Matsudo, Chiba, Japan aut NATIONALLICENCE 700 1- Nakayama Yohei Department of Periodontology and Research Institute of Oral Science, Nihon University School of Dentistry at Matsudo, 271-8587, Matsudo, Chiba, Japan aut NATIONALLICENCE 700 1- Takai Hideki Department of Periodontology and Research Institute of Oral Science, Nihon University School of Dentistry at Matsudo, 271-8587, Matsudo, Chiba, Japan aut NATIONALLICENCE 700 1- Ogata Yorimasa Department of Periodontology and Research Institute of Oral Science, Nihon University School of Dentistry at Matsudo, 271-8587, Matsudo, Chiba, Japan aut NATIONALLICENCE 773 0- Journal of Bone and Mineral Metabolism Springer Japan 33/2(2015-03-01), 142-153 0914-8779 33:2<142 2015 33 774