PMID- 28786489 OWN - NLM STAT- MEDLINE DCOM- 20171212 LR - 20181202 IS - 1097-4652 (Electronic) IS - 0021-9541 (Linking) VI - 233 IP - 3 DP - 2018 Mar TI - Spleen tyrosine kinase influences the early stages of multilineage differentiation of bone marrow stromal cell lines by regulating phospholipase C gamma activities. PG - 2549-2559 LID - 10.1002/jcp.26130 [doi] AB - Bone marrow stromal cells (BMSCs) are multipotent cells that can differentiate into adipocytes and osteoblasts. Inadequate BMSC differentiation is occasionally implicated in chronic bone metabolic disorders. However, specific signaling pathways directing BMSC differentiation have not been elucidated. Here, we explored the roles of spleen tyrosine kinase (Syk) in BMSC differentiation into adipocytes and osteoblasts. We found that Syk phosphorylation was increased in the early stage, whereas its protein expression was gradually decreased during the adipogenic and osteogenic differentiation of two mouse mesenchymal stromal cell lines, ST2 and 10T(1/2), and a human BMSC line, UE6E-7-16. Syk inactivation with either a pharmacological inhibitor or Syk-specific siRNA suppressed adipogenic differentiation, characterized by decreased lipid droplet appearance and the gene expression of fatty acid protein 4 (Fabp4), peroxisome proliferator-activated receptor gamma2 (Pparg2), CCAAT/enhancer binding proteins alpha (C/EBPalpha), and C/EBPbeta. In contrast, Syk inhibition promoted osteogenic differentiation, represented by increase in matrix mineralization and alkaline phosphatase (ALP) activity, as well as the expression levels of osteocalcin, runt-related transcription factor 2 (Runx2), and distal-less homeobox 5 (Dlx5) mRNAs. We also found that Syk-induced signals are mediated by phospholipase C gamma1 (PLCgamma1) in osteogenesis and PLCgamma2 in adipogenesis. Notably, Syk-activated PLCgamma2 signaling was partly modulated through B-cell linker protein (BLNK) in adipogenic differentiation. On the other hand, growth factor receptor-binding protein 2 (Grb2) was involved in Syk-PLCgamma1 axis in osteogenic differentiation. Taken together, these results indicate that Syk-PLCgamma signaling has a dual role in regulating the initial stage of adipogenic and osteogenic differentiation of BMSCs. CI - (c) 2017 Wiley Periodicals, Inc. FAU - Kusuyama, Joji AU - Kusuyama J AUID- ORCID: 0000-0001-9588-8089 AD - Department of Oral Biochemistry, Field of Developmental Medicine, Kagoshima University Graduate School of Medical and Dental Sciences, Kagoshima, Japan. FAU - Kamisono, Ai AU - Kamisono A AD - Department of Oral Biochemistry, Field of Developmental Medicine, Kagoshima University Graduate School of Medical and Dental Sciences, Kagoshima, Japan. FAU - ChangHwan, Seong AU - ChangHwan S AD - Department of Oral Biochemistry, Field of Developmental Medicine, Kagoshima University Graduate School of Medical and Dental Sciences, Kagoshima, Japan. FAU - Amir, Muhammad S AU - Amir MS AD - Department of Oral Biochemistry, Field of Developmental Medicine, Kagoshima University Graduate School of Medical and Dental Sciences, Kagoshima, Japan. AD - Department of Oral and Maxillofacial Surgery, Field of Oral and Maxillofacial Rehabilitation, Kagoshima University Graduate School of Medical and Dental Sciences, Kagoshima, Japan. AD - Department of Oral and Maxillofacial Surgery, Faculty of Dentistry, Airlangga University, Surabaya, Indonesia. AD - Campus A UNAIR, JL, Mayjen Professor Doktor Moestopo, Pacar Kembang, Tambaksari, Kota SBY,, Jawa Timur, Indonesia. FAU - Bandow, Kenjiro AU - Bandow K AD - Department of Oral Biology and Tissue Engineering, Meikai University School of Dentistry, Saitama, Japan. FAU - Eiraku, Nahoko AU - Eiraku N AD - Department of Periodontology, Field of Oral and Maxillofacial Rehabilitation, Kagoshima University Graduate School of Medical and Dental Sciences, Kagoshima, Japan. FAU - Ohnishi, Tomokazu AU - Ohnishi T AD - Department of Oral Biochemistry, Field of Developmental Medicine, Kagoshima University Graduate School of Medical and Dental Sciences, Kagoshima, Japan. FAU - Matsuguchi, Tetsuya AU - Matsuguchi T AD - Department of Oral Biochemistry, Field of Developmental Medicine, Kagoshima University Graduate School of Medical and Dental Sciences, Kagoshima, Japan. LA - eng PT - Journal Article DEP - 20170928 PL - United States TA - J Cell Physiol JT - Journal of cellular physiology JID - 0050222 RN - 0 (Adaptor Proteins, Signal Transducing) RN - 0 (B cell linker protein) RN - 0 (Biomarkers) RN - 0 (GRB2 Adaptor Protein) RN - 0 (GRB2 protein, human) RN - 0 (Grb2 protein, mouse) RN - 0 (RNA, Messenger) RN - EC 2.7.10.2 (SYK protein, human) RN - EC 2.7.10.2 (Syk Kinase) RN - EC 2.7.10.2 (Syk protein, mouse) RN - EC 3.1.4.11 (PLCG1 protein, human) RN - EC 3.1.4.3 (Phospholipase C gamma) RN - EC 3.1.4.3 (Plcg1 protein, mouse) SB - IM MH - Adaptor Proteins, Signal Transducing/genetics/metabolism MH - Adipocytes/*enzymology MH - *Adipogenesis MH - Animals MH - Biomarkers/metabolism MH - Cell Line MH - *Cell Lineage MH - GRB2 Adaptor Protein/genetics/metabolism MH - Gene Expression Regulation MH - Humans MH - Mesenchymal Stem Cells/*enzymology MH - Mice, Inbred C3H MH - Osteoblasts/*enzymology MH - *Osteogenesis MH - Phenotype MH - Phospholipase C gamma/genetics/*metabolism MH - Phosphorylation MH - RNA Interference MH - RNA, Messenger/genetics/metabolism MH - Signal Transduction MH - Syk Kinase/genetics/*metabolism MH - Time Factors MH - Transfection OTO - NOTNLM OT - PLC OT - Syk OT - adipogenesis OT - bone marrow stromal cell OT - osteogenesis EDAT- 2017/08/09 06:00 MHDA- 2017/12/13 06:00 CRDT- 2017/08/09 06:00 PHST- 2017/05/14 00:00 [received] PHST- 2017/08/01 00:00 [accepted] PHST- 2017/08/09 06:00 [pubmed] PHST- 2017/12/13 06:00 [medline] PHST- 2017/08/09 06:00 [entrez] AID - 10.1002/jcp.26130 [doi] PST - ppublish SO - J Cell Physiol. 2018 Mar;233(3):2549-2559. doi: 10.1002/jcp.26130. Epub 2017 Sep 28.