title: Osteoclast differentiation gene expression profiling reveals chemokine CCL4 mediates RANKL-induced osteoclast migration and invasion via PI3K pathway.
abstract: The migration of osteoclasts (OCs) from circulation and bone marrow into bone surface plays a critical role in the pathogenesis of some bone resorptive diseases, such as rheumatoid arthritis and osteoporosis. To date, how the migration of OCs remains unclear. We investigated gene expression profiling in osteoclastic differentiation of bone marrow-derived macrophages (BMMs) into OCs by microarray analysis. We identified 387 genes overexpressed in osteoclastic differentiation of BMMs. Among them, chemokine CCL4 showed a robust up-regulation signal. High expression of CCL4 was validated in primary BMMs and OC precursor cell line RAW264.7 during differentiation into OCs. The CCL4 neutralization decreased RANKL-induced OC precursor cell migration and invasion in Matrigel-coated transwell membranes assay and in vitro wound healing assay. However, CCL4 inhibition did not affect OCs differentiation and differentiation associated gene expression. The CCL4 inhibition promoted the PI3K phosphorylation at 45 to 60 minutes after RANKL stimulation in RAW264.7. This study indicated that chemokine CCL4 is an important regulator for OCs migration via PI3K pathway, providing a novel therapy target for bone resorptive diseases.
authors.affiliation: Department of Rheumatology, The First Affiliated Hospital of Nanjing Medical University, Nanjing, China.
authors.forename: Wenhua
authors.initials: W
authors.lastname: Xuan
authors.affiliation: Department of Traditional Chinese Medicine, The First Affiliated Hospital of Nanjing Medical University, Nanjing, China.
authors.forename: Xiaoke
authors.initials: X
authors.lastname: Feng
authors.affiliation: Department of Rheumatology, The First Affiliated Hospital of Nanjing Medical University, Nanjing, China.
authors.forename: Chen
authors.initials: C
authors.lastname: Qian
authors.affiliation: Department of Rheumatology, The First Affiliated Hospital of Nanjing Medical University, Nanjing, China.
authors.forename: Liuying
authors.initials: L
authors.lastname: Peng
authors.affiliation: Department of Rheumatology, The First Affiliated Hospital of Nanjing Medical University, Nanjing, China.
authors.forename: Yumeng
authors.initials: Y
authors.lastname: Shi
authors.affiliation: Department of Rheumatology, The First Affiliated Hospital of Nanjing Medical University, Nanjing, China.
authors.forename: Lingxiao
authors.initials: L
authors.lastname: Xu
authors.affiliation: Department of Cardiology, The First Affiliated Hospital of Nanjing Medical University, Nanjing, China.
authors.forename: Fang
authors.initials: F
authors.lastname: Wang
authors.affiliation: Department of Rheumatology, The First Affiliated Hospital of Nanjing Medical University, Nanjing, China.
authors.forename: Wenfeng
authors.identifier: http://orcid.org/0000-0002-5160-2023
authors.initials: W
authors.lastname: Tan
mesh_terms: D000818:Animals; D017209:Apoptosis; D015153:Blotting, Western; D002448:Cell Adhesion; D002454:Cell Differentiation / Q000235:genetics*; D002465:Cell Movement; D049109:Cell Proliferation; D002478:Cells, Cultured; D054407:Chemokine CCL4 / Q000235:genetics* / Q000378:metabolism; D020869:Gene Expression Profiling; D008264:Macrophages / Q000378:metabolism / Q000473:pathology; D051379:Mice; D008813:Mice, Inbred ICR; D046228:Microarray Analysis; D010010:Osteoclasts / Q000378:metabolism / Q000473:pathology*; D019869:Phosphatidylinositol 3-Kinases / Q000235:genetics / Q000378:metabolism*; D053245:RANK Ligand / Q000235:genetics* / Q000378:metabolism; D012333:RNA, Messenger / Q000235:genetics; D060888:Real-Time Polymerase Chain Reaction; D020133:Reverse Transcriptase Polymerase Chain Reaction; D015398:Signal Transduction
chemical_list: D054407:Chemokine CCL4; D053245:RANK Ligand; D012333:RNA, Messenger; C505689:Tnfsf11 protein, mouse; D019869:Phosphatidylinositol 3-Kinases
keywords: CCL4; chemokine; migration; osteoclast