Role of CXC chemokine ligand 13 in oral squamous cell carcinoma associated osteolysis in athymic mice

Oral squamous cell carcinomas (OSCC) are malignant tumors with a potent activity of local bone invasion; however, the molecular mechanisms of tumor osteolysis are unclear. In this study, we identified high level expression of chemokine ligand, CXCL13 and RANK ligand (RANKL) in OSCC cells (SCC1, SCC1...

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Veröffentlicht in:International journal of cancer 2010-05, Vol.126 (10), p.2319-2329
Hauptverfasser: Pandruvada, Subramanya N.M., Yuvaraj, Sambandam, Liu, Xiang, Sundaram, Kumaran, Shanmugarajan, Srinivasan, Ries, William L., Norris, James S., London, Steven D., Reddy, Sakamuri V.
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container_issue 10
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container_title International journal of cancer
container_volume 126
creator Pandruvada, Subramanya N.M.
Yuvaraj, Sambandam
Liu, Xiang
Sundaram, Kumaran
Shanmugarajan, Srinivasan
Ries, William L.
Norris, James S.
London, Steven D.
Reddy, Sakamuri V.
description Oral squamous cell carcinomas (OSCC) are malignant tumors with a potent activity of local bone invasion; however, the molecular mechanisms of tumor osteolysis are unclear. In this study, we identified high level expression of chemokine ligand, CXCL13 and RANK ligand (RANKL) in OSCC cells (SCC1, SCC12 and SCC14a). OSCC cell‐conditioned media (20%) induced osteoclast differentiation which was inhibited by OPG in peripheral blood monocyte cultures indicating that OSCC cells produce soluble RANKL. Recombinant hCXCL13 (10 ng/ml) significantly enhanced RANKL‐stimulated osteoclast differentiation in these cultures. Trans‐well migration assay identified that CXCL13 induces chemotaxis of peripheral blood monocytes in vitro which was inhibited by addition of anti‐CXCR5 receptor antibody. Zymogram analysis of conditioned media from OSCC cells revealed matrix metalloproteinase‐9 (MMP‐9) activity. Interestingly, CXCL13 treatment to OSCC cells induced CXCR5 and MMP‐9 expression suggesting an autocrine regulatory function in OSCC cells. To examine the OSCC tumor cell bone invasion/osteolysis, we established an in vivo model for OSCC by subcutaneous injection of OSCC cells onto the surface of calvaria in NCr‐nu/nu athymic mice, which developed tumors in 4–5 weeks. μCT analysis revealed numerous osteolytic lesions in calvaria from OSCC tumor‐bearing mice. Histochemical staining of calvarial sections from these mice revealed a significant increase in the numbers of TRAP‐positive osteoclasts at the tumor‐bone interface. Immunohistochemical analysis confirmed CXCL13 and MMP‐9 expression in tumor cells. Thus, our data implicate a functional role for CXCL13 in bone invasion and may be a potential therapeutic target to prevent osteolysis associated with OSCC tumors in vivo.
doi_str_mv 10.1002/ijc.24920
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In this study, we identified high level expression of chemokine ligand, CXCL13 and RANK ligand (RANKL) in OSCC cells (SCC1, SCC12 and SCC14a). OSCC cell‐conditioned media (20%) induced osteoclast differentiation which was inhibited by OPG in peripheral blood monocyte cultures indicating that OSCC cells produce soluble RANKL. Recombinant hCXCL13 (10 ng/ml) significantly enhanced RANKL‐stimulated osteoclast differentiation in these cultures. Trans‐well migration assay identified that CXCL13 induces chemotaxis of peripheral blood monocytes in vitro which was inhibited by addition of anti‐CXCR5 receptor antibody. Zymogram analysis of conditioned media from OSCC cells revealed matrix metalloproteinase‐9 (MMP‐9) activity. Interestingly, CXCL13 treatment to OSCC cells induced CXCR5 and MMP‐9 expression suggesting an autocrine regulatory function in OSCC cells. To examine the OSCC tumor cell bone invasion/osteolysis, we established an in vivo model for OSCC by subcutaneous injection of OSCC cells onto the surface of calvaria in NCr‐nu/nu athymic mice, which developed tumors in 4–5 weeks. μCT analysis revealed numerous osteolytic lesions in calvaria from OSCC tumor‐bearing mice. Histochemical staining of calvarial sections from these mice revealed a significant increase in the numbers of TRAP‐positive osteoclasts at the tumor‐bone interface. Immunohistochemical analysis confirmed CXCL13 and MMP‐9 expression in tumor cells. 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To examine the OSCC tumor cell bone invasion/osteolysis, we established an in vivo model for OSCC by subcutaneous injection of OSCC cells onto the surface of calvaria in NCr‐nu/nu athymic mice, which developed tumors in 4–5 weeks. μCT analysis revealed numerous osteolytic lesions in calvaria from OSCC tumor‐bearing mice. Histochemical staining of calvarial sections from these mice revealed a significant increase in the numbers of TRAP‐positive osteoclasts at the tumor‐bone interface. Immunohistochemical analysis confirmed CXCL13 and MMP‐9 expression in tumor cells. Thus, our data implicate a functional role for CXCL13 in bone invasion and may be a potential therapeutic target to prevent osteolysis associated with OSCC tumors in vivo.</abstract><cop>Hoboken</cop><pub>Wiley Subscription Services, Inc., A Wiley Company</pub><pmid>19816883</pmid><doi>10.1002/ijc.24920</doi><tpages>11</tpages><oa>free_for_read</oa></addata></record>
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subjects Animals
Biological and medical sciences
Blotting, Western
Carcinoma, Squamous Cell - enzymology
Carcinoma, Squamous Cell - metabolism
Cell Differentiation
Cell Line, Tumor
chemokine
Chemokine CXCL13 - genetics
Chemokine CXCL13 - metabolism
Chemotaxis
Disease Models, Animal
Gene Expression Regulation, Neoplastic
Humans
Immunohistochemistry
Matrix Metalloproteinase 9 - metabolism
Medical sciences
Mice
Mice, Nude
MMP‐9, bone invasion
mouse model
Mouth Neoplasms - enzymology
Mouth Neoplasms - metabolism
oral squamous cell carcinoma (OSCC)
Osteolysis - enzymology
Osteolysis - metabolism
Otorhinolaryngology. Stomatology
RANK Ligand - metabolism
RANK ligand, osteoclast
Reverse Transcriptase Polymerase Chain Reaction
Tumors
Upper respiratory tract, upper alimentary tract, paranasal sinuses, salivary glands: diseases, semeiology
title Role of CXC chemokine ligand 13 in oral squamous cell carcinoma associated osteolysis in athymic mice
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