Mutant monocyte chemoattractant protein-1 protein (7ND) inhibits osteoclast differentiation and reduces oral squamous carcinoma cell bone invasion

Luo,Shuyu; Zhou,Chuanxiang; Zhang,Jianming; Chen,Mengshan; Li,Hongjie; Zheng,Shanchuan; Quan,Jingjing

doi:10.3892/ol.2018.8308

Mutant monocyte chemoattractant protein-1 protein (7ND) inhibits osteoclast differentiation and reduces oral squamous carcinoma cell bone invasion

Authors:
- Shuyu Luo
- Chuanxiang Zhou
- Jianming Zhang
- Mengshan Chen
- Hongjie Li
- Shanchuan Zheng
- Jingjing Quan
View Affiliations

Affiliations: Department of Oral Pathology, School and Hospital of Stomatology, Tianjin Medical University, Heping, Tianjin 300070, P.R. China, Department of Oral Pathology, Peking University School and Hospital of Stomatology, Haidian, Beijing 100081, P.R. China, Department of Stomatology, General Hospital of Tianjin Medical University, Heping, Tianjin 300052, P.R. China, School of Public Health, Tianjin Medical University, Heping, Tianjin 300070, P.R. China, Department of Stomatology, WuQing People's Hospital, Wuqing, Tianjin 301700, P.R. China, Guanghua School of Stomatology, Hospital of Stomatology, Sun Yat‑Sen University and Guangdong Provincial Key Laboratory of Stomatology, Guangzhou, Guangdong 510080, P.R. China
Published online on: March 21, 2018 https://doi.org/10.3892/ol.2018.8308
Pages: 7760-7768
Copyright: © Luo et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

Metrics: Total Views: 0 (Spandidos Publications: | PMC Statistics: )

Metrics: Total PDF Downloads: 0 (Spandidos Publications: | PMC Statistics: )

Cited By (CrossRef): 0 citations Loading Articles...

Abstract

The seven‑amino acid truncated (7ND) protein is an N‑terminal deletion mutant of monocyte chemoattractant protein‑1 (MCP‑1) and it functions as a dominant‑negative inhibitor. 7ND and wild‑type MCP‑1 form a heterodimer, which binds to MCP‑1 receptors and inhibits monocyte chemotaxis. In the present study, the 7ND protein was cloned, expressed and purified. An MTT assay revealed that the proliferation of oral squamous cell carcinoma (OSCC) SCC25 cells was not affected following 3 days of treatment with synthetic 7ND protein. Serial dilutions of the 7ND protein were tested for monocyte migration and osteoclast differentiation, and tartrate‑resistant acid phosphatase staining demonstrated that significantly fewer osteoclasts were differentiated from cluster of differentiation 14+ (CD14+) monocytes using magnetic activated cell sorting. Immunofluorescence confirmed these results and significantly less F‑actin staining was observed in 7ND‑treated osteoclasts. Furthermore, bone invasion was examined by subcutaneously injecting SCC25 cells into the area overlaying the calvariae of nude mice. The results demonstrated that the average tumor volume of SCC25 cells with 7ND protein was similar to the average volume of tumors formed by untreated SCC25 cells. Flow cytometric analysis suggested that the CD14+ subpopulation in the bone marrow of 7ND‑treated mice was reduced compared with that of untreated mice. Micro‑computed tomography imaging revealed significantly less bone resorption in the calvariae injected with SCC25 cells plus the 7ND protein. Taken together, the results of the present study demonstrated the potential therapeutic value of the 7ND protein. The 7ND MCP‑1 variant not only functions in vitro to inhibit osteoclast differentiation, but also reduces the progression of bone invasion by OSCC cells in vivo.

View Figures

View References

1	Lehmann MH, Torres-Domínguez LE, Price PJ, Brandmüller C, Kirschning CJ and Sutter G: CCL2 expression is mediated by type I IFN receptor and recruits NK and T cells to the lung during MVA infection. J Leukoc Biol. 99:1057–1064. 2016. View Article : Google Scholar : PubMed/NCBI
2	Qidwai T: Chemokine genetic polymorphism in human health and disease. Immunol Lett. 176:128–138. 2016. View Article : Google Scholar : PubMed/NCBI
3	Kim MS, Day CJ and Morrison NA: MCP-1 is induced by receptor activator of nuclear factor-{kappa}B ligand, promotes human osteoclast fusion, and rescues granulocyte macrophage colony-stimulating factor suppression of osteoclast formation. J Biol Chem. 280:16163–16169. 2005. View Article : Google Scholar : PubMed/NCBI
4	Kim MS, Day CJ, Selinger CI, Magno CL, Stephens SR and Morrison NA: MCP-1-induced human osteoclast-like cells are tartrate-resistant acid phosphatase, NFATc1, and calcitonin receptor-positive but require receptor activator of NFkappaB ligand for bone resorption. J Biol Chem. 281:1274–1285. 2006. View Article : Google Scholar : PubMed/NCBI
5	Morrison NA, Day CJ and Nicholson GC: Dominant negative MCP-1 blocks human osteoclast differentiation. J Cell Biochem. 115:303–312. 2014. View Article : Google Scholar : PubMed/NCBI
6	Quan J, Johnson NW, Zhou G, Parsons PG, Boyle GM and Gao J: Potential molecular targets for inhibiting bone invasion by oral squamous cell carcinoma: A review of mechanisms. Cancer Metastasis Rev. 31:209–219. 2012. View Article : Google Scholar : PubMed/NCBI
7	Quan J, Elhousiny M, Johnson NW and Gao J: Transforming growth factor-β1 treatment of oral cancer induces epithelial-mesenchymal transition and promotes bone invasion via enhanced activity of osteoclasts. Clin Exp Metastasis. 30:659–670. 2013. View Article : Google Scholar : PubMed/NCBI
8	Quan J, Morrison NA, Johnson NW and Gao J: MCP-1 as a potential target to inhibit the bone invasion by oral squamous cell carcinoma. J Cell Biochem. 115:1787–1798. 2014. View Article : Google Scholar : PubMed/NCBI
9	Gu J, Feng Y, Feng X, Sun C, Lei L, Ding W, Niu F, Jiao L, Yang M, Li Y, et al: Structural and biochemical characterization reveals LysGH15 as an unprecedented ‘EF-hand-like’ calcium-binding phage lysine. PLoS Pathog. 10:e10041092014. View Article : Google Scholar : PubMed/NCBI
10	Gong QM, Quan JJ, Jiang HW and Ling JQ: Regulation of the stromal cell-derived factor-1alpha-CXCR4 axis in human dental pulp cells. J Endod. 36:1499–1503. 2010. View Article : Google Scholar : PubMed/NCBI
11	Zhang Y, Ernst CA and Rollins BJ: MCP-1: Structure/activity analysis. Methods. 10:93–103. 1996. View Article : Google Scholar : PubMed/NCBI
12	Zhang Y and Rollins BJ: A dominant negative inhibitor indicates that monocyte chemoattractant protein 1 functions as a dimer. Mol Cell Biol. 15:4851–4855. 1995. View Article : Google Scholar : PubMed/NCBI
13	Ikeda Y, Yonemitsu Y, Kataoka C, Kitamoto S, Yamaoka T, Nishida K, Takeshita A, Egashira K and Sueishi K: Anti-monocyte chemoattractant protein-1 gene therapy attenuates pulmonary hypertension in rats. Am J Physiol Heart Circ Physiol. 283:H2021–H2028. 2002. View Article : Google Scholar : PubMed/NCBI
14	Ni W, Egashira K, Kitamoto S, Kataoka C, Koyanagi M, Inoue S, Imaizumi K, Akiyama C, Nishida KI and Takeshita A: New anti-monocyte chemoattractant protein-1 gene therapy attenuates atherosclerosis in apolipoprotein E-knockout mice. Circulation. 103:2096–2101. 2001. View Article : Google Scholar : PubMed/NCBI
15	Koga M, Kai H, Egami K, Murohara T, Ikeda A, Yasuoka S, Egashira K, Matsuishi T, Kai M, Kataoka Y, et al: Mutant MCP-1 therapy inhibits tumor angiogenesis and growth of malignant melanoma in mice. Biochem Biophys Res Commun. 365:279–284. 2008. View Article : Google Scholar : PubMed/NCBI
16	Yao Z, Keeney M, Lin TH, Pajarinen J, Barcay K, Waters H, Egashira K, Yang F and Goodman S: Mutant monocyte chemoattractant protein 1 protein attenuates migration of and inflammatory cytokine release by macrophages exposed to orthopedic implant wear particles. J Biomed Mater Res A. 102:3291–3297. 2014. View Article : Google Scholar : PubMed/NCBI
17	Jiang X, Sato T, Yao Z, Keeney M, Pajarinen J, Lin TH, Loi F, Egashira K, Goodman S and Yang F: Local delivery of mutant CCL2 protein-reduced orthopaedic implant wear particle-induced osteolysis and inflammation in vivo. J Orthop Res. 34:58–64. 2016. View Article : Google Scholar : PubMed/NCBI
18	Keeney M, Waters H, Barcay K, Jiang X, Yao Z, Pajarinen J, Egashira K, Goodman SB and Yang F: Mutant MCP-1 protein delivery from layer-by-layer coatings on orthopedic implants to modulate inflammatory response. Biomaterials. 34:10287–10295. 2013. View Article : Google Scholar : PubMed/NCBI
19	Nabeshima A, Pajarinen J, Lin TH, Jiang X, Gibon E, Córdova LA, Loi F, Lu L, Jämsen E, Egashira K, et al: Mutant CCL2 protein coating mitigates wear particle-induced bone loss in a murine continuous polyethylene infusion model. Biomaterials. 117:1–9. 2017. View Article : Google Scholar : PubMed/NCBI
20	Severin IC, Souza AL, Davis JH, Musolino N, Mack M, Power CA and Proudfoot AE: Properties of 7ND-CCL2 are modulated upon fusion to Fc. Protein Eng Des Sel. 25:213–222. 2012. View Article : Google Scholar : PubMed/NCBI
21	Russmueller G, Moser D, Würger T, Wrba F, Christopoulos P, Kostakis G, Seemann R, Stadler V, Wimmer G, Kornek G, et al: Upregulation of osteoprotegerin expression correlates with bone invasion and predicts poor clinical outcome in oral cancer. Oral Oncol. 51:247–253. 2015. View Article : Google Scholar : PubMed/NCBI
22	Tada Y, Kokabu S, Sugiyama G, Nakatomi C, Aoki K, Fukushima H, Osawa K, Sugamori Y, Ohya K, Okamoto M, et al: The novel IκB kinase β inhibitor IMD-0560 prevents bone invasion by oral squamous cellcarcinoma. Oncotarget. 5:12317–12330. 2014. View Article : Google Scholar : PubMed/NCBI
23	Hsu FT, Chang B, Chen JC, Chiang IT, Liu YC, Kwang WK and Hwang JJ: Synergistic effect of sorafenib and radiation on human oral carcinoma in vivo. Sci Rep. 5:153912015. View Article : Google Scholar : PubMed/NCBI