Knockdown of macrophage inhibitory cytokine-1 in RPMI-8226 human multiple myeloma cells inhibits osteoclastic differentiation through inhibiting the RANKL-Erk1/2 signaling pathway

Yuan,Mingzhou; Chen,Junmin; Zeng,Zhiyong

doi:10.3892/mmr.2016.5879

Knockdown of macrophage inhibitory cytokine-1 in RPMI-8226 human multiple myeloma cells inhibits osteoclastic differentiation through inhibiting the RANKL-Erk1/2 signaling pathway

Authors:
- Mingzhou Yuan
- Junmin Chen
- Zhiyong Zeng
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Affiliations: Department of Hematology and Rheumatology, The First Clinical Medical College of Fujian Medical University, Fuzhou, Fujian 350005, P.R. China
Published online on: October 24, 2016 https://doi.org/10.3892/mmr.2016.5879
Pages: 5199-5204

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Abstract

Patients with multiple myeloma (MM) often develop myeloma bone disease (MBD). The development of MBD from MM is considered to be caused by an abnormal bone marrow microenvironment. Macrophage inhibitory cytokine-1 (MIC-1) is a member of the transforming growth factor‑β superfamily. In patients with MM, MIC‑1 is expressed at high levels, however, whether this increased expression of MIC‑1 is associated with the development of MBD from MM remains to be elucidated. The present study investigated whether MIC‑1 is essential for the osteoclastic differentiation of peripheral blood mononuclear cells (PBMNCs) by using a co‑culture system, in which the PBMNCs were co‑cultured with RPMI‑8226 cells. The expression of MIC‑1 in the RPMI‑8226 cells was knocked down using RNA interference. Osteoclastic differentiation was evaluated using tartrate‑resistant acid phosphatase staining and lacunar resorption on dentine slices. The expression of receptor activator of nuclear factor‑κB ligand (RANKL) and phosphorylation of extracellular signal‑regulated kinase (Erk)1/2 were measured using Western blotting. It was found that the reduced expression of MIC‑1 in the RPMI‑8226 cells inhibited the osteoclastic differentiation of PBMNCs and decreased the expression levels of RANKL and phosphorylated Erk1/2. It was concluded that MIC‑1 promoted the osteoclastic differentiation of PBMNCs via the RANKL‑Erk1/2 signaling pathway and, therefore, MIC‑1 may offer potential as a target in the design of strategies to treat MBD.

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1	Kyle RA and Rajkumar SV: Criteria for diagnosis, staging, risk stratification and response assessment of multiple myeloma. Leukemia. 23:3–9. 2009. View Article : Google Scholar : PubMed/NCBI
2	Corre J, Mahtouk K, Attal M, Gadelorge M, Huynh A, Fleury-Cappellesso S, Danho C, Laharrague P, Klein B, Rème T and Bourin P: Bone marrow mesenchymal stem cells are abnormal in multiple myeloma. Leukemia. 21:1079–1088. 2007.PubMed/NCBI
3	Hromas R, Hufford M, Sutton J, Xu D, Li Y and Lu L: PLAB, a novel placental bone morphogenetic protein. Biochim Biophys Acta. 1354:40–44. 1997. View Article : Google Scholar : PubMed/NCBI
4	Paralkar VM, Vail AL, Grasser WA, Brown TA, Xu H, Vukicevic S, Ke HZ, Qi H, Owen TA and Thompson DD: Cloning and characterization of a novel member of the transforming growth factor-beta/bone morphogenetic protein family. J Biol Chem. 273:13760–13767. 1998. View Article : Google Scholar : PubMed/NCBI
5	Böttner M, Suter-Crazzolara C, Schober A and Unsicker K: Expression of a novel member of the TGF-beta superfamily, growth/differentiation factor-15/macrophage-inhibiting cytokine-1 (GDF-15/MIC-1) in adult rat tissues. Cell Tissue Res. 297:103–110. 1999. View Article : Google Scholar : PubMed/NCBI
6	Baek SJ, Horowitz JM and Eling TE: Molecular cloning and characterization of human nonsteroidal anti-inflammatory drug-activated gene promoter. Basal transcription is mediated by Sp1 and Sp3. J Biol Chem. 276:33384–33392. 2001. View Article : Google Scholar : PubMed/NCBI
7	Bootcov MR, Bauskin AR, Valenzuela SM, Moore AG, Bansal M, He XY, Zhang HP, Donnellan M, Mahler S, Pryor K, et al: Mic-1, a novel macrophage inhibitory cytokine, is a divergent member of the TGF-beta superfamily. Proc Natl Acad Sci USA. 94:11514–11519. 1997. View Article : Google Scholar : PubMed/NCBI
8	Tarkun P, Atesoglu E Birtas, Mehtap O, Musul MM and Hacihanefioglu A: Serum growth differentiation factor 15 levels in newly diagnosed multiple myeloma patients. Acta Haematol. 131:173–178. 2014. View Article : Google Scholar : PubMed/NCBI
9	Wakchoure S, Swain TM, Hentunen TA, Bauskin AR, Brown DA, Breit SN, Vuopala KS, Harris KW and Selander KS: Expression of macrophage inhibitory cytokine-1 in prostate cancer bone metastases induces osteoclast activation and weight loss. Prostate. 69:652–661. 2009. View Article : Google Scholar : PubMed/NCBI
10	Zeng Z, Zhang C and Chen J: Lentivirus-mediated RNA interference of DC-STAMP expression inhibits the fusion and resorptive activity of human osteoclasts. J Bone Miner Metab. 31:409–416. 2013. View Article : Google Scholar : PubMed/NCBI
11	Livak KJ and Schmittgen TD: Analysis of relative gene expression data using real-time quantitative PCR and the 2(−Delta Delta C(T)) Method. Methods. 25:402–408. 2001. View Article : Google Scholar : PubMed/NCBI
12	Roodman GD: Pathogenesis of myeloma bone disease. Leukemia. 23:435–441. 2009. View Article : Google Scholar : PubMed/NCBI
13	Khosla S: Minireview: The OPG/RANKL/RANK system. Endocrinology. 142:5050–5055. 2001. View Article : Google Scholar : PubMed/NCBI
14	Sung B, Prasad S, Yadav VR, Gupta SC, Reuter S, Yamamoto N, Murakami A and Aggarwal BB: RANKL signaling and osteoclastogenesis is negatively regulated by cardamonin. PLoS One. 8:e641182013. View Article : Google Scholar : PubMed/NCBI
15	Lacey DL, Timms E, Tan HL, Kelley MJ, Dunstan CR, Burgess T, Elliott R, Colombero A, Elliott G, Scully S, et al: Osteoprotegerin ligand is a cytokine that regulates osteoclast differentiation and activation. Cell. 93:165–176. 1998. View Article : Google Scholar : PubMed/NCBI
16	Kong YY, Yoshida H, Sarosi I, Tan HL, Timms E, Capparelli C, Morony S, Oliveira-dos-Santos AJ, Van G, Itie A, et al: OPGL is a key regulator of osteoclastogenesis, lymphocyte development and lymph-node organogenesis. Nature. 397:315–323. 1999. View Article : Google Scholar : PubMed/NCBI
17	Schramek D, Leibbrandt A, Sigl V, Kenner L, Pospisilik JA, Lee HJ, Hanada R, Joshi PA, Aliprantis A, Glimcher L, et al: Osteoclast differentiation factor RANKL controls development of progestin-driven mammary cancer. Nature. 468:98–102. 2010. View Article : Google Scholar : PubMed/NCBI
18	Sung B, Oyajobi B and Aggarwal BB: Plumbagin inhibits osteoclastogenesis and reduces human breast cancer-induced osteolytic bone metastasis in mice through suppression of RANKL signaling. Mol Cancer Ther. 11:350–359. 2012. View Article : Google Scholar : PubMed/NCBI
19	Ai LS, Sun CY, Zhang L, Zhou SC, Chu ZB, Qin Y, Wang YD, Zeng W, Yan H, Guo T, et al: Inhibition of BDNF in multiple myeloma blocks osteoclastogenesis via down-regulated stroma-derived RANKL expression both in vitro and in vivo. PLoS One. 7:e462872012. View Article : Google Scholar : PubMed/NCBI
20	Wada T, Nakashima T, Hiroshi N and Penninger JM: RANKL-RANK signaling in osteoclastogenesis and bone disease. Trends Mol Med. 12:17–25. 2006. View Article : Google Scholar : PubMed/NCBI
21	Huang P, Han J and Hui L: MAPK signaling in inflammation-associated cancer development. Protein Cell. 1:218–226. 2010. View Article : Google Scholar : PubMed/NCBI
22	Zhang W and Liu HT: MAPK signal pathways in the regulation of cell proliferation in mammalian cells. Cell Res. 12:9–18. 2002. View Article : Google Scholar : PubMed/NCBI
23	Feng X: RANKing intracellular signaling in osteoclasts. IUBMB Life. 57:389–395. 2005. View Article : Google Scholar : PubMed/NCBI