Osteopontin inhibits HIF-2α mRNA expression in osteoarthritic chondrocytes

Cheng,Chao; Zhang,Fang‑Jie; Tian,Jian; Tu,Min; Xiong,Yi‑Lin; Luo,Wei; Li,Yu‑Sheng; Song,Bing‑Bing; Gao,Shu‑Guang; Lei,Guang‑Hua

doi:10.3892/etm.2015.2434

Osteopontin inhibits HIF-2α mRNA expression in osteoarthritic chondrocytes

Authors:
- Chao Cheng
- Fang‑Jie Zhang
- Jian Tian
- Min Tu
- Yi‑Lin Xiong
- Wei Luo
- Yu‑Sheng Li
- Bing‑Bing Song
- Shu‑Guang Gao
- Guang‑Hua Lei
View Affiliations

Affiliations: Department of Orthopaedics, Xiangya Hospital, Central South University, Changsha, Hunan, 410008, P.R. China, Hunan Province Environmental Monitoring Center, Changsha, Hunan 410019, P.R. China
Published online on: April 20, 2015 https://doi.org/10.3892/etm.2015.2434
Pages: 2415-2419

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 aim of the present study was to investigate the in vitro effect of osteopontin (OPN) on the expression of hypoxia‑inducible factor‑2α (HIF‑2α) in chondrocytes and the role of OPN in osteoarthritis (OA). Cartilage was purified from the tibial surfaces of patients with OA of the knee and cultured in vitro to obtain chondrocytes. Recombinant human OPN (rhOPN) and OPN small interfering RNA (siRNA) were used to treat the chondrocytes, and the changes in the expression levels of the HIF‑2α gene were measured. An anti‑CD44 blocking monoclonal antibody (mAb) was used to determine the probable ligand‑receptor interactions. Reverse transcription‑quantitative polymerase chain reaction assays were designed and validated with SYBR® Green dyes for the simultaneous quantiﬁcation of the mRNA expression levels of OPN and HIF‑2α. The mRNA expression level of HIF‑2α was markedly decreased in the rhOPN‑treated group compared with that in the control group; by contrast, OPN siRNA increased HIF‑2α gene expression. CD44 blocking mAb suppressed the inhibitory effect of OPN on HIF‑2α mRNA expression. The results of the present study suggest that OPN may play a protective role in OA by inhibiting HIF‑2α gene expression in osteoarthritic chondrocytes through CD44 interaction.

View Figures

View References

1	Gravallese EM: Osteopontin: A bridge between bone and the immune system. J Clin Invest. 112:147–149. 2003. View Article : Google Scholar : PubMed/NCBI
2	Zhang FJ, Gao SG, Cheng L, et al: The effect of hyaluronic acid on osteopontin and CD44 mRNA of fibroblast-like synoviocytes in patients with osteoarthritis of the knee. Rheumatol Int. 33:79–83. 2013. View Article : Google Scholar : PubMed/NCBI
3	Denhardt DT and Noda M: Osteopontin expression and function: Role in bone remodeling. J Cell Biochem Suppl. 30–31:92–102. 1998. View Article : Google Scholar
4	Lampe MA, Patarca R, Iregui MV and Cantor H: Polyclonal B cell activation by the Eta-1 cytokine and the development of systemic autoimmune disease. J Immunol. 147:2902–2906. 1991.PubMed/NCBI
5	Pullig O, Weseloh G, Gauer S and Swoboda B: Osteopontin is expressed by adult human osteoarthritic chondrocytes: Protein and mRNA analysis of normal and osteoarthritic cartilage. Matrix Biol. 19:245–255. 2000. View Article : Google Scholar : PubMed/NCBI
6	Honsawek S, Tanavalee A, Sakdinakiattikoon M, Chayanupatkul M and Yuktanandana P: Correlation of plasma and synovial fluid osteopontin with disease severity in knee osteoarthritis. Clin Biochem. 42:808–812. 2009. View Article : Google Scholar : PubMed/NCBI
7	Gao SG, Li KH, Zeng KB, Tu M, Xu M and Lei GH: Elevated osteopontin level of synovial fluid and articular cartilage is associated with disease severity in knee osteoarthritis patients. Osteoarthritis Cartilage. 18:82–87. 2010. View Article : Google Scholar : PubMed/NCBI
8	Weber GF, Ashkar S, Glimcher MJ and Cantor H: Receptor-ligand interaction between CD44 and osteopontin (Eta-1). Science. 271:509–512. 1996. View Article : Google Scholar : PubMed/NCBI
9	Dunn S, Kolomytkin OV, Waddell DD and Marino AA: Hyaluronan-binding receptors: Possible involvement in osteoarthritis. Mod Rheumatol. 19:151–155. 2009. View Article : Google Scholar : PubMed/NCBI
10	Zhang FJ, Luo W, Gao SG, et al: Expression of CD44 in articular cartilage is associated with disease severity in knee osteoarthritis. Mod Rheumatol. 23:1186–1191. 2013. View Article : Google Scholar : PubMed/NCBI
11	Milner PI, Fairfax TP, Browning JA, Wilkins RJ and Gibson JS: The effect of O2 tension on pH homeostasis in equine articular chondrocytes. Arthritis Rheum. 54:3523–3532. 2006. View Article : Google Scholar : PubMed/NCBI
12	Semenza GL: HIF-1 and human disease: One highly involved factor. Genes Dev. 14:1983–1991. 2000.PubMed/NCBI
13	Semenza GL: Hypoxia-inducible factors in physiology and medicine. Cell. 148:399–408. 2012. View Article : Google Scholar : PubMed/NCBI
14	Smith TG, Robbins PA and Ratcliffe PJ: The human side of hypoxia-inducible factor. Br J Haematol. 141:325–334. 2008. View Article : Google Scholar : PubMed/NCBI
15	Saito T, Fukai A, Mabuchi A, et al: Transcriptional regulation of endochondral ossification by HIF-2alpha during skeletal growth and osteoarthritis development. Nat Med. 16:678–686. 2010. View Article : Google Scholar : PubMed/NCBI
16	Yang S, Kim J, Ryu JH, et al: Hypoxia-inducible factor-2alpha is a catabolic regulator of osteoarthritic cartilage destruction. Nat Med. 16:687–693. 2010. View Article : Google Scholar : PubMed/NCBI
17	Attur MG, Dave MN, Stuchin S, et al: Osteopontin: An intrinsic inhibitor of inflammation in cartilage. Arthritis Rheum. 44:578–584. 2001. View Article : Google Scholar : PubMed/NCBI
18	Matsui Y, Iwasaki N, Kon S, et al: Accelerated development of aging-associated and instability-induced osteoarthritis in osteopontin-deficient mice. Arthritis Rheum. 60:2362–2371. 2009. View Article : Google Scholar : PubMed/NCBI
19	Wilhelm J and Pingoud A: Real-time polymerase chain reaction. ChemBioChem. 4:1120–1128. 2003. View Article : Google Scholar : PubMed/NCBI
20	Ponchel F, Toomes C, Bransfield K, et al: Real-time PCR based on SYBR-Green I fluorescence: An alternative to the TaqMan assay for a relative quantification of gene rearrangements, gene amplifications and micro gene deletions. BMC Biotechnol. 3:182003. View Article : Google Scholar : PubMed/NCBI
21	Martens G, Cai Y, Hinke S, Stangé G, Van de Casteele M and Pipeleers D: Nutrient sensing in pancreatic beta cells suppresses mitochondrial superoxide generation and its contribution to apoptosis. Biochem Soc Trans. 33:300–301. 2005. View Article : Google Scholar : PubMed/NCBI
22	Sandell LJ and Aigner T: Articular cartilage and changes in arthritis. An introduction: Cell biology of osteoarthritis. Arthritis Res. 3:107–13. 2001. View Article : Google Scholar : PubMed/NCBI
23	Goldring MB and Goldring SR: Osteoarthritis. J Cell Physiol. 213:626–34. 2007. View Article : Google Scholar : PubMed/NCBI
24	Kühn K, D'Lima DD, Hashimoto S and Lotz M: Cell death in cartilage. Osteoarthritis Cartilage. 12:1–16. 2004. View Article : Google Scholar : PubMed/NCBI
25	Hashimoto S, Ochs RL, Komiya S and Lotz M: Linkage of chondrocyte apoptosis and cartilage degradation in human osteoarthritis. Arthritis Rheum. 41:1632–1638. 1998. View Article : Google Scholar : PubMed/NCBI
26	Zamli Z and Sharif M: Chondrocyte apoptosis: A cause or consequence of osteoarthritis? Int J Rheum Dis. 14:159–166. 2011. View Article : Google Scholar : PubMed/NCBI
27	Ryu JH, Shin Y, Huh YH, Yang S, Chun CH and Chun JS: Hypoxia-inducible factor-2 α regulates Fas-mediated chondrocyte apoptosis during osteoarthritic cartilage destruction. Cell Death Differ. 19:440–450. 2012. View Article : Google Scholar : PubMed/NCBI
28	Zhu Y, Denhardt DT, Cao H, et al: Hypoxia upregulates osteopontin expression in NIH-3T3 cells via a Ras-activated enhancer. Oncogene. 24:6555–6563. 2005.PubMed/NCBI
29	Underhill C: CD44: The hyaluronan receptor. J Cell Sci. 103:293–298. 1992.PubMed/NCBI
30	Ishii S, Ford R, Thomas P, Nachman A, Steele G Jr and Jessup JM: CD44 participates in the adhesion of human colorectal carcinoma cells to laminin and type IV collagen. Surg Oncol. 2:255–264. 1993. View Article : Google Scholar : PubMed/NCBI
31	Isacke CM: The role of the cytoplasmic domain in regulating CD44 function. J Cell Sci. 107:2353–2359. 1994.PubMed/NCBI
32	Knudson CB, Nofal GA, Pamintuan L and Aguiar DJ: The chondrocyte pericellular matrix: A model for hyaluronan-mediated cell-matrix interactions. Biochem Soc Trans. 27:142–147. 1999.PubMed/NCBI