Mycobacterium tuberculosis 10-kDa co-chaperonin regulates the expression levels of receptor activator of nuclear factor-κB ligand and osteoprotegerin in human osteoblasts

Zhang,Yuanyu; Liu,Xia; Li,Kun; Bai,Jingping

doi:10.3892/etm.2014.2153

Mycobacterium tuberculosis 10-kDa co-chaperonin regulates the expression levels of receptor activator of nuclear factor-κB ligand and osteoprotegerin in human osteoblasts

Authors:
- Yuanyu Zhang
- Xia Liu
- Kun Li
- Jingping Bai
View Affiliations

Affiliations: Department of Orthopedics, Affiliated Tumor Hospital of Xinjiang Medical University, Urumqi, Xinjiang 830000, P.R. China, Department of Pathology, First Affiliated Hospital of Xinjiang Medical University, Urumqi, Xinjiang 830000, P.R. China
Published online on: December 22, 2014 https://doi.org/10.3892/etm.2014.2153
Pages: 919-924

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 effect of recombinant Mycobacterium tuberculosis (r‑Mt) 10‑kDa co‑chaperonin (cpn10) on the expression of osteoprotegerin (OPG) and receptor activator of nuclear factor‑κB ligand (RANKL) in third‑generation cultured osteoblasts. The osteoblast‑like cultures were isolated from bone fragments taken from patients undergoing surgery. Prior to stimulation with r‑Mt cpn10, cells were incubated in serum‑free medium for 24 h. r‑Mt cpn10 was added into fresh serum‑free medium, reaching final concentrations of 0.01‑10 µg/ml. The levels of OPG were determined using enzyme‑linked immunosorbent assay. Reverse transcription‑quantitative polymerase chain reaction (RT‑qPCR) analysis was performed to determine the levels of RANKL and OPG mRNA. For measurement of the protein levels of OPG and RANKL, a western blotting assay was performed. r‑Mt cpn10 downregulated the protein levels of OPG in the third generation cultured osteoblasts at a dose of 10 µg/ml. RT‑qPCR revealed that the OPG mRNA level was decreased by 73% after 4 h and by 85.5% after 8 h following incubation with r‑Mt cpn10 (10 µg/ml). Western blot analysis demonstrated similar results for the OPG protein level. In the third‑generation cultured osteoblasts, the levels of RANKL mRNA and protein were increased by 2.6‑ and 1‑fold, respectively, following incubation with r‑Mt cpn10 (10 µg/ml). Furthermore, the RANKL/OPG ratio was markedly increased by r‑Mt cpn10 (10 µg/ml) treatment. In conclusion, the results of the current study demonstrated that r‑Mt cpn10 decreased the levels of OPG and increased the levels of RANKL in a dose‑ and time‑dependent manner. Notably, the present study indicated that r‑Mt cpn10 exerts its effect on osteoblastic cells by increasing the RANKL/OPG ratio.

View Figures

View References

1	Andersen TL, Sondergaard TE, Skorzynska KE, et al: A physical mechanism for coupling bone resorption and formation in adult human bone. Am J Pathol. 174:239–247. 2009. View Article : Google Scholar :
2	Trouvin AP and Goëb V: Receptor activator of nuclear factor-κB ligand and osteoprotegrin: maintaining the balance to prevent bone loss. Clin Interv Aging. 5:345–354. 2010.
3	Dougall WC: Molecular pathways: osteoclast-dependent and osteoclast-independent roles of the RANKL/RANK/OPG pathway in tumorigenesis and metastasis. Clin Cancer Res. 18:326–335. 2012. View Article : Google Scholar
4	Raju R, Balakrishnan L, Nanjappa V, et al: A comprehensive manually curated reaction map of RANKL/RANK-signalling pathway. Database (Oxford). 2011:bar0212011. View Article : Google Scholar
5	Singh PP, van der Kraan AG, Xu J, Gillespie MT and Quinn JM: Membrane-bound receptor activator of NFκB ligand (RANKL) activity displayed by osteoblasts is differentially regulated by osteolytic factors. Biochem Biophys Res Commun. 422:48–53. 2012. View Article : Google Scholar : PubMed/NCBI
6	Rachner TD, Khosla S and Hofbauer LC: Osteoporosis: now and the future. Lancet. 377:1276–1287. 2011. View Article : Google Scholar : PubMed/NCBI
7	Wu WT, Lee RP, Wang CH, et al: The association of serum osteoprotegerin and osteoporosis in postmenopausal hemodialysis patients: a pilot study. J Womens Health (Larchmt). 19:785–790. 2010. View Article : Google Scholar
8	Reitmanova S and Gustafson DL: Coloring the white plague: a syndemic approach to immigrant tuberculosis in Canada. Ethn Health. 17:403–418. 2012. View Article : Google Scholar
9	Glaziou P, Falzon D, Floyd K and Raviglione M: Global epidemiology of tuberculosis. Semin Respir Crit Care Med. 34:3–16. 2013. View Article : Google Scholar : PubMed/NCBI
10	Corrado A, Neve A, Macchiarola A, et al: RANKL/OPG ratio and DKK-1 expression in primary osteoblastic cultures from osteoarthritic and osteoporotic subjects. J Rheumatol. 40:684–694. 2013. View Article : Google Scholar : PubMed/NCBI
11	Henderson B, Lund PA and Coates AR: Multiple moonlighting functions of mycobacterial molecular chaperons. Tuberculosis (Edinb). 90:119–124. 2010. View Article : Google Scholar
12	Taneja B and Mande SC: Metal ions modulate the plastic nature of Mycobacterium tuberculosis chaperonin-10. Protein Eng. 14:391–395. 2001. View Article : Google Scholar : PubMed/NCBI
13	Meghji S, White PA, Nair SP, et al: Mycobacterium tuberculosis chaperonin 10 stimulates bone resorption: a potential contributory factor in Pott’s disease. J Exp Med. 186:1241–1246. 1997. View Article : Google Scholar : PubMed/NCBI
14	Jonsson KB, Frost A, Nilsson O, Ljunghall S and Ljunggren O: Three isolation techniques for primary culture of human osteoblastlike cells: a comparison. Acta Orthop Scand. 70:365–373. 1999. View Article : Google Scholar : PubMed/NCBI
15	Brändström H, Björkman T and Ljunggren O: Regulation of osteoprotegerin secretion from primary cultures of human bone marrow stromal cells. Biochem Biophys Res Commun. 280:831–835. 2001. View Article : Google Scholar : PubMed/NCBI
16	Suda T, Takahashi N, Udagawa N, et al: Modulation of osteoclast differentiation and function by the new members of the tumor necrosis factor receptor and ligand families. Endocr Rev. 20:345–357. 1999. View Article : Google Scholar : PubMed/NCBI
17	Morony S, Tintut Y, Zhang Z, et al: Osteoprotegerin inhibits vascular calcification without affecting atherosclerosis in ldlr(−/−) mice. Circulation. 117:411–420. 2008. View Article : Google Scholar : PubMed/NCBI
18	Jones DH, Nakashima T, Sanchez OH, et al: Regulation of cancer cell migration and bone metastasis by RANKL. Nature. 440:692–696. 2006. View Article : Google Scholar : PubMed/NCBI
19	Wang Y, Yang C, Xie WL, et al: Puerarin concurrently stimulates osteoprotegerin and inhibits receptor activator of NF-κB ligand (RANKL) and interleukin-6 production in human osteoblastic MG-63 cells. Phytomedicine. 21:1032–1036. 2014. View Article : Google Scholar : PubMed/NCBI
20	Xiong Q, Zhang LC, Zhang LH, et al: Effects of recombinant human osteoprotegerin and recombinant RANK protein on the differentiation of osteoclast precursors. Zhongguo Gu Shang. 26:324–327. 2013.PubMed/NCBI
21	Goranova-Marinova V, Goranov S, Pavlov P and Tzvetkova T: Serum levels of OPG, RANKL and RANKL/OPG ratio in newly-diagnosed patients with multiple myeloma. Clinical correlations. Haematologica. 92:1000–1001. 2007. View Article : Google Scholar : PubMed/NCBI
22	Samelson EJ, Broe KE, Demissie S, et al: Increased plasma osteoprotegerin concentrations are associated with indices of bone strength of the hip. J Clin Endocrinol Metab. 93:1789–1795. 2008. View Article : Google Scholar : PubMed/NCBI
23	Kearns AE, Khosla S and Kostenuik PJ: Receptor activator of nuclear factor kappaB ligand and osteoprotegerin regulation of bone remodeling in health and disease. Endocr Rev. 29:155–192. 2008. View Article : Google Scholar
24	Lamoureux F, Richard P, Wittrant Y, et al: Therapeutic relevance of osteoprotegerin gene therapy in osteosarcoma: blockade of the vicious cycle between tumor cell proliferation and bone resorption. Cancer Res. 167:7308–7318. 2007. View Article : Google Scholar
25	Joseph F, Chan BY, Durham BH, et al: The circadian rhythm of osteoprotegerin and its association with parathyroid hormone secretion. J Clin Endocrinol Metab. 92:3230–3238. 2007. View Article : Google Scholar : PubMed/NCBI
26	Roberts MM, Coker AR, Fossati G, et al: Mycobacerium tuberculosis chaperonin 10 heptamers self-associate through their biologically active loops. J Bacteriol. 185:4172–4185. 2003. View Article : Google Scholar : PubMed/NCBI
27	Qamra R, Mande SC, Coates AR and Henderson B: The unusual chaperonins of Mycobacterium tuberculosis. Tuberculosis (Edinb). 85:385–394. 2005. View Article : Google Scholar
28	Roberts MM, Coker AR, Fossati G, et al: Mycobacterium tuberculosis chaperonin 10 heptamers self-associate through their biologically active loops. J Bacteriol. 185:4172–4185. 2003. View Article : Google Scholar : PubMed/NCBI
29	Castanié-Cornet MP, Bruel N and Genevaux P: Chaperone networking facilitates protein targeting to the bacterial cytoplasmic membrane. Biochim Biophys Acta. 1843:1442–1456. 2014. View Article : Google Scholar
30	Henderson B, Lund PA and Coates AR: Multiple moonlighting functions of mycobacterial molecular chaperones. Tuberculosis (Edinb). 90:119–124. 2010. View Article : Google Scholar
31	Bai YD, Yang FS, Xuan K, Bai YX and Wu BL: Inhibition of RANK/RANKL signal transduction pathway: a promising approach for osteoporosis treatment. Med Hypotheses. 71:256–258. 2008. View Article : Google Scholar : PubMed/NCBI