Objective: To assess the modulating function of osteopontin (OPN) on human Tenon's fibroblasts (HTFs). Methods: With the intervention of OPN in different concentrations (0, 0.05 μmol/L, 0.5 μmol/L,and 5 μmol/L), the proliferation rate of HTFs was measured by 3-(4,5-dimethylthiazol-2-Yl)-2,5-
diphenyltetrazolium bromide (MTT), and the motility was surveyed by the cell scratch method. Real-time polymerase chain reaction (PCR) was applied to measure the mRNA of vascular endothelial growth factor(VEGF) and transforming growth factor β (TGF-β) in HTFs in the presence of different concentration of OPN. Western blot was used to measure the collagen-I levels. Data were analyzed using ANOVA. Results:OPN induced the proliferation of HTFs as indicated by MTT and enhanced the motility of HTFs as shownby the cell scratch method. The effects were concentration dependent (F =174.5, 297.1, P <0.01). In the presence of anti-OPN 1A12 (5 μmol/L OPN+20 μg/ml 1A12), the effects of OPN on HTFs were inhibited. None of the concentrations of OPN affected mRNA levels of VEGF and TGF-β mRNA. OPN augmented HTF expression of collagen-I protein in a concentration-dependent manner (F =30.2, P <0.01). Conclusions:The enhancement of HTF proliferation, motility, and collagen-I expression OPN was concentration dependent. Incubation with the monoclonal OPN antibody 1A12 inhibited the effects of OPN on the HTFs.
李维娜,程金伟,魏锐利. 骨桥蛋白及其单克隆抗体1A12对人Tenon's囊成纤维细胞的调控作用[J]. 中华眼视光学与视觉科学杂志, 2017, 19(6): 329-334.
Weina Li, Jinwei Cheng, Ruili Wei.. The Modulating Function of Osteopontin and Osteopontin Monoclonal Antibody in Human Tenon's Fibroblasts. Chinese Journal of Optometry Ophthalmology and Visual science, 2017, 19(6): 329-334. DOI: DOI:10.3760/cma.j.issn.1674-845X.2017.06.003
Senger DR, Wirth DF, Hynes RO. Transformed mammalian cells secrete specific proteins and phosphoproteins. Cell, 1979, 16(4): 885-893.
[2]
Weber GF. The metastasis gene osteopontin: a candidate target for cancer therapy. Biochim Biophys Acta, 2001, 1552(2): 61-85.
[3]
Wang KX, Denhardt DT. Osteopontin: role in immune regulation and stress responses. Cytokine Growth Factor Rev, 2008, 19(5-6): 333-345. DOI: 10.1016/j.cytogfr.2008.08.001.
[4]
Zohar R, Zhu B, Liu P, et al. Increased cell death in osteopontin-deficient cardiac fibroblasts occurs by a caspase-3-independent pathway. Am J Physiol Heart Circ Physiol, 2004, 287(4): H1730-1739. DOI: 10.1152/ajpheart.00098.2004.
[5]
He B, Mirza M, Weber GF. An osteopontin splice variant induces anchorage independence in human breast cancer cells. Oncogene, 2006, 25(15): 2192-2202. DOI: 10.1038/sj.onc.1209248.
[6]
Khan SA, Lopez-Chua CA, Zhang J, et al. Soluble osteopontin inhibits apoptosis of adherent endothelial cells deprived of growth factors. J Cell Biochem, 2002, 85(4): 728-736. DOI: 10.1002/jcb.10170.
[7]
Urtasun R, Lopategi A, George J, et al. Osteopontin, an oxidant stress sensitive cytokine, up-regulates collagen-I via integrin α(V)β(3) engagement and PI3K/pAkt/NFκB signaling.Hepatology, 2012, 55(2): 594-608. DOI: 10.1002/hep.24701.
[8]
Tan TK, Zheng G, Hsu TT, et al. Matrix metalloproteinase-9 of tubular and macrophage origin contributes to the pathogenesis of renalfibrosis viamacrophagerecruitment throughosteopontin cleavage. Lab Invest, 2013, 93(4): 434-449. DOI: 10.1038/ labinvest.2013.3.
[9]
Tsukui T, Ueha S, Abe J, et al. Qualitative rather than quantitative changes are hallmarks of fibroblasts in bleomycin-induced pulmonary fibrosis. Am J Pathol, 2013, 183(3): 7 58-773. DOI: 10.1016/j.ajpath.2013.06.005.
[10]
Klingel K, Kandolf R. Osteopontin: a biomarker to predict the outcome of inflammatory heart disease. Semin Thromb Hemost, 2010, 36(2): 195-202. DOI: 10.1055/s-0030-1251504.
[11]
Wu M, Schneider DJ, Mayes MD, et al. Osteopontin in systemic sclerosis and its role in dermal fibrosis. J Invest Dermatol, 2012, 132(6): 1605-1614. DOI: 10.1038/jid.2012.32.
[12]
Hunter C, Bond J, Kuo PC, et al. The role of osteopontin and osteopontinaptamer (OPN-R3) in fibroblast activity. J Surg Res, 2012, 176(1): 348-358. DOI: 10.1016/j.jss.2011.07.054.
[13]
Kohan M, Breuer R, Berkman N. Osteopontin induces airway remodeling and lung fibroblast activation in a murine model of asthma. Am J Respir Cell Mol Biol, 2009, 41(3): 290-296. DOI: 10.1165/rcmb.2008-0307OC.
[14]
O'NeillEC,QinQ,VanBergenNJ,etal.Antifibroticactivityof bevacizumab on human Tenon's fibroblasts in vitro. Invest Ophthalmol Vis Sci, 2010, 51(12): 6524-6532. DOI: 10.1167/iovs. 10-5669.
[15]
Cordeiro MF, Bhattacharya SS, Schultz GS, et al. TGF-beta1, -beta2, and-beta3invitro:biphasiceffectsonTenon'sfibroblastcontraction, proliferation, and migration. Invest Ophthalmol Vis Sci, 2000, 41(3): 756-763.
[16]
Gupta A, Zhou CQ, Chellaiah MA. Osteopontin and MMP9: Associations with VEGF Expression/Secretion and Angiogenesis in PC3 Prostate Cancer Cells. Cancers (Basel), 2013, 5(2): 617-638. DOI: 10.3390/cancers5020617.
[17]
Sun J, Feng A, Chen S, et al. Osteopontin splice variants expressed by breast tumors regulate monocyte activation via MCP-1 and