TSitologiya i Genetika 2023, vol. 57, no. 3, 57-59
Cytology and Genetics 2023, vol. 57, no. 3, 282–289, doi: https://www.doi.org/10.3103/S0095452723030027

TGF-β1 transactivates ADAMTS-2 (ADAM Metallopeptidase With Thrombospondin Type 1 Motif 2) in Saos-2 Cells through canonical and non-canonical pathways

Alper M.

ul>
  • Department of Translational Oncology Institute of Oncology, Dokuz Eylül University, 35330 Balçova, İzmir/Turkey
  • Osteosarcoma is a malignant bone tumor that is common in children and adolescents. The tumor microenvironment is highly effective in the development and progression of osteosarcoma. Transforming growth factor-β (TGF-β) is one of the most abundant cytokines in the tumor microenvironment, and can regulate tumor initiation, progression, and metastasis promoting extracellular matrix (ECM) remodeling and epithelial-mesenchymal transition (EMT). ADAMTS (ADAM Metallopeptidase With Thrombospondin Motifs) proteases have critical functions in normal and tumor microenvironments by processing individual proteins in the ECM. ADAMTSs contribute to tissue remodeling, inflammation, cell migration and, angiogenesis. Among the family members, ADAMTS-2 is a well-known example for ECM remodeling which cleaves the N-terminal propeptide of procollagen and promotes correct collagen fibrillogenesis. Cytokines can regulate normal and tumor microenvironments by affecting ECM proteins. In this study, the effect of TGF-β1, on the transcriptional regulation of the ADAMTS-2, which is an essential enzyme for ECM remodeling was investigated in Saos-2 cells. TGF-β1 upregulated ADAMTS-2 expression both at mRNA and protein levels. Transient transfection assays revealed that TGF-β1 was also induced ADAMTS-2 promoter activity. According to the pathway inhibition studies, both canonical and non-canonical signaling pathways and post-translational mechanisms were responsible for the induction. These studies will contribute to future research on ADAMTS-2 mediated ECM remodeling in osteosarcoma.

    Keywords: ADAMTS-2, TGF-β1, Osteosarcoma, Saos-2, Transcriptional Regulation

    TSitologiya i Genetika
    2023, vol. 57, no. 3, 57-59

    Current Issue
    Cytology and Genetics
    2023, vol. 57, no. 3, 282–289,
    doi: 10.3103/S0095452723030027

    Full text and supplemented materials

    References

    Alper, M. and Kockar, F., IL-6 upregulates a disintegrin and metalloproteinase with thrombospondin motifs 2 (ADAMTS-2) in human osteosarcoma cells mediated by JNK pathway, Mol. Cell. Biochem., 2014, vol. 393, nos. 1–2, pp. 165–175. https://doi.org/10.1007/s11010-014-2056-9

    Alper, M., Aydemir, A.T., and Köçkar, F., Induction of human ADAMTS-2 gene expression by IL-1α is mediated by a multiple crosstalk of MEK/JNK and PI3K pathways in osteoblast like cells, Gene, 2015, vol. 573, no. 2, pp. 321–327. https://doi.org/10.1016/j.gene.2015.07.064

    Berridge, M.V. and Tan, A.S., Characterization of the cellular reduction of 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT): subcellular localization, substrate dependence, and involvement of mitochondrial electron transport in MTT reduction, Arch. Biochem. Biophys., 1993, vol. 303, no. 2, pp. 474–482. https://doi.org/10.1006/abbi.1993.1311

    Clayton, S.W., Ban, G.I., Liu, C., et al., Canonical and noncanonical TGF-β signaling regulate fibrous tissue differentiation in the axial skeleton, Sci. Rep., 2020, vol. 10, p. 21364.https://doi.org/10.1038/s41598-020-78206-4

    Corre, I., Verrecchia, F., Crenn, vol., et al., The osteosarcoma microenvironment: a complex but targetable ecosystem, Cells, 2020, vol. 9, no. 4, p. 976. https://doi.org/10.3390/cells9040976

    Cui, J., Dean, D., Hornicek, F.J., et al., The role of extracellular matrix in osteosarcoma progression and metastasis, J. Exp. Clin. Cancer Res., 2020, vol. 39, no. 1, p. 178. https://doi.org/10.1186/s13046-020-01685-w

    Czekanska, E.M., Stoddart, M.J., Richards, R.G., et al., In search of an osteoblast cell model for in vitro research, Eur. Cells Mater., 2012, vol. 24, pp. 1–17. https://doi.org/10.22203/ecm.v024a01

    Dong, F., Liu, T., Jin, H., et al., Chimaphilin inhibits human osteosarcoma cell invasion and metastasis through suppressing the TGF-β1-induced epithelial-to-mesenchymal transition markers via PI-3K/Akt, ERK1/2, and Smad signaling pathways, Can. J. Physiol. Pharmacol., 2018, vol. 96, no. 1, pp. 1–7. https://doi.org/10.1139/cjpp-2016-0522

    El Mabrouk, M., Sylvester, J., and Zafarullah, M., Signaling pathways implicated in oncostatin M-Induced aggrecanase-1 and matrix metalloproteinase-13 expression in human articular chondrocytes, Biochim. Biophys. Acta, Mol. Cell Res., 2007, vol. 1773, pp. 309–320. https://doi.org/10.1016/j.bbamcr.2006.11.018

    Girden, E.R., ANOVA: Repeated Measures, Sage, 1992.

    Hubmacher, D. and Apte, S.S., ADAMTS proteins as modulators of microfibril formation and function, Matrix Biol., 2015, vol. 47, pp. 34–43. https://doi.org/10.1016/j.matbio.2015.05.004

    Jiang, C., Zhou, Y., Huang, Y., et al., Overexpression of ADAMTS-2 in tumor cells and stroma is predictive of poor clinical prognosis in gastric cancer, Hum. Pathol., 2019, vol. 84, pp. 44–51. https://doi.org/10.1016/j.humpath.2018.08.030

    Kelwick, R., Desanlis, I., Wheeler, G.N., et al., The ADAMTS (A Disintegrin and Metalloproteinase with Thrombospondin motifs) family, Genome Biol., 2015, vol. 16, no. 1, p. 113. https://doi.org/10.1186/s13059-015-0676-3

    Kockar, F.T., Foka, P., Hughes, T.R., et al., Analysis of the Xenopus laevis CCAAT-enhancer binding protein α gene promoter demonstrates species-specific differences in the mechanisms for both autoactivation and regulation by Sp1, Nucleic Acids Res., 2001, vol. 29, pp. 362–372. https://doi.org/10.1093/nar/29.2.362

    Li, F., Li, S., and Cheng, T., TGF-β1 Promotes Osteosarcoma Cell Migration and Invasion Through the miR-143-Versican Pathway, Cell Physiol. Biochem., 2014, vol. 34, pp. 2169–2179. https://doi.org/10.1159/000369660

    Liu, S., Chen, S., and Zeng, J., TGF-β signaling: A complex role in tumorigenesis, Mol. Med. Rep., 2018, vol. 17, no. 1, pp. 699–704. https://doi.org/10.3892/mmr.2017.7970

    Livak, K.J. and Schmittgen, T.D., Analysis of relative gene expression data using real-time quantitative PCR and the 2−ΔΔC T Method, Methods, 2001, vol. 25, no. 4, pp. 402–408. https://doi.org/10.1006/meth.2001.1262

    Lopes‑Júnior, L.C., Silveira, D.S.C., Vulczak, A., et al., Emerging cytokine networks in osteosarcoma, Oncol. Commun., 2016, vol. 2, p. e1167. https://doi.org/10.14800/CCM.1510

    Mimata, Y., Kamataki, A., Oikawa S., et al., Interleukin-6 Upregulates Expression of ADAMTS-4 in Fibroblast-like Synoviocytes from Patients with Rheumatoid Arthritis, Int. J. Rheum. Dis., 2012, vol. 15, pp. 36–44. https://doi.org/10.1111/j.1756-185X.2011.01656.x

    Minitab L.L.C. 2021. Minitab. Retrieved from https://www.minitab.com.

    Pautke, C., Schieker, M., Tischer, T., et al., Characterization of osteosarcoma cell lines MG-63, Saos-2 and U‑2 OS in comparison to human osteoblasts, Anticancer Res., 2004, vol. 24, no. 6, pp. 3743–3748.

    Poos, K., Smida, J., Maugg, D., et al., Genomic heterogeneity of osteosarcoma- shift from single candidates to functional modules, PLoS One, 2015, vol. 10, no. 4, p. e0123082. https://doi.org/10.1371/journal.pone.0123082

    Rose, K.W.J., Taye, N., Karoulias, S.Z., et al., Regulation of ADAMTS proteases, Front. Mol. Biosci., 2021, vol. 29, no. 8, p. 701959. https://doi.org/10.3389/fmolb.2021.701959

    Satz-Jacobowitz, B. and Hubmacher, D., The quest for substrates and binding partners: A critical barrier for understanding the role of ADAMTS proteases in musculoskeletal development and disease, Dev. Dyn., 2021, vol. 250, no. 1, pp. 8–26. https://doi.org/10.1002/dvdy.248

    Schneider, C.A., Rasband, W.S., and Eliceiri, K.W., NIH image to image: 25 years of image analysis, Nat. Methods, 2021, vol. 9, pp. 671–675. https://doi.org/10.1016/j.lfs.2015.12.010

    Schnellmann, R., Sack, R., Hess, D., et al., A selective extracellular matrix proteomics approach identifies fibronectin proteolysis by a disintegrin-like and metalloprotease domain with thrombospondin type 1 motifs (ADAMTS16) and its impact on spheroid morphogenesis, Mol. Cell. Proteomics, 2018, vol. 17, pp. 1410–1425. https://doi.org/10.1074/mcp.RA118.000676

    Surridge, A.K., Rodgers, U.R., Swingler, T.E., et al., Characterization and regulation of ADAMTS-16, Matrix B-iol., 2009, vol. 28, pp. 416–424. https://doi.org/10.1016/j.matbio.2009.07.001

    Takeda, S., Three-dimensional domain architecture of the ADAM family proteinases. Semin, Cell Dev. Biol., 2009, vol. 20, pp. 146–152. https://doi.org/10.1016/j.semcdb.2008.07.009

    Takeuchi, Y., Fukumoto, S., and Matsumoto, T., Relationship between actions of transforming growth factor (TGF)-beta and cell surface expression of its receptors in clonal osteoblastic cells, J. Cell. Physiol., 1995, vol. 162, no. 3, pp. 315–321. https://doi.org/10.1002/jcp.1041620303

    Théret, N., Bouezzedine, F., Azar, F., et al., ADAM and ADAMTS proteins, new players in the regulation of hepatocellular carcinoma microenvironment, Cancers (Basel), 2021, vol. 13, no. 7, p.1563. https://doi.org/10.3390/cancers13071563

    Tokay, E. and Kockar, F., Identification of intracellular pathways through which TGF β1 upregulates URG-4/URGCP gene expression in hepatoma cells, Life Sci., 2016, vol. 144, pp. 121–128. https://doi.org/10.1016/j.lfs.2015.12.010

    Tokay, E., Sagkan, R.I., and Kockar, F., TNF-α Induces URG-4/URGCP gene expression in hepatoma cells through starvation dependent manner, Biochem. Ge-net., 2021, vol. 59, pp. 300–314 https://doi.org/10.1007/s10528-020-09972-z

    Uchida, K., Takano, S., Matsumoto, T., et al., transforming growth factor activating kinase 1 regulates extracellular matrix degrading enzymes and pain-related molecule expression following tumor necrosis factor-α a stimulation of synovial cells: an in vitro study, BMC Musculoskeletal Disord., 2017, vol. 18, p. 283. https://doi.org/10.1186/s12891-017-1648-4

    Verrecchia, F. and Rédini, F., Transforming Growth factor-β signaling plays a pivotal role in the interplay between osteosarcoma cells and their microenvironment, Front. Oncol., 2018, vol. 30, no. 8, p. 133. https://doi.org/10.3389/fonc.2018.00133

    Wang, W.M., Lee, S., Steiglitz, B.M., et al., Transforming growth factor beta induces secretion of activated ADAMTS-2. A procollagen III N-proteinase, J. Biol. Chem., 2003, vol. 278, pp. 19549–19557. https://doi.org/10.1074/jbc.M300767200

    Xiao, H., Chen, L., Luo, G., et al., Effect of the cytokine levels in serum on osteosarcoma, Tumor Biol., 2014, vol. 35, no. 2, pp. 1023–1028. https://doi.org/10.1007/s13277-013-1136-x

    Zhang, Y, Alexander, P.B., and Wang, X.F., TGF-β Family Signaling in the Control of Cell Proliferation and Survival, Cold Spring Harbor Perspect. Biol., 2017, vol. 9, no. 4, p. a022145. https://doi.org/10.1101/cshperspect.a022145