Lumican mediates HTB94 chondrosarcoma cell growth via an IGF‑IR/Erk1/2 axis

Papoutsidakis,Antonis; Giatagana,Eirini   Maria; Berdiaki,Aikaterini; Spyridaki,Ioanna; Spandidos,Demetrios  A.; Tsatsakis,Aristidis; Tzanakakis,George N.; Nikitovic,Dragana

doi:10.3892/ijo.2020.5094

Lumican mediates HTB94 chondrosarcoma cell growth via an IGF‑IR/Erk1/2 axis

Authors:
- Antonis Papoutsidakis
- Eirini Maria Giatagana
- Aikaterini Berdiaki
- Ioanna Spyridaki
- Demetrios A. Spandidos
- Aristidis Tsatsakis
- George N. Tzanakakis
- Dragana Nikitovic
View Affiliations

Affiliations: Laboratory of Anatomy‑Histology‑Embryology, School of Medicine, University of Crete, 71003 Heraklion, Greece, Laboratory of Clinical Virology, School of Medicine, University of Crete, 71003 Heraklion, Greece, Laboratory of Toxicology, School of Medicine, University of Crete, 71003 Heraklion, Greece
Published online on: July 6, 2020 https://doi.org/10.3892/ijo.2020.5094
Pages: 791-803

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

Chondrosarcoma is a malignant bone tumor characterized by the production of a modified cartilage‑type extracellular matrix (ECM). In the present study, the expression levels of the small leucine‑rich proteoglycans (SLRPs), decorin, biglycan and lumican, were examined in the HTB94 human chondrosarcoma cell line. HTB94 cells were found to express and secrete the 3 SLRP members. RT‑qPCR and western blot analysis demonstrated that lumican was the most abundantly secreted SLRP, whereas decorin and biglycan expression levels were low. The utilization of short interfering RNA specific for the decorin, biglycan, and lumican genes resulted in the efficient downregulation of the respective mRNA levels (P≤0.001). The growth of the HTB94 cells was stimulated by lumican (P≤0.001), whereas their migration and adhesion were not affected (P=NS). By contrast, these cellular functions were not sensitive to a decrease in low endogenous levels of decorin and biglycan. Lumicandeficiency significantly inhibited both basal and insulin‑like growth factor I (IGF‑I)‑induced HTB94 cell growth (P≤0.001 andP≤0.01, respectively). These effects were executed through the insulin‑like growth factor I receptor (IGF‑IR), whose activation was markedly attenuated (P≤0.01) in lumican‑deficient HTB94 cells. The downregulation of lumican induced the substantial inhibition of extracellular regulated kinase (ERK1/2) activation (P≤ 0.01), indicating that ERK1/2 is a necessary component of lumican/IGF‑IR‑mediated HTB94 cell proliferation. Moreover, the lumican‑deficient cells exhibit increased mRNA levels of p53 (P≤0.05), suggesting that lumican facilitates HTB94 cell growth through an IGF‑IR/ERK1/2/p53 signaling cascade. On the whole, the findings of the present study demonstrate that endogenous lumican is a novel regulator of HTB94 cell growth.

View Figures

View References

1	International Agency for Research on Cancer (IARC): WHO Classification of Tumours of Soft Tissue and Bone. Fletcher CDM, Bridge JA, Hogendoorn PCW and Mertens F: 5. 4th edition. IARC; Lyon: 2013
2	Heck RK Jr, Peabody TD and Simon MA: Staging of primary malignancies of bone. CA Cancer J Clin. 56:366–375. 2006. View Article : Google Scholar : PubMed/NCBI
3	Chow WA: Update on chondrosarcomas. Curr Opin Oncol. 19:371–376. 2007. View Article : Google Scholar : PubMed/NCBI
4	Tsuda Y, Ogura K, Hakozaki M, Kikuta K, Ae K, Tsuchiya H, Iwata S, Ueda T, Kawano H and Kawai A: Mesenchymal chon-drosarcoma: A Japanese Musculoskeletal Oncology Group (JMOG) study on 57 patients. J Surg Oncol. 115:760–767. 2017. View Article : Google Scholar : PubMed/NCBI
5	Theocharis AD, Manou D and Karamanos NK: The extracellular matrix as a multitasking player in disease. FEBS J. 286:2830–2869. 2019. View Article : Google Scholar : PubMed/NCBI
6	Nikitovic D, Papoutsidakis A, Karamanos NK and Tzanakakis GN: Lumican affects tumor cell functions, tumor-ECM interactions, angiogenesis and inflammatory response. Matrix Biol. 35:206–214. 2014. View Article : Google Scholar
7	Varol C and Sagi I: Phagocyte-extracellular matrix crosstalk empowers tumor development and dissemination. FEBS J. 285:734–751. 2018. View Article : Google Scholar
8	Yuzhalin AE, Lim SY, Kutikhin AG and Gordon-Weeks AN: Dynamic matrisome: ECM remodeling factors licensing cancer progression and metastasis. Biochim Biophys Acta Rev Cancer. 1870:207–228. 2018. View Article : Google Scholar : PubMed/NCBI
9	Kessenbrock K, Plaks V and Werb Z: Matrix metalloproteinases: Regulators of the tumor microenvironment. Cell. 141:52–67. 2010. View Article : Google Scholar : PubMed/NCBI
10	Neill T, Schaefer L and Iozzo RV: Decoding the Matrix: Instructive Roles of Proteoglycan Receptors. Biochemistry. 54:4583–4598. 2015. View Article : Google Scholar : PubMed/NCBI
11	Karamanou K, Perrot G, Maquart FX and Brézillon S: Lumican as a multivalent effector in wound healing. Adv Drug Deliv Rev. 129:344–351. 2018. View Article : Google Scholar : PubMed/NCBI
12	Nikitovic D, Aggelidakis J, Young MF, Iozzo RV, Karamanos NK and Tzanakakis GN: The biology of small leucine-rich proteoglycans in bone pathophysiology. J Biol Chem. 287:33926–33933. 2012. View Article : Google Scholar : PubMed/NCBI
13	Chen S and Birk DE: The regulatory roles of small leucine-rich proteoglycans in extracellular matrix assembly. FEBS J. 280:2120–2137. 2013. View Article : Google Scholar : PubMed/NCBI
14	Schaefer L and Iozzo RV: Biological functions of the small leucine-rich proteoglycans: From genetics to signal transduction. J Biol Chem. 283:21305–21309. 2008. View Article : Google Scholar : PubMed/NCBI
15	Merline R, Schaefer RM and Schaefer L: The matricellular functions of small leucine-rich proteoglycans (SLRPs). J Cell Commun Signal. 3:323–335. 2009. View Article : Google Scholar : PubMed/NCBI
16	Grogan SP, Chen X, Sovani S, Taniguchi N, Colwell CW Jr, Lotz MK and D'Lima DD: Influence of cartilage extracellular matrix molecules on cell phenotype and neocartilage formation. Tissue Eng Part A. 20:264–274. 2014. View Article : Google Scholar :
17	Lewis JL, Krawczak DA, Oegema TR Jr and Westendorf JJ: Effect of decorin and dermatan sulfate on the mechanical properties of a neocartilage. Connect Tissue Res. 51:159–170. 2010. View Article : Google Scholar
18	Burdan F, Szumiło J, Korobowicz A, Farooquee R, Patel S, Patel A, Dave A, Szumiło M, Solecki M, Klepacz R, et al: Morphology and physiology of the epiphyseal growth plate. Folia Histochem Cytobiol. 47:5–16. 2009. View Article : Google Scholar : PubMed/NCBI
19	Melrose J, Shu C, Whitelock JM and Lord MS: The cartilage extracellular matrix as a transient developmental scaffold for growth plate maturation. Matrix Biol. 52-54:363–383. 2016. View Article : Google Scholar : PubMed/NCBI
20	Appunni S, Anand V, Khandelwal M, Gupta N, Rubens M and Sharma A: Small Leucine Rich Proteoglycans (decorin, biglycan and lumican) in cancer. Clin Chim Acta. 491:1–7. 2019. View Article : Google Scholar : PubMed/NCBI
21	Xu L, Li Z, Liu SY, Xu SY and Ni GX: Asporin and osteoarthritis. Osteoarthritis Cartilage. 23:933–939. 2015. View Article : Google Scholar : PubMed/NCBI
22	Ni GX, Li Z and Zhou YZ: The role of small leucine-rich proteoglycans in osteoarthritis pathogenesis. Osteoarthritis Cartilage. 22:896–903. 2014. View Article : Google Scholar : PubMed/NCBI
23	Söderström M, Böhling T, Ekfors T, Nelimarkka L, Aro HT and Vuorio E: Molecular profiling of human chondrosarcomas for matrix production and cancer markers. Int J Cancer. 100:144–151. 2002. View Article : Google Scholar : PubMed/NCBI
24	Chansky H, Robbins JR, Cha S, Raskind WH, Conrad EU and Sandell LJ: Expression of cartilage extracellular matrix and potential regulatory genes in a new human chondrosarcoma cell line. J Orthop Res. 16:521–530. 1998. View Article : Google Scholar : PubMed/NCBI
25	Nikitovic D, Chalkiadaki G, Berdiaki A, Aggelidakis J, Katonis P, Karamanos NK and Tzanakakis GN: Lumican regulates osteosarcoma cell adhesion by modulating TGFβ2 activity. Int J Biochem Cell Biol. 43:928–935. 2011. View Article : Google Scholar : PubMed/NCBI
26	Berdiaki A, Datsis G, Nikitovic D, Tsatsakis A, Katonis P, Karamanos N and Tzanakakis G: Parathyroid hormone (PTH) peptides through the regulation of hyaluronan metabolism affect osteosarcoma cell migration. IUBMB Life. 62:377–386. 2010.PubMed/NCBI
27	Voudouri K, Nikitovic D, Berdiaki A, Kletsas D, Karamanos NK and Tzanakakis GN: IGF-I/EGF and E2 signaling crosstalk through IGF-IR conduit point affects breast cancer cell adhesion. Matrix Biol. 56:95–113. 2016. View Article : Google Scholar : PubMed/NCBI
28	Im GI, Jung NH and Tae SK: Chondrogenic differentiation of mesenchymal stem cells isolated from patients in late adulthood: The optimal conditions of growth factors. Tissue Eng. 12:527–536. 2006. View Article : Google Scholar : PubMed/NCBI
29	Khaghani SAB, Akbarova G, Soon CF and Dilbazi G: Effect of transforming growth factor-β2 on biological regulation of multilayer primary chondrocyte culture. Cell Tissue Bank. 19:763–775. 2018. View Article : Google Scholar : PubMed/NCBI
30	Boumédiene K, Takigawa M and Pujol JP: Cell density-dependent proliferative effects of transforming growth factor (TGF)-β 1, β 2, and β 3 in human chondrosarcoma cells HCS-2/8 are associated with changes in the expression of TGF-β receptor type I. Cancer Invest. 19:475–486. 2001. View Article : Google Scholar
31	Boeuf S, Bovée JV, Lehner B, van den Akker B, van Ruler M, Cleton-Jansen AM and Richter W: BMP and TGFbeta pathways in human central chondrosarcoma: Enhanced endoglin and Smad 1 signaling in high grade tumors. BMC Cancer. 12:4882012. View Article : Google Scholar : PubMed/NCBI
32	De Luca F: Regulatory Role for Growth Hormone in Statural Growth: IGF-Dependent and IGF-Independent Effects on Growth Plate Chondrogenesis and Longitudinal Bone Growth. Pediatr Endocrinol Rev. 16(Suppl 1): 33–38. 2018.PubMed/NCBI
33	Hiraoka K, Zenmyo M, Komiya S, Kawabata R, Yokouchi M, Suzuki R, Hamada T, Kato S and Nagata K: Relationship of p21 (waf1/cip1) and differentiation in chondrosarcoma cells. Virchows Arch. 440:285–290. 2002. View Article : Google Scholar : PubMed/NCBI
34	Mytilinaiou M, Nikitovic D, Berdiaki A, Papoutsidakis A, Papachristou DJ, Tsatsakis A and Tzanakakis GN: IGF-I regulates HT1080 fibrosarcoma cell migration through a syndecan-2/Erk/ezrin signaling axis. Exp Cell Res. 361:9–18. 2017. View Article : Google Scholar : PubMed/NCBI
35	Nikitovic D, Berdiaki A, Zafiropoulos A, Katonis P, Tsatsakis A, Karamanos NK and Tzanakakis GN: Lumican expression is positively correlated with the differentiation and negatively with the growth of human osteosarcoma cells. FEBS J. 275:350–361. 2008. View Article : Google Scholar
36	Coulson-Thomas VJ, Coulson-Thomas YM, Gesteira TF, Andrade de Paula CA, Carneiro CR, Ortiz V, Toma L, Kao WW and Nader HB: Lumican expression, localization and antitumor activity in prostate cancer. Exp Cell Res. 319:967–981. 2013. View Article : Google Scholar : PubMed/NCBI
37	Mao W, Luo M, Huang X, Wang Q, Fan J, Gao L, Zhang Y and Geng J: Knockdown of lumican inhibits proliferation and migration of bladder cancer. Transl Oncol. 12:1072–1078. 2019. View Article : Google Scholar : PubMed/NCBI
38	Chen L, Zhang Y, Zuo Y, Ma F and Song H: Lumican expression in gastric cancer and its association with biological behavior and prognosis. Oncol Lett. 14:5235–5240. 2017.PubMed/NCBI
39	Hsiao KC, Chu PY, Chang GC and Liu KJ: Elevated Expression of Lumican in Lung Cancer Cells Promotes Bone Metastasis through an Autocrine Regulatory Mechanism. Cancers (Basel). 12:2332020. View Article : Google Scholar
40	Yang CT, Hsu PC and Chow SE: Downregulation of lumican enhanced mitotic defects and aneuploidy in lung cancer cells. Cell Cycle. 19:97–108. 2020. View Article : Google Scholar
41	Chen X, Li X, Hu X, Jiang F, Shen Y, Xu R, Wu L, Wei P and Shen X: LUM Expression and Its Prognostic Significance in Gastric Cancer. Front Oncol. 10:6052020. View Article : Google Scholar : PubMed/NCBI
42	Karamanou K, Franchi M, Piperigkou Z, Perreau C, Maquart FX, Vynios DH and Brézillon S: Lumican effectively regulates the estrogen receptors-associated functional properties of breast cancer cells, expression of matrix effectors and epithelial-to-mesenchymal transition. Sci Rep. 7:451382017. View Article : Google Scholar : PubMed/NCBI
43	Jeanne A, Untereiner V, Perreau C, Proult I, Gobinet C, Boulagnon-Rombi C, Terryn C, Martiny L, Brézillon S and Dedieu S: Lumican delays melanoma growth in mice and drives tumor molecular assembly as well as response to matrix-targeted TAX2 therapeutic peptide. Sci Rep. 7:77002017. View Article : Google Scholar : PubMed/NCBI
44	Rahimi RA and Leof EB: TGF-β signaling: A tale of two responses. J Cell Biochem. 102:593–608. 2007. View Article : Google Scholar : PubMed/NCBI
45	Herpin A and Cunningham C: Cross-talk between the bone morphogenetic protein pathway and other major signaling pathways results in tightly regulated cell-specific outcomes. FEBS J. 274:2977–2985. 2007. View Article : Google Scholar : PubMed/NCBI
46	Iozzo RV, Buraschi S, Genua M, Xu SQ, Solomides CC, Peiper SC, Gomella LG, Owens RC and Morrione A: Decorin antagonizes IGF receptor I (IGF-IR) function by interfering with IGF-IR activity and attenuating downstream signaling. J Biol Chem. 286:34712–34721. 2011. View Article : Google Scholar : PubMed/NCBI
47	Baker J, Liu JP, Robertson EJ and Efstratiadis A: Role of insulin-like growth factors in embryonic and postnatal growth. Cell. 75:73–82. 1993. View Article : Google Scholar : PubMed/NCBI
48	Liu JP, Baker J, Perkins AS, Robertson EJ and Efstratiadis A: Mice carrying null mutations of the genes encoding insulin-like growth factor I (Igf-1) and type 1 IGF receptor (Igf1r). Cell. 75:59–72. 1993.PubMed/NCBI
49	Vlahovic G, Meadows KL, Hatch AJ, Jia J, Nixon AB, Uronis HE, Morse MA, Selim MA, Crawford J, Riedel RF, et al: A Phase I Trial of the IGF-1R Antibody Ganitumab (AMG 479) in Combination with Everolimus (RAD001) and Panitumumab in Patients with Advanced Cancer. Oncologist. 23:782–790. 2018. View Article : Google Scholar : PubMed/NCBI
50	Peterse EF, Cleven AH, De Jong Y, Briaire-de Bruijn I, Fletcher JA, Danen EH, Cleton-Jansen AM and Bovée JV: No preclinical rationale for IGF1R directed therapy in chondro-sarcoma of bone. BMC Cancer. 16:4752016. View Article : Google Scholar
51	Schönherr E, Sunderkötter C, Iozzo RV and Schaefer L: Decorin, a novel player in the insulin-like growth factor system. J Biol Chem. 280:15767–15772. 2005. View Article : Google Scholar : PubMed/NCBI
52	Aggelidakis J, Berdiaki A, Nikitovic D, Papoutsidakis A, Papachristou DJ, Tsatsakis AM and Tzanakakis GN: Biglycan regulates MG63 osteosarcoma cell growth through a LPR6/ β-catenin/IGFR-IR signaling axis. Front Oncol. 8:4702018. View Article : Google Scholar
53	Matsumura T, Whelan MC, Li XQ and Trippel SB: Regulation by IGF-I and TGF-beta1 of Swarm-rat chondrosarcoma chon-drocytes. J Orthop Res. 18:351–355. 2000. View Article : Google Scholar : PubMed/NCBI
54	Metalli D, Lovat F, Tripodi F, Genua M, Xu SQ, Spinelli M, Alberghina L, Vanoni M, Baffa R, Gomella LG, et al: The insulin-like growth factor receptor I promotes motility and invasion of bladder cancer cells through Akt- and mitogen-activated protein kinase-dependent activation of paxillin. Am J Pathol. 176:2997–3006. 2010. View Article : Google Scholar : PubMed/NCBI
55	Cantini G, Lombardi A, Piscitelli E, Poli G, Ceni E, Marchiani S, Ercolino T, Galli A, Serio M, Mannelli M, et al: Rosiglitazone inhibits adrenocortical cancer cell proliferation by interfering with the IGF-IR intracellular signaling. PPAR Res. 2008:9040412008. View Article : Google Scholar : PubMed/NCBI
56	Qin J, Shaukat I, Mainard D, Netter P, Barré L and Ouzzine M: Constitutive activation of EGFR is associated with tumor progression and plays a prominent role in malignant phenotype of chondrosarcoma. Oncotarget. 10:3166–3182. 2019. View Article : Google Scholar : PubMed/NCBI
57	Kamemura N, Murakami S, Komatsu H, Sawanoi M, Miyamoto K, Ishidoh K, Kishimoto K, Tsuji A and Yuasa K: Type II cGMP-dependent protein kinase negatively regulates fibroblast growth factor signaling by phosphorylating Raf-1 at serine 43 in rat chondrosarcoma cells. Biochem Biophys Res Commun. 483:82–87. 2017. View Article : Google Scholar : PubMed/NCBI
58	Guan PP, Yu X, Guo JJ, Wang Y, Wang T, Li JY, Konstantopoulos K, Wang ZY and Wang P: By activating matrix metalloproteinase-7, shear stress promotes chondrosarcoma cell motility, invasion and lung colonization. Oncotarget. 6:9140–9159. 2015. View Article : Google Scholar : PubMed/NCBI
59	Lee KS, Kim SW and Lee HS: Orostachys japonicus induce p53-dependent cell cycle arrest through the MAPK signaling pathway in OVCAR-3 human ovarian cancer cells. Food Sci Nutr. 6:2395–2401. 2018. View Article : Google Scholar : PubMed/NCBI
60	Boroumand Moghaddam A, Moniri M, Azizi S, Abdul Rahim R, Bin Ariff A, Navaderi M and Mohamad R: Eco-Friendly Formulated Zinc Oxide Nanoparticles: Induction of Cell Cycle Arrest and Apoptosis in the MCF-7 Cancer Cell Line. Genes (Basel). 8:2812017. View Article : Google Scholar
61	Dai X, Ma W, He X and Jha RK: Review of therapeutic strategies for osteosarcoma, chondrosarcoma, and Ewing's sarcoma. Med Sci Monit. 17:RA177–RA190. 2011. View Article : Google Scholar : PubMed/NCBI
62	Mery B, Espenel S, Guy JB, Rancoule C, Vallard A, Aloy MT, Rodriguez-Lafrasse C and Magné N: Biological aspects of chon-drosarcoma: Leaps and hurdles. Crit Rev Oncol Hematol. 126:32–36. 2018. View Article : Google Scholar : PubMed/NCBI
63	Philippou A, Christopoulos PF and Koutsilieris DM: Clinical studies in humans targeting the various components of the IGF system show lack of efficacy in the treatment of cancer. Mutat Res Rev Mutat Res. 772:105–122. 2017. View Article : Google Scholar : PubMed/NCBI
64	Fiedler LR, Schönherr E, Waddington R, Niland S, Seidler DG, Aeschlimann D and Eble JA: Decorin regulates endothelial cell motility on collagen I through activation of insulin-like growth factor I receptor and modulation of α2β1 integrin activity. J Biol Chem. 283:17406–17415. 2008. View Article : Google Scholar : PubMed/NCBI
65	Santra M, Eichstetter I and Iozzo RV: An anti-oncogenic role for decorin. Down-regulation of ErbB2 leads to growth suppression and cytodifferentiation of mammary carcinoma cells. J Biol Chem. 275:35153–35161. 2000. View Article : Google Scholar : PubMed/NCBI
66	Zhang W, Ge Y, Cheng Q, Zhang Q, Fang L and Zheng J: Decorin is a pivotal effector in the extracellular matrix and tumour micro-environment. Oncotarget. 9:5480–5491. 2018. View Article : Google Scholar : PubMed/NCBI
67	Xing X, Gu X and Ma T: Knockdown of biglycan expression by RNA interference inhibits the proliferation and invasion of, and induces apoptosis in, the HCT116 colon cancer cell line. Mol Med Rep. 12:7538–7544. 2015. View Article : Google Scholar : PubMed/NCBI
68	Schaefer L, Tredup C, Gubbiotti MA and Iozzo RV: Proteoglycan neofunctions: Regulation of inflammation and autophagy in cancer biology. FEBS J. 284:10–26. 2017. View Article : Google Scholar :
69	Liu B, Xu T, Xu X, Cui Y and Xing X: Biglycan promotes the chemotherapy resistance of colon cancer by activating NF-κB signal transduction. Mol Cell Biochem. 449:285–294. 2018. View Article : Google Scholar : PubMed/NCBI
70	Schulz GB, Grimm T, Sers C, Riemer P, Elmasry M, Kirchner T, Stief CG, Karl A and Horst D: Prognostic value and association with epithelial-mesenchymal transition and molecular subtypes of the proteoglycan biglycan in advanced bladder cancer. Urol Oncol. 37:530.e9–530.e18. 2019. View Article : Google Scholar