Identification of brefelamide as a novel inhibitor of osteopontin that suppresses invasion of A549 lung cancer cells

Zhang,Jing; Yamada,Osamu; Kida,Shinya; Matsushita,Yoshihisa; Murase,Shinya; Hattori,Toshio; Kubohara,Yuzuru; Kikuchi,Haruhisa; Oshima,Yoshiteru

doi:10.3892/or.2016.5006

Identification of brefelamide as a novel inhibitor of osteopontin that suppresses invasion of A549 lung cancer cells

Authors:
- Jing Zhang
- Osamu Yamada
- Shinya Kida
- Yoshihisa Matsushita
- Shinya Murase
- Toshio Hattori
- Yuzuru Kubohara
- Haruhisa Kikuchi
- Yoshiteru Oshima
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Affiliations: Research and Development Center, FUSO Pharmaceutical Industries, Ltd., Osaka 536‑8523, Japan, Department of Disaster‑related Infectious Disease, International Research Institute of Disaster Science, Tohoku University, Sendai, Miyagi 980‑8575, Japan, Graduate School of Health and Sports Science, Juntendo University, Inzai, Chiba 270‑1695, Japan, Laboratory of Natural Product Chemistry, Graduate School of Pharmaceutical Sciences, Tohoku University, Sendai, Miyagi 980‑8578, Japan
Published online on: August 5, 2016 https://doi.org/10.3892/or.2016.5006
Pages: 2357-2364

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Abstract

The contribution of aberrant osteopontin (OPN) expression to tumor progression and metastasis has been documented in a wide spectrum of malignancies, and targeted inhibition of OPN has therefore emerged as an attractive strategy for cancer therapy. Transcription of OPN is regulated by various transcription factors, and our recently published study demonstrated that downregulation of OPN is an important event in the TGF‑β cytostatic program. We report here that brefelamide exerts an inhibitory effect on OPN expression and function in A549 human lung carcinoma cells. The promoter, RNA, and protein levels of OPN were decreased in brefelamide‑treated A549 cells, which was accompanied by reduced invasive ability in vitro. OPN inhibition by brefelamide was largely abrogated by disruption of a putative TGF‑β inhibitory element in the OPN promoter. Treatment with brefelamide induced Smad4 expression, and knockdown of Smad4 by RNA interference partially diminished the inhibitory effect of brefelamide on OPN. These results indicate that brefelamide inhibited OPN‑mediated cell invasion through restoration of the OPN repression by TGF‑β/Smad signaling. Together with the reported antiproliferative property, our findings suggest that brefelamide might serve as a potential candidate for the development of a new antitumor and antimetastatic agent.

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1	Weigelt B, Peterse JL and van't Veer LJ: Breast cancer metastasis: Markers and models. Nat Rev Cancer. 5:591–602. 2005. View Article : Google Scholar : PubMed/NCBI
2	Hanahan D and Weinberg RA: Hallmarks of cancer: The next generation. Cell. 144:646–674. 2011. View Article : Google Scholar : PubMed/NCBI
3	Nguyen DX, Bos PD and Massagué J: Metastasis: From dissemination to organ-specific colonization. Nat Rev Cancer. 9:274–284. 2009. View Article : Google Scholar : PubMed/NCBI
4	Joyce JA and Pollard JW: Microenvironmental regulation of metastasis. Nat Rev Cancer. 9:239–252. 2009. View Article : Google Scholar : PubMed/NCBI
5	Otranto M, Sarrazy V, Bonté F, Hinz B, Gabbiani G and Desmoulière A: The role of the myofibroblast in tumor stroma remodeling. Cell Adhes Migr. 6:203–219. 2012. View Article : Google Scholar
6	Mao Y, Keller ET, Garfield DH, Shen K and Wang J: Stromal cells in tumor microenvironment and breast cancer. Cancer Metastasis Rev. 32:303–315. 2013. View Article : Google Scholar
7	Geiger B, Bershadsky A, Pankov R and Yamada KM: Transmembrane crosstalk between the extracellular matrix - cytoskeleton crosstalk. Nat Rev Mol Cell Biol. 2:793–805. 2001. View Article : Google Scholar : PubMed/NCBI
8	Discher DE, Janmey P and Wang YL: Tissue cells feel and respond to the stiffness of their substrate. Science. 310:1139–1143. 2005. View Article : Google Scholar : PubMed/NCBI
9	Engler AJ, Sen S, Sweeney HL and Discher DE: Matrix elasticity directs stem cell lineage specification. Cell. 126:677–689. 2006. View Article : Google Scholar : PubMed/NCBI
10	Roberts DD: Emerging functions of matricellular proteins. Cell Mol Life Sci. 68:3133–3136. 2011. View Article : Google Scholar : PubMed/NCBI
11	Chiodoni C, Colombo MP and Sangaletti S: Matricellular proteins: From homeostasis to inflammation, cancer, and metastasis. Cancer Metastasis Rev. 29:295–307. 2010. View Article : Google Scholar : PubMed/NCBI
12	Brown LF, Berse B, Van de Water L, Papadopoulos-Sergiou A, Perruzzi CA, Manseau EJ, Dvorak HF and Senger DR: Expression and distribution of osteopontin in human tissues: Widespread association with luminal epithelial surfaces. Mol Biol Cell. 3:1169–1180. 1992. View Article : Google Scholar : PubMed/NCBI
13	Rangaswami H, Bulbule A and Kundu GC: Osteopontin: Role in cell signaling and cancer progression. Trends Cell Biol. 16:79–87. 2006. View Article : Google Scholar : PubMed/NCBI
14	Anborgh PH, Mutrie JC, Tuck AB and Chambers AF: Role of the metastasis-promoting protein osteopontin in the tumour microenvironment. J Cell Mol Med. 14:2037–2044. 2010. View Article : Google Scholar : PubMed/NCBI
15	Tuck AB, Arsenault DM, O'Malley FP, Hota C, Ling MC, Wilson SM and Chambers AF: Osteopontin induces increased invasiveness and plasminogen activator expression of human mammary epithelial cells. Oncogene. 18:4237–4246. 1999. View Article : Google Scholar : PubMed/NCBI
16	Lin YH and Yang-Yen HF: The osteopontin-CD44 survival signal involves activation of the phosphatidylinositol 3-kinase/Akt signaling pathway. J Biol Chem. 276:46024–46030. 2001. View Article : Google Scholar : PubMed/NCBI
17	Philip S, Bulbule A and Kundu GC: Osteopontin stimulates tumor growth and activation of promatrix metalloproteinase-2 through nuclear factor-kappa B-mediated induction of membrane type 1 matrix metalloproteinase in murine melanoma cells. J Biol Chem. 276:44926–44935. 2001. View Article : Google Scholar : PubMed/NCBI
18	Leali D, Dell'Era P, Stabile H, Sennino B, Chambers AF, Naldini A, Sozzani S, Nico B, Ribatti D and Presta M: Osteopontin (Eta-1) and fibroblast growth factor-2 cross-talk in angiogenesis. J Immunol. 171:1085–1093. 2003. View Article : Google Scholar : PubMed/NCBI
19	Furger KA, Menon RK, Tuck AB, Bramwell VH and Chambers AF: The functional and clinical roles of osteopontin in cancer and metastasis. Curr Mol Med. 1:621–632. 2001. View Article : Google Scholar
20	Kikuchi H, Saito Y, Sekiya J, Okano Y, Saito M, Nakahata N, Kubohara Y and Oshima Y: Isolation and synthesis of a new aromatic compound, brefelamide, from dictyostelium cellular slime molds and its inhibitory effect on the proliferation of astrocytoma cells. J Org Chem. 70:8854–8858. 2005. View Article : Google Scholar : PubMed/NCBI
21	Honma S, Saito M, Kikuchi H, Saito Y, Oshima Y, Nakahata N and Yoshida M: A reduction of epidermal growth factor receptor is involved in brefelamide-induced inhibition of phosphorylation of ERK in human astrocytoma cells. Eur J Pharmacol. 616:38–42. 2009. View Article : Google Scholar : PubMed/NCBI
22	Zhang J, Yamada O, Matsushita Y, Chagan-Yasutan H and Hattori T: Transactivation of human osteopontin promoter by human T-cell leukemia virus type 1-encoded Tax protein. Leuk Res. 34:763–768. 2010. View Article : Google Scholar
23	Zhang J, Yamada O, Kida S, Matsushita Y and Hattori T: Down-regulation of osteopontin mediates a novel mechanism underlying the cytostatic activity of TGF-β. Cell Oncol (Dordr). 39:119–128. 2016. View Article : Google Scholar
24	Zhang J, Yamada O, Sakamoto T, Yoshida H, Iwai T, Matsushita Y, Shimamura H, Araki H and Shimotohno K: Down-regulation of viral replication by adenoviral-mediated expression of siRNA against cellular cofactors for hepatitis C virus. Virology. 320:135–143. 2004. View Article : Google Scholar : PubMed/NCBI
25	Ding Z, Wu CJ, Chu GC, Xiao Y, Ho D, Zhang J, Perry SR, Labrot ES, Wu X, Lis R, et al: SMAD4-dependent barrier constrains prostate cancer growth and metastatic progression. Nature. 470:269–273. 2011. View Article : Google Scholar : PubMed/NCBI
26	Boissy P, Andersen TL, Abdallah BM, Kassem M, Plesner T and Delaissé JM: Resveratrol inhibits myeloma cell growth, prevents osteoclast formation, and promotes osteoblast differentiation. Cancer Res. 65:9943–9952. 2005. View Article : Google Scholar : PubMed/NCBI
27	Chakraborty G, Jain S, Patil TV and Kundu GC: Down-regulation of osteopontin attenuates breast tumour progression in vivo. J Cell Mol Med. 12(6A): 2305–2318. 2008. View Article : Google Scholar : PubMed/NCBI
28	Zhao J, Dong L, Lu B, Wu G, Xu D, Chen J, Li K, Tong X, Dai J, Yao S, et al: Down-regulation of osteopontin suppresses growth and metastasis of hepatocellular carcinoma via induction of apoptosis. Gastroenterology. 135:956–968. 2008. View Article : Google Scholar : PubMed/NCBI
29	Mi Z, Guo H, Russell MB, Liu Y, Sullenger BA and Kuo PC: RNA aptamer blockade of osteopontin inhibits growth and metastasis of MDA-MB231 breast cancer cells. Mol Ther. 17:153–161. 2009. View Article : Google Scholar
30	Hijiya N, Setoguchi M, Matsuura K, Higuchi Y, Akizuki S and Yamamoto S: Cloning and characterization of the human osteopontin gene and its promoter. Biochem J. 303:255–262. 1994. View Article : Google Scholar : PubMed/NCBI
31	Wang D, Yamamoto S, Hijiya N, Benveniste EN and Gladson CL: Transcriptional regulation of the human osteopontin promoter: Functional analysis and DNA-protein interactions. Oncogene. 19:5801–5809. 2000. View Article : Google Scholar : PubMed/NCBI
32	Kim HJ, Lee MH, Park HS, Park MH, Lee SW, Kim SY, Choi JY, Shin HI, Kim HJ and Ryoo HM: Erk pathway and activator protein 1 play crucial roles in FGF2-stimulated premature cranial suture closure. Dev Dyn. 227:335–346. 2003. View Article : Google Scholar : PubMed/NCBI
33	Joseph EW, Pratilas CA, Poulikakos PI, Tadi M, Wang W, Taylor BS, Halilovic E, Persaud Y, Xing F, Viale A, et al: The RAF inhibitor PLX4032 inhibits ERK signaling and tumor cell proliferation in a V600E BRAF-selective manner. Proc Natl Acad Sci USA. 107:14903–14908. 2010. View Article : Google Scholar : PubMed/NCBI
34	Demagny H, Araki T and De Robertis EM: The tumor suppressor Smad4/DPC4 is regulated by phosphorylations that integrate FGF, Wnt, and TGF-β signaling. Cell Reports. 9:688–700. 2014. View Article : Google Scholar
35	Alazzouzi H, Alhopuro P, Salovaara R, Sammalkorpi H, Järvinen H, Mecklin JP, Hemminki A, Schwartz S Jr, Aaltonen LA and Arango D: SMAD4 as a prognostic marker in colorectal cancer. Clin Cancer Res. 11:2606–2611. 2005. View Article : Google Scholar : PubMed/NCBI
36	Peng B, Fleming JB, Breslin T, Grau AM, Fojioka S, Abbruzzese JL, Evans DB, Ayers D, Wathen K, Wu T, et al: Suppression of tumorigenesis and induction of p15(ink4b) by Smad4/DPC4 in human pancreatic cancer cells. Clin Cancer Res. 8:3628–3638. 2002.PubMed/NCBI
37	Zhang B, Halder SK, Kashikar ND, Cho YJ, Datta A, Gorden DL and Datta PK: Antimetastatic role of Smad4 signaling in colorectal cancer. Gastroenterology. 138:969–980. 2010. View Article : Google Scholar
38	Tian X, Du H, Fu X, Li K, Li A and Zhang Y: Smad4 restoration leads to a suppression of Wnt/beta-catenin signaling activity and migration capacity in human colon carcinoma cells. Biochem Biophys Res Commun. 380:478–483. 2009. View Article : Google Scholar : PubMed/NCBI
39	Duda DG, Sunamura M, Lefter LP, Furukawa T, Yokoyama T, Yatsuoka T, Abe T, Inoue H, Motoi F, Egawa S, et al: Restoration of SMAD4 by gene therapy reverses the invasive phenotype in pancreatic adenocarcinoma cells. Oncogene. 22:6857–6864. 2003. View Article : Google Scholar : PubMed/NCBI
40	Tajima K, Ohashi R, Sekido Y, Hida T, Nara T, Hashimoto M, Iwakami S, Minakata K, Yae T, Takahashi F, et al: Osteopontin-mediated enhanced hyaluronan binding induces multidrug resistance in mesothelioma cells. Oncogene. 29:1941–1951. 2010. View Article : Google Scholar : PubMed/NCBI
41	Pang H, Cai L, Yang Y, Chen X, Sui G and Zhao C: Knockdown of osteopontin chemosensitizes MDA-MB-231 cells to cyclo-phosphamide by enhancing apoptosis through activating p38 MAPK pathway. Cancer Biother Radiopharm. 26:165–173. 2011. View Article : Google Scholar : PubMed/NCBI