Oridonin inhibits the proliferation of human osteosarcoma cells by suppressing Wnt/β-catenin signaling

Liu,Yang; Liu,Ying-Zi; Zhang,Ran-Xi; Wang,Xing; Meng,Zi-Jun; Huang,Jun; Wu,Ke; Luo,Jin-Yong; Zuo,Guo-Wei; Chen,Liang; Yin,Liang-Jun; Deng,Zhong-Liang; He,Bai-Cheng

doi:10.3892/ijo.2014.2456

Oridonin inhibits the proliferation of human osteosarcoma cells by suppressing Wnt/β-catenin signaling

Authors:
- Yang Liu
- Ying-Zi Liu
- Ran-Xi Zhang
- Xing Wang
- Zi-Jun Meng
- Jun Huang
- Ke Wu
- Jin-Yong Luo
- Guo-Wei Zuo
- Liang Chen
- Liang-Jun Yin
- Zhong-Liang Deng
- Bai-Cheng He
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Affiliations: Department of Orthopaedic Surgery, The Second Affiliated Hospital, Chongqing Medical University, Chongqing, P.R. China, Chongqing Key Laboratory of Biochemistry and Molecular Pharmacology, Chongqing Medical University, Chongqing, P.R. China, Key Laboratory of Diagnostic Medicine designated by the Chinese Ministry of Education, Chongqing Medical University, Chongqing, P.R. China
Published online on: May 22, 2014 https://doi.org/10.3892/ijo.2014.2456
Pages: 795-803

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Abstract

It has been reported that oridonin (ORI) can inhibit proliferation and induce apoptosis in various types of cancer cell lines. However, the exact mechanism for this function remains unclear. In this study, we investigated the proliferation inhibitory effect of ORI on human osteosarcoma (OS) 143B cells and dissected the possible molecular mechanism(s) underlying this effect. We demonstrated that ORI can inhibit proliferation, induce apoptosis and arrest the cell cycle in 143B cells. Using luciferase reporter assay, we found that the Wnt/β-catenin signaling was inhibited in 143B cells by ORI. Accordingly, the total protein levels and nuclear translocation of β-catenin were reduced by ORI treatment. ORI increased glycogen synthase kinase 3β (GSK3β) activity and upregulated Dickkopf-1 (Dkk-1) expression. We found that Dkk-1 overexpression or β-catenin knockdown can potentiate the proliferation inhibitory effect of ORI in 143B cells, while β-catenin overexpression attenuated this effect. Using the xenograft tumor model of human OS, we demonstrated that ORI effectively inhibited the growth of tumors. Histological examination showed that ORI inhibited cancer cell proliferation, decreased the expression of PNCA and β-catenin. Our findings suggest that ORI can inhibit 143B OS cell proliferation by downregulating Wnt/β-catenin signal transduction, which may be mediated by upregulating the Dkk-1 expression and/or enhancing the function of GSK3β. Therefore, ORI can be potentially used as an effective adjuvant agent for the clinical management of OS.

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1	Longhi A, Errani C, De Paolis M, Mercuri M and Bacci G: Primary bone osteosarcoma in the pediatric age: state of the art. Cancer Treat Rev. 32:423–436. 2006. View Article : Google Scholar : PubMed/NCBI
2	Ando K, Heymann MF, Stresing V, Mori K, Redini F and Heymann D: Current therapeutic strategies and novel approaches in osteosarcoma. Cancers. 5:591–616. 2013. View Article : Google Scholar : PubMed/NCBI
3	Abelson PH: Medicine from plants. Science. 247:5131990. View Article : Google Scholar
4	Liu YQ, Mu ZQ, You S, Tashiro S, Onodera S and Ikejima T: Fas/FasL signaling allows extracelluar-signal regulated kinase to regulate cytochrome c release in oridonin-induced apoptotic U937 cells. Biol Pharm Bull. 29:1873–1879. 2006. View Article : Google Scholar : PubMed/NCBI
5	Zhu Y, Xie L, Chen G, Wang H and Zhang R: Effects of oridonin on proliferation of HT29 human colon carcinoma cell lines both in vitro and in vivo in mice. Pharmazie. 62:439–444. 2007.PubMed/NCBI
6	Hsieh TC, Wijeratne EK, Liang JY, Gunatilaka AL and Wu JM: Differential control of growth, cell cycle progression, and expression of NF-kappaB in human breast cancer cells MCF-7, MCF-10A, and MDA-MB-231 by ponicidin and oridonin, diterpenoids from the chinese herb Rabdosia rubescens. Biochem Biophys Res Commun. 337:224–231. 2005. View Article : Google Scholar : PubMed/NCBI
7	Zhou GB, Kang H, Wang L, et al: Oridonin, a diterpenoid extracted from medicinal herbs, targets AML1-ETO fusion protein and shows potent antitumor activity with low adverse effects on t (8;21) leukemia in vitro and in vivo. Blood. 109:3441–3450. 2007. View Article : Google Scholar
8	Jin S, Shen JN, Wang J, Huang G and Zhou JG: Oridonin induced apoptosis through Akt and MAPKs signaling pathways in human osteosarcoma cells. Cancer Biol Ther. 6:261–268. 2007. View Article : Google Scholar : PubMed/NCBI
9	Behrens J and Lustig B: The Wnt connection to tumorigenesis. Int J Dev Biol. 48:477–487. 2004. View Article : Google Scholar : PubMed/NCBI
10	Barker N and Clevers H: Mining the Wnt pathway for cancer therapeutics. Nat Rev Drug Discov. 5:997–1014. 2006. View Article : Google Scholar : PubMed/NCBI
11	Lin YC, You L, Xu Z, et al: Wnt signaling activation and WIF-1 silencing in nasopharyngeal cancer cell lines. Biochem Biophys Res Commun. 341:635–640. 2006. View Article : Google Scholar : PubMed/NCBI
12	Weeraratna AT, Jiang Y, Hostetter G, et al: Wnt5a signaling directly affects cell motility and invasion of metastatic melanoma. Cancer Cell. 1:279–288. 2002. View Article : Google Scholar : PubMed/NCBI
13	Wissmann C, Wild PJ, Kaiser S, et al: WIF1, a component of the Wnt pathway, is down-regulated in prostate, breast, lung, and bladder cancer. J Pathol. 201:204–212. 2003. View Article : Google Scholar : PubMed/NCBI
14	Hoang BH, Kubo T, Healey JH, et al: Expression of LDL receptor-related protein 5 (LRP5) as a novel marker for disease progression in high-grade osteosarcoma. Int J Cancer. 109:106–111. 2004. View Article : Google Scholar : PubMed/NCBI
15	Iozzo RV, Eichstetter I and Danielson KG: Aberrant expression of the growth factor Wnt-5A in human malignancy. Cancer Res. 55:3495–3499. 1995.PubMed/NCBI
16	Lu BJ, Wang YQ, Wei XJ, et al: Expression of WNT-5a and ROR2 correlates with disease severity in osteosarcoma. Mol Med Rep. 5:1033–1036. 2012.PubMed/NCBI
17	Ma Y, Ren Y, Han EQ, et al: Inhibition of the Wnt-beta-catenin and Notch signaling pathways sensitizes osteosarcoma cells to chemotherapy. Biochem Biophys Res Commun. 431:274–279. 2013. View Article : Google Scholar : PubMed/NCBI
18	Kansara M, Tsang M, Kodjabachian L, et al: Wnt inhibitory factor 1 is epigenetically silenced in human osteosarcoma, and targeted disruption accelerates osteosarcomagenesis in mice. J Clin Invest. 119:837–851. 2009. View Article : Google Scholar : PubMed/NCBI
19	Gao FH, Hu XH, Li W, et al: Oridonin induces apoptosis and senescence in colorectal cancer cells by increasing histone hyperacetylation and regulation of p16, p21, p27 and c-myc. BMC Cancer. 10:6102010. View Article : Google Scholar : PubMed/NCBI
20	He BC, Chen L, Zuo GW, et al: Synergistic antitumor effect of the activated PPARgamma and retinoid receptors on human osteosarcoma. Clin Cancer Res. 16:2235–2245. 2010. View Article : Google Scholar : PubMed/NCBI
21	Wu K, Yang Q, Mu Y, et al: Berberine inhibits the proliferation of colon cancer cells by inactivating Wnt/beta-catenin signaling. Int J Oncol. 41:292–298. 2012.PubMed/NCBI
22	He TC, Zhou S, da Costa LT, Yu J, Kinzler KW and Vogelstein B: A simplified system for generating recombinant adenoviruses. Proc Natl Acad Sci USA. 95:2509–2514. 1998. View Article : Google Scholar : PubMed/NCBI
23	Tang N, Song WX, Luo J, et al: BMP-9-induced osteogenic differentiation of mesenchymal progenitors requires functional canonical Wnt/beta-catenin signalling. J Cell Mol Med. 13:2448–2464. 2009. View Article : Google Scholar : PubMed/NCBI
24	He BC, Gao JL, Luo X, et al: Ginsenoside Rg3 inhibits colorectal tumor growth through the down-regulation of Wnt/β-catenin signaling. Int J Oncol. 38:437–445. 2011.PubMed/NCBI
25	Park HR and Park YK: Expression of p53 protein, PCNA, and Ki-67 in osteosarcomas of bone. J Korean Med Sci. 10:360–367. 1995. View Article : Google Scholar : PubMed/NCBI
26	Davidson G and Niehrs C: Emerging links between CDK cell cycle regulators and Wnt signaling. Trends Cell Biol. 20:453–460. 2010. View Article : Google Scholar : PubMed/NCBI
27	Lustig B and Behrens J: The Wnt signaling pathway and its role in tumor development. J Cancer Res Clin Oncol. 129:199–221. 2003.PubMed/NCBI
28	Nakamura T, Hamada F, Ishidate T, et al: Axin, an inhibitor of the Wnt signalling pathway, interacts with beta-catenin, GSK-3beta and APC and reduces the beta-catenin level. Genes Cells. 3:395–403. 1998. View Article : Google Scholar : PubMed/NCBI
29	Cross DA, Alessi DR, Cohen P, Andjelkovich M and Hemmings BA: Inhibition of glycogen synthase kinase-3 by insulin mediated by protein kinase B. Nature. 378:785–789. 1995. View Article : Google Scholar
30	Bikkavilli RK, Feigin ME and Malbon CC: p38 mitogen-activated protein kinase regulates canonical Wnt-beta-catenin signaling by inactivation of GSK3beta. J Cell Sci. 121:3598–3607. 2008. View Article : Google Scholar
31	Li CY, Wang EQ, Cheng Y and Bao JK: Oridonin: an active diterpenoid targeting cell cycle arrest, apoptotic and autophagic pathways for cancer therapeutics. Int J Biochem Cell Biol. 43:701–704. 2011. View Article : Google Scholar : PubMed/NCBI
32	Zhang CL, Wu LJ, Zuo HJ, Tashiro S, Onodera S and Ikejima T: Cytochrome c release from oridonin-treated apoptotic A375-S2 cells is dependent on p53 and extracellular signal-regulated kinase activation. J Pharmacol Sci. 96:155–163. 2004. View Article : Google Scholar : PubMed/NCBI
33	Cheng Y, Qiu F, Ye YC, Tashiro S, Onodera S and Ikejima T: Oridonin induces G2/M arrest and apoptosis via activating ERK-p53 apoptotic pathway and inhibiting PTK-Ras-Raf-JNK survival pathway in murine fibrosarcoma L929 cells. Arch Biochem Biophys. 490:70–75. 2009. View Article : Google Scholar : PubMed/NCBI
34	Hu HZ, Yang YB, Xu XD, et al: Oridonin induces apoptosis via PI3K/Akt pathway in cervical carcinoma HeLa cell line. Acta Pharmacol Sin. 28:1819–1826. 2007. View Article : Google Scholar : PubMed/NCBI
35	Li X, Li X, Wang J, Ye Z and Li JC: Oridonin up-regulates expression of P21 and induces autophagy and apoptosis in human prostate cancer cells. Int J Biol Sci. 8:901–912. 2012. View Article : Google Scholar : PubMed/NCBI
36	Polakis P: Wnt signaling and cancer. Genes Dev. 14:1837–1851. 2000.
37	Baron R and Kneissel M: WNT signaling in bone homeostasis and disease: from human mutations to treatments. Nat Med. 19:179–192. 2013. View Article : Google Scholar : PubMed/NCBI
38	Reya T and Clevers H: Wnt signalling in stem cells and cancer. Nature. 434:843–850. 2005. View Article : Google Scholar : PubMed/NCBI
39	Haydon RC, Deyrup A, Ishikawa A, et al: Cytoplasmic and/or nuclear accumulation of the beta-catenin protein is a frequent event in human osteosarcoma. Int J Cancer. 102:338–342. 2002. View Article : Google Scholar : PubMed/NCBI
40	Rubin EM, Guo Y, Tu K, Xie J, Zi X and Hoang BH: Wnt inhibitory factor 1 decreases tumorigenesis and metastasis in osteosarcoma. Mol Cancer Ther. 9:731–741. 2010. View Article : Google Scholar : PubMed/NCBI
41	Guo Y, Rubin EM, Xie J, Zi X and Hoang BH: Dominant negative LRP5 decreases tumorigenicity and metastasis of osteosarcoma in an animal model. Clin Orthop Relat Res. 466:2039–2045. 2008. View Article : Google Scholar : PubMed/NCBI
42	Mishra R: Glycogen synthase kinase 3 beta: can it be a target for oral cancer. Mol Cancer. 9:1442010. View Article : Google Scholar : PubMed/NCBI
43	Wu J, Liao Q, He H, Zhong D and Yin K: TWIST interacts with beta-catenin signaling on osteosarcoma cell survival against cisplatin. Mol Carcinog. Dec 31–2012.(Epub ahead of print). View Article : Google Scholar
44	Xia JJ, Pei LB, Zhuang JP, et al: Celecoxib inhibits beta-catenin-dependent survival of the human osteosarcoma MG-63 cell line. J Int Med Res. 38:1294–1304. 2010. View Article : Google Scholar : PubMed/NCBI
45	Cai Y, Mohseny AB, Karperien M, Hogendoorn PC, Zhou G and Cleton-Jansen AM: Inactive Wnt/beta-catenin pathway in conventional high-grade osteosarcoma. J Pathol. 220:24–33. 2010. View Article : Google Scholar : PubMed/NCBI
46	Tang QL, Xie XB, Wang J, et al: Glycogen synthase kinase-3beta, NF-kappaB signaling, and tumorigenesis of human osteosarcoma. J Natl Cancer Inst. 104:749–763. 2012. View Article : Google Scholar : PubMed/NCBI
47	Zorn AM: Wnt signalling: antagonistic Dickkopfs. Curr Biol. 11:R592–R595. 2001. View Article : Google Scholar : PubMed/NCBI
48	Kawano Y and Kypta R: Secreted antagonists of the Wnt signalling pathway. J Cell Sci. 116:2627–2634. 2003. View Article : Google Scholar
49	Lin CH, Guo Y, Ghaffar S, et al: Dkk-3, a secreted wnt antagonist, suppresses tumorigenic potential and pulmonary metastasis in osteosarcoma. Sarcoma. 2013:1475412013.PubMed/NCBI
50	Hoang BH, Kubo T, Healey JH, et al: Dickkopf 3 inhibits invasion and motility of Saos-2 osteosarcoma cells by modulating the Wnt-beta-catenin pathway. Cancer Res. 64:2734–2739. 2004. View Article : Google Scholar : PubMed/NCBI