Norcantharidin blocks Wnt/β-catenin signaling via promoter demethylation of WIF-1 in glioma

Xie,Dajiang; Xie,Jixi; Wan,Yingfeng; Ma,Li; Qi,Xuchen; Wang,Kun; Yang,Shuxu

doi:10.3892/or.2016.4559

Norcantharidin blocks Wnt/β-catenin signaling via promoter demethylation of WIF-1 in glioma

Authors:
- Dajiang Xie
- Jixi Xie
- Yingfeng Wan
- Li Ma
- Xuchen Qi
- Kun Wang
- Shuxu Yang
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Affiliations: Department of Neurosurgery, Sir Run Run Shaw Hospital, College of Medical Sciences, Zhejiang University, Hangzhou, Zhejiang 310016, P.R. China
Published online on: January 14, 2016 https://doi.org/10.3892/or.2016.4559
Pages: 2191-2197

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Abstract

Glioma is one of the most common primary intracranial tumors, and the prognosis is poor even though much treatment management is employed. Wnt/β-catenin signaling has been reported to be associated with glioma. Norcantharidin (NCTD) is the demethylated analog of cantharidin isolated from blister beetles, and it is reported to possess anticancer activity but less nephrotoxicity than cantharidin. Accordingly, we aimed to investigate NCTD as an anti-neoplastic drug that inhibits the Wnt/β‑catenin pathway via promoter demethylation of Wnt inhibitory factor-1 (WIF-1) in glioma growth in vitro. In the present study, we report that NCTD inhibited cell proliferation, induced apoptosis and cell cycle arrest, and suppressed cell migration and invasion in vitro. Moreover, we observed that the expression levels of WIF-1 mRNA and protein in the NCTD-treated cells were increased significantly compared with these levels in the negative control (NC) cells. Promoter demethylation was observed in the NCTD‑treated cells. In contrast, aberrant methylation was observed in the NC cells. Additionally, more investigation revealed that NCTD suppressed activity of Wnt/β-catenin signaling and transcription of β-catenin/TCF-4. Furthermore, the expression of apoptosis-related proteins Bcl-2 and cleaved caspase-3 indicated significant cell apoptosis. We provide initial evidence that NCTD reactivates WIF-1 from a methylation state, and downregulates canonical Wnt/β-catenin signaling. Our findings revealed that NCTD is effective for glioma in vitro and may be a new therapeutic option in vivo.

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1	Chen W, Wu Q, Mo L and Nassi M: Intra-arterial chemotherapy is not superior to intravenous chemotherapy for malignant gliomas: A systematic review and meta-analysis. Eur Neurol. 70:124–132. 2013. View Article : Google Scholar : PubMed/NCBI
2	Bregy A, Shah AH, Diaz MV, Pierce HE, Ames PL, Diaz D and Komotar RJ: The role of Gliadel wafers in the treatment of high-grade gliomas. Expert Rev Anticancer Ther. 13:1453–1461. 2013. View Article : Google Scholar : PubMed/NCBI
3	Yin AA, Cai S, Dong Y, Zhang LH, Liu BL, Cheng JX and Zhang X: A meta-analysis of temozolomide versus radiotherapy in elderly glioblastoma patients. J Neurooncol. 116:315–324. 2014. View Article : Google Scholar
4	Yin AA, Zhang LH, Cheng JX, Dong Y, Liu BL, Han N and Zhang X: Radiotherapy plus concurrent or sequential temozolomide for glioblastoma in the elderly: A meta-analysis. PLoS One. 8:e742422013. View Article : Google Scholar : PubMed/NCBI
5	Wang Y and Jiang T: Understanding high grade glioma: Molecular mechanism, therapy and comprehensive management. Cancer Lett. 331:139–146. 2013. View Article : Google Scholar : PubMed/NCBI
6	Schneider AJ, Branam AM and Peterson RE: Intersection of AHR and Wnt signaling in development, health, and disease. Int J Mol Sci. 15:17852–17885. 2014. View Article : Google Scholar : PubMed/NCBI
7	Kim SA, Kwak J, Nam HY, Chun SM, Lee BW, Lee HJ, Khang SK and Kim SW: Promoter methylation of WNT inhibitory factor-1 and expression pattern of WNT/β-catenin pathway in human astrocytoma: Pathologic and prognostic correlations. Mod Pathol. 26:626–639. 2013. View Article : Google Scholar : PubMed/NCBI
8	Wu J, Fang J, Yang Z, Chen F, Liu J and Wang Y: Wnt inhibitory factor-1 regulates glioblastoma cell cycle and proliferation. J Clin Neurosci. 19:1428–1432. 2012. View Article : Google Scholar : PubMed/NCBI
9	Yang Z and Wang Y, Fang J, Chen F, Liu J, Wu J and Wang Y: Expression and aberrant promoter methylation of Wnt inhibitory factor-1 in human astrocytomas. J Exp Clin Cancer Res. 29:262010. View Article : Google Scholar : PubMed/NCBI
10	Jiang YM, Meng ZZ, Yue GX and Chen JX: Norcantharidin induces HL-60 cell apoptosis in vitro. Evid Based Complement Alternat Med. 2012:1542712012. View Article : Google Scholar
11	Clevers H, Loh KM and Nusse R: Stem cell signaling. An integral program for tissue renewal and regeneration: Wnt signaling and stem cell control. Science. 346:12480122014. View Article : Google Scholar : PubMed/NCBI
12	Jamieson C, Sharma M and Henderson BR: Targeting the β-catenin nuclear transport pathway in cancer. Semin Cancer Biol. 27:20–29. 2014. View Article : Google Scholar : PubMed/NCBI
13	Le PN, McDermott JD and Jimeno A: Targeting the Wnt pathway in human cancers: therapeutic targeting with a focus on OMP-54F28. Pharmacol Ther. 146:1–11. 2015. View Article : Google Scholar
14	Bilir B, Kucuk O and Moreno CS: Wnt signaling blockage inhibits cell proliferation and migration, and induces apoptosis in triple-negative breast cancer cells. J Transl Med. 11:2802013. View Article : Google Scholar : PubMed/NCBI
15	Chen X, Hu W, Xie B, Gao H, Xu C and Chen J: Downregulation of SCAI enhances glioma cell invasion and stem cell like phenotype by activating Wnt/β-catenin signaling. Biochem Biophys Res Commun. 448:206–211. 2014. View Article : Google Scholar : PubMed/NCBI
16	Schüle R, Dictus C, Campos B, Wan F, Felsberg J, Ahmadi R, Centner FS, Grabe N, Reifenberger G, Bermejo JL, et al: Potential canonical wnt pathway activation in high-grade astrocytomas. Sci World J. 2012:6973132012. View Article : Google Scholar
17	Wang K, Wang X, Zou J, Zhang A, Wan Y, Pu P, Song Z, Qian C, Chen Y, Yang S, et al: miR-92b controls glioma proliferation and invasion through regulating Wnt/beta-catenin signaling via Nemo-like kinase. Neuro-oncol. 15:578–588. 2013. View Article : Google Scholar : PubMed/NCBI
18	Delic S, Lottmann N, Stelzl A, Liesenberg F, Wolter M, Götze S, Zapatka M, Shiio Y, Sabel MC, Felsberg J, et al: MiR-328 promotes glioma cell invasion via SFRP1-dependent Wnt-signaling activation. Neuro-oncol. 16:179–190. 2014. View Article : Google Scholar :
19	Liu D, Shi P, Yin X, Chen Z and Zhang X: Effect of norcantharidin on the human breast cancer Bcap-37 cells. Connect Tissue Res. 53:508–512. 2012. View Article : Google Scholar : PubMed/NCBI
20	Lee YC, Lee LM, Yang CH, Lin AM, Huang YC, Hsu CC, Chen MS, Chi CW, Yin PH, Kuo CD, et al: Norcantharidin suppresses cell growth and migration with enhanced anticancer activity of gefitinib and cisplatin in human non-small cell lung cancer cells. Oncol Rep. 29:237–243. 2013.
21	Liao HF, Chen YJ, Chou CH, Wang FW and Kuo CD: Norcantharidin induces cell cycle arrest and inhibits progression of human leukemic Jurkat T cells through mitogen-activated protein kinase-mediated regulation of interleukin-2 production. Toxicol In Vitro. 25:206–212. 2011. View Article : Google Scholar
22	Liu S, Yu H, Kumar SM, Martin JS, Bing Z, Sheng W, Bosenberg M and Xu X: Norcantharidin induces melanoma cell apoptosis through activation of TR3 dependent pathway. Cancer Biol Ther. 12:1005–1014. 2011. View Article : Google Scholar : PubMed/NCBI
23	Shen B, He PJ and Shao CL: Norcantharidin induced DU145 cell apoptosis through ROS-mediated mitochondrial dysfunction and energy depletion. PLoS One. 8:e846102013. View Article : Google Scholar : PubMed/NCBI
24	Shou LM, Zhang QY, Li W, Xie X, Chen K, Lian L, Li ZY, Gong FR, Dai KS, Mao YX, et al: Cantharidin and norcantharidin inhibit the ability of MCF-7 cells to adhere to platelets via protein kinase C pathway-dependent downregulation of α2 integrin. Oncol Rep. 30:1059–1066. 2013.PubMed/NCBI
25	An WW, Gong XF, Wang MW, Tashiro S, Onodera S and Ikejima T: Norcantharidin induces apoptosis in HeLa cells through caspase, MAPK, and mitochondrial pathways. Acta Pharmacol Sin. 25:1502–1508. 2004.PubMed/NCBI
26	Yu CC, Ko FY, Yu CS, Lin CC, Huang YP, Yang JS, Lin JP and Chung JG: Norcantharidin triggers cell death and DNA damage through S-phase arrest and ROS-modulated apoptotic pathways in TSGH 8301 human urinary bladder carcinoma cells. Int J Oncol. 41:1050–1060. 2012.PubMed/NCBI
27	Zhang L, Ji Q, Liu X, Chen X, Chen Z, Qiu Y, Sun J, Cai J, Zhu H and Li Q: Norcantharidin inhibits tumor angiogenesis via blocking VEGFR2/MEK/ERK signaling pathways. Cancer Sci. 104:604–610. 2013. View Article : Google Scholar : PubMed/NCBI
28	Zhang S, Li G, Ma X, Wang Y, Liu G, Feng L, Zhao Y, Zhang G, Wu Y, Ye X, et al: Norcantharidin enhances ABT-737-induced apoptosis in hepatocellular carcinoma cells by transcriptional repression of Mcl-1. Cell Signal. 24:1803–1809. 2012. View Article : Google Scholar : PubMed/NCBI
29	Zhang JT, Sun W, Zhang WZ, Ge CY, Liu ZY, Zhao ZM, Lu XS and Fan YZ: Norcantharidin inhibits tumor growth and vasculogenic mimicry of human gallbladder carcinomas by suppression of the PI3-K/MMPs/Ln-5γ2 signaling pathway. BMC Cancer. 14:1932014. View Article : Google Scholar
30	Chen S, Wan P, Ding W, Li F, He C, Chen P, Li H, Hu Z, Tan W and Li J: Norcantharidin inhibits DNA replication and induces mitotic catastrophe by degrading initiation protein Cdc6. Int J Mol Med. 32:43–50. 2013.PubMed/NCBI
31	Pitre A, Davis N, Paul M, Orr AW and Skalli O: Synemin promotes AKT-dependent glioblastoma cell proliferation by antagonizing PP2A. Mol Biol Cell. 23:1243–1253. 2012. View Article : Google Scholar : PubMed/NCBI
32	Zheng J, Du W, Song LJ, Zhang R, Sun LG, Chen FG and Wei XT: Norcantharidin induces growth inhibition and apoptosis of glioma cells by blocking the Raf/MEK/ERK pathway. World J Surg Oncol. 12:2072014. View Article : Google Scholar : PubMed/NCBI
33	Wu MY, Xie X, Xu ZK, Xie L, Chen Z, Shou LM, Gong FR, Xie YF, Li W and Tao M: PP2A inhibitors suppress migration and growth of PANC-1 pancreatic cancer cells through inhibition on the Wnt/β-catenin pathway by phosphorylation and degradation of β-catenin. Oncol Rep. 32:513–522. 2014.PubMed/NCBI
34	Chuang KA, Lieu CH, Tsai WJ, Wu MH, Chen YC, Liao JF, Wang CC and Kuo YC: Evaluation of anti-Wnt/β-catenin signaling agents by pGL4-TOP transfected stable cells with a luciferase reporter system. Braz J Med Biol Res. 43:931–941. 2010. View Article : Google Scholar : PubMed/NCBI
35	Hsieh CH, Chao KS, Liao HF and Chen YJ: Norcantharidin, derivative of cantharidin, for cancer stem cells. Evid Based Complement Alternat Med. 2013:8386512013. View Article : Google Scholar : PubMed/NCBI
36	Liu YL, Yang HP, Zhou XD, Gong L, Tang CL and Wang HJ: The hypomethylation agent bisdemethoxycurcumin acts on the WIF-1 promoter, inhibits the canonical Wnt pathway and induces apoptosis in human non-small-cell lung cancer. Curr Cancer Drug Targets. 11:1098–1110. 2011. View Article : Google Scholar : PubMed/NCBI