Rescued expression of WIF-1 in gallbladder cancer inhibits tumor growth and induces tumor cell apoptosis with altered expression of proteins

Huang,Yan; Du,Qiang; Wu,Weibao; She,Feifei; Chen,Yanling

doi:10.3892/mmr.2016.5532

Rescued expression of WIF-1 in gallbladder cancer inhibits tumor growth and induces tumor cell apoptosis with altered expression of proteins

Authors:
- Yan Huang
- Qiang Du
- Weibao Wu
- Feifei She
- Yanling Chen
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Affiliations: Department of Hepatobiliary Surgery, The Affiliated Union Hospital of Fujian Medical University, Fuzhou, Fujian 350001, P.R. China, Key Laboratory of Ministry of Education for Gastrointestinal Cancer, School of Basic Medical Sciences, Fujian Medical University, Fuzhou, Fujian 350001, P.R. China
Published online on: July 19, 2016 https://doi.org/10.3892/mmr.2016.5532
Pages: 2573-2581
Copyright: © Huang et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

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Abstract

As a highly conserved metabolic pathway, the Wnt signaling pathway is involved in cell differentiation, proliferation and several other processes. In normal cells, this pathway is suppressed, and abnormal activation is often associated with tumor occurrence and development. In certain types of tumor, Wnt inhibitory factor 1 (WIF‑1), an inhibitor of the Wnt pathway, inhibits tumor growth. However, the effect of the expression of WIF-1 on gallbladder cancer remains to be fully elucidated. In the current study, reverse transcription‑quantitative polymerase chain reaction and western blotting were conducted. The present study demonstrated that, in gallbladder cancer, WIF‑1 generally exhibited low levels of expression as a result of gene promoter methylation. Treatment with the drug, 5-aza-2-deoxycytidine, increased the expression of WIF‑1 in the GBC‑SD gallbladder cell line. In addition, a WIF‑1‑expression plasmid was transfected into GBC‑SD cells, and it was found that cell proliferation, invasion and metastasis declined significantly, whereas the apoptotic rate increased. A nude mouse tumor transplantation experiment showed that the oncogenicity of the GBC‑SD cells expressing WIF‑1 was substantially lower, compared with that of the untransfected GBC‑SD cells and of GBD‑SD cells expressing the control plasmid. A fluorescent protein chip experiment showed that the restored expression of WIF‑1 affected the expression of several cellular proteins. These alterations may explain the different biological behavior of the tumor cells expressing WIF‑1. As an effective inhibitory factor of the Wnt signaling pathway, WIF‑1 modulated the expression of proteins controlling the proliferation, apoptosis and metastasis of gallbladder tumor cells, thus suppressing the tumor. Therefore, WIF‑1 may be an effective treatment target for gallbladder cancer.

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1	Donohue JH, Stewart AK and Menck HR: The national cancer data base report on carcinoma of the gallbladder, 1989–1995. Cancer. 83:2618–2628. 1998. View Article : Google Scholar
2	Coburn NG, Cleary SP, Tan JC and Law CH: Surgery for gallbladder cancer: A population-based analysis. J Am Coll Surg. 207:371–382. 2008. View Article : Google Scholar : PubMed/NCBI
3	Zhu AX, Hong TS, Hezel AF and Kooby DA: Current management of gallbladder carcinoma. Oncologist. 15:168–181. 2010. View Article : Google Scholar : PubMed/NCBI
4	Nusse R and Varmus HE: Many tumors induced by the mouse mammary tumor virus contain a provirus integrated in the same region of the host genome. Cell. 31:99–109. 1982. View Article : Google Scholar : PubMed/NCBI
5	Wodarz A and Nusse R: Mechanisms of Wnt signaling in development. Annu Rev Cell Dev Biol. 14:59–88. 1998. View Article : Google Scholar
6	Clevers H: Wnt/beta-catenin signaling in development and disease. Cell. 127:469–480. 2006. View Article : Google Scholar : PubMed/NCBI
7	Taketo MM: Wnt signaling and gastrointestinal tumorigenesis in mouse models. Oncogene. 25:7522–7530. 2006. View Article : Google Scholar : PubMed/NCBI
8	Van Scoyk M, Randall J, Sergew A, Williams LM, Tennis M and Winn RA: Wnt signaling pathway and lung disease. Transl Res. 151:175–180. 2008. View Article : Google Scholar : PubMed/NCBI
9	Karamboulas C and Ailles L: Developmental signaling pathways in cancer stem cells of solid tumors. Biochim Biophys Acta. 1830:2481–2495. 2013. View Article : Google Scholar
10	Hsieh JC, Kodjabachian L, Rebbert ML, Rattner A, Smallwood PM, Samos CH, Nusse R, Dawid IB and Nathans J: A new secreted protein that binds to Wnt proteins and inhibits their activities. Nature. 398:431–436. 1999. View Article : Google Scholar
11	Liepinsh E, Bányai L, Patthy L and Otting G: NMR structure of the WIF domain of the human Wnt-inhibitory factor-1. J Mol Biol. 357:942–950. 2006. View Article : Google Scholar : PubMed/NCBI
12	Kawano Y and Kypta R: Secreted antagonists of the Wnt signaling pathway. J Cell Sci. 116:2627–2634. 2003. View Article : Google Scholar : PubMed/NCBI
13	Kim AS, Lowenstein DH and Pleasure SJ: Wnt receptors and Wnt inhibitors are expressed in gradients in the developing telencephalon. Mech Dev. 103:167–172. 2001. View Article : Google Scholar : PubMed/NCBI
14	Wissmann C, Wild PJ, Kaiser S, Roepcke S, Stoehr R, Woenckhaus M, Kristiansen G, Hsieh JC, Hofstaedter F, Hartmann A, et al: WIF1, a component of the Wnt pathway, is downregulated in prostate, breast, lung and bladder cancer. J Pathol. 201:204–212. 2003. View Article : Google Scholar : PubMed/NCBI
15	Licchesi JD, Westra WH, Hooker CM, Machida EO, Baylin SB and Herman JG: Epigenetic alteration of Wnt pathway antagonists in progressive glandular neoplasia of the lung. Carcinogenesis. 29:895–904. 2008. View Article : Google Scholar : PubMed/NCBI
16	Byun T, Karimi M, Marsh JL, Milovanovic T, Lin F and Holcombe RF: Expression of secreted Wnt antagonists in gastrointestinal tissues: Potential role in stem cell homeostasis. J Clin Pathol. 58:515–519. 2005. View Article : Google Scholar : PubMed/NCBI
17	Frommer M, McDonald LE, Millar DS, Collis CM, Watt F, Grigg GW, Molloy PL and Paul CL: A genomic sequencing protocol that yields a positive display of 5-methylcytosine residues in individual DNA strands. Proc Natl Acad Sci USA. 89:1827–1831. 1992. View Article : Google Scholar : PubMed/NCBI
18	Behrens J and Lustig B: The Wnt connection to tumorigenesis. Int J Dev Biol. 48:477–487. 2004. View Article : Google Scholar : PubMed/NCBI
19	Taniguchi H, Yamamoto H, Hirata T, Miyamoto N, Oki M, Nosho K, Adachi Y, Endo T, Imai K and Shinomura Y: Frequent epigenetic inactivation of Wnt inhibitory factor-1 in human gastrointestinal cancers. Oncogene. 24:7946–7952. 2005. View Article : Google Scholar : PubMed/NCBI
20	Schroeder M and Mass MJ: CpG methylation inactivates the transcriptional activity of the promoter of the human p53 tumor suppressor gene. Biochem Biophys Res Commun. 235:403–406. 1997. View Article : Google Scholar : PubMed/NCBI
21	Holmes R and Soloway PD: Regulation of imprinted DNA methylation. Cytogenet Genome Res. 113:122–129. 2006. View Article : Google Scholar : PubMed/NCBI
22	Das PM and Singal R: DNA methylation and cancer. J Clin Oncol. 22:4632–4642. 2004. View Article : Google Scholar : PubMed/NCBI
23	Batra S, Shi Y, Kuchenbecker KM, He B, Reguart N, Mikami I, You L, Xu Z, Lin YC, Clément G and Jablons DM: Wnt inhibitory factor-1, a Wnt antagonist, is silenced by promoter hypermethylation in malignant pleural mesothelioma. Biochem Biophys Res Commun. 342:1228–1232. 2006. View Article : Google Scholar : PubMed/NCBI
24	Wang Y, Zhu CS, Bi KH, Xu WW, Dong L and Hou M: Study of WIF-1 promoter methylation with expressions of β-catenin in acute leukemia. Zhonghua Yi Xue Za Zhi. 91:2858–2860. 2011.In Chinese.
25	Alvarez C, Tapia T, Cornejo V, Fernandez W, Muñoz A, Camus M, Alvarez M, Devoto L and Carvallo P: Silencing of tumor suppressor genes RASSF1A, SLIT2 and WIF1 by promoter hypermethylation in hereditary breast cancer. Mol Carcinog. 52:475–487. 2013. View Article : Google Scholar
26	Chan SL, Cui Y, van Hasselt A, Li H, Srivastava G, Jin H, Ng KM, Wang Y, Lee KY, Tsao GS, et al: The tumor suppressor Wnt inhibitory factor 1 is frequently methylated in nasopharyngeal and esophageal carcinomas. Lab Invest. 87:644–650. 2007. View Article : Google Scholar : PubMed/NCBI
27	Palmisano WA, Divine KK, Saccomanno G, Gilliland FD, Baylin SB, Herman JG and Belinsky SA: Predicting lung cancer by detecting aberrant promoter methylation in sputum. Cancer Res. 60:5954–5958. 2000.PubMed/NCBI
28	Belinsky SA, Nikula KJ, Palmisano WA, Michels R, Saccomanno G, Gabrielson E, Baylin SB and Herman JG: Aberrant methylation of p16 (INK4a) is an early event in lung cancer and a potential biomarker for early diagnosis. Proc Natl Acad Sci USA. 95:11891–11896. 1998. View Article : Google Scholar
29	Bryan J, Kantarjian H, Garcia-Manero G and Jabbour E: Pharmacokinetic evaluation of decitabine for the treatment of leukemia. Expert Opin Drug Metab Toxicol. 7:661–672. 2011. View Article : Google Scholar : PubMed/NCBI
30	Gollob JA, Sciambi CJ, Peterson BL, Richmond T, Thoreson M, Moran K, Dressman HK, Jelinek J and Issa JP: Phase I trial of sequential low-dose 5-aza-2′-deoxycytidine plus high-dose intravenous bolus interleukin-2 in patients with melanoma or renal cell carcinoma. Clin Cancer Res. 12:4619–4627. 2006. View Article : Google Scholar
31	Kawakami K, Hirata H, Yamamura S, Kikuno N, Saini S, Majid S, Tanaka Y, Kawamoto K, Enokida H, Nakagawa M and Dahiya R: Functional significance of Wnt inhibitory factor-1 gene in kidney cancer. Cancer Res. 69:8603–8610. 2009. View Article : Google Scholar : PubMed/NCBI
32	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
33	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
34	Kaushik SP: Current perspectives in gallbladder carcinoma. J Gastroenterol Hepatol. 16:848–854. 2001. View Article : Google Scholar
35	Benaud C, Dickson RB and Thompson EW: Roles of the matrix metalloproteinases in mammary gland development and cancer. Breast Cancer Res Treat. 50:97–116. 1998. View Article : Google Scholar : PubMed/NCBI
36	Kim HB, Kim CK, Iijima K, Kobayashi T and Kita H: Protein microarray analysis in patients with asthma: Elevation of the chemokine PARC/CCL18 in sputum. Chest. 135:295–302. 2009. View Article : Google Scholar
37	Wu DJ, Qian MJ, Rong RM, Xu M and Zhu TY: Expression of inflammation cytokines and network analysis in acute rejection of renal transplantation. Zhonghua Yi Xue Za Zhi. 92:2976–2979. 2012.In Chinese.
38	Enomoto-Iwamoto M, Kitagaki J, Koyama E, Tamamura Y, Wu C, Kanatani N, Koike T, Okada H, Komori T, Yoneda T, et al: The Wnt antagonist Frzb-1 regulates chondrocyte maturation and long bone development during limb skeletogenesis. Dev Biol. 251:142–156. 2002. View Article : Google Scholar : PubMed/NCBI
39	Dun Y, Yang Y, Xiong Z, Feng M, Zhang Y, Wang M, Xiang J, Li G and Ma R: Induction of Dickkopf-1 contributes to the neurotoxicity of MPP (+) in PC12 cells via inhibition of the canonical Wnt signaling pathway. Neuropharmacology. 67:168–175. 2013. View Article : Google Scholar
40	Chen B, Ma X, Liu S, Zhao W and Wu J: Inhibition of lung cancer cells growth, motility and induction of apoptosis by Klotho, a novel secreted Wnt antagonist, in a dose-dependent manner. Cancer Biol Ther. 13:1221–1228. 2012. View Article : Google Scholar : PubMed/NCBI