Ape1 regulates WNT/β-catenin signaling through its redox functional domain in pancreatic cancer cells

Jiang,Shaojie; Zhu,Lina; Tang,Haimei; Zhang,Miaofeng; Chen,Zhihua; Fei,Jian; Han,Baosan; Zou,Gang-Ming

doi:10.3892/ijo.2015.3048

Ape1 regulates WNT/β-catenin signaling through its redox functional domain in pancreatic cancer cells

Authors:
- Shaojie Jiang
- Lina Zhu
- Haimei Tang
- Miaofeng Zhang
- Zhihua Chen
- Jian Fei
- Baosan Han
- Gang-Ming Zou
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Affiliations: Department of Radiology, Sir Run Run Shaw Hospital, School of Medicine, Zhejiang University, Hangzhou, Zhejiang 310016, P.R. China, Department of Ophthalmology, Renji Hospital, School of Medicine, Shanghai Jiao Tong University, Shanghai 200127, P.R. China, Biomedical Research Center, Sir Run Run Shaw Hospital, School of Medicine, Zhejiang University, Hangzhou, Zhejiang 310016, P.R. China, Department of Orthopaedics, Second Affiliated Hospital, School of Medicine, Zhejiang University, Hangzhou, Zhejiang 310009, P.R. China, Xin Hua Hospital, Shanghai Key Laboratory for Pediatrics Gastroenterology and Nutrition, Shanghai Institute for Pediatrics Research, Shanghai Jiao Tong University School of Medicine, Shanghai 200092, P.R. China, Department of Surgery, Ruijin Hospital, School of Medicine, Shanghai Jiao Tong University, Shanghai 200025, P.R. China, Department of Surgery, Xinhua Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai 200092, P.R. China
Published online on: June 12, 2015 https://doi.org/10.3892/ijo.2015.3048
Pages: 610-620

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Abstract

Apurinic/apyrimidinic endonuclease 1/redox factor-1 (Ape1/Ref-1, Ape1) is a multifunctional protein that is upregulated in human pancreatic cancer. Ape1 redox domain plays an essential role in regulating the effects of reactive oxygen species (ROS) generated during physiological metabolism and pathological stress. In the present study, we explored whether Ape1 and ROS affect WNT/β-catenin signaling. We used E3330, a small molecule inhibitor of the redox activity of Ape1, and a siRNA approach to knock down Ape1, in two human pancreatic cancer cell lines. Inhibition of Ape1 resulted in growth suppression of pancreatic cancer cells, increased ROS levels, upregulation of β-catenin and c-myc and downregulation of cyclin D1. Consistent with these data, overexpression of Ape1 in pancreatic cancer cells reduced ROS and c-myc levels and increased cyclin D1 levels. Moreover, treatment of pancreatic cancer cells with H2O2 to induce oxidative stress resulted in upregulated ROS levels, decreased Ape1 at both the mRNA and protein level, and alterations in WNT/β-catenin pathway components. Finally, treatment of pancreatic cancer cells with the WNT/β-catenin inhibitor IWR-1 resulted in growth inhibition, which was greatly enhanced when combined with E3330 treatment. In summary, our results demonstrate that ROS is an important intracellular messenger that can modulate WNT/β‑catenin signaling. The present study provides interesting new insight into crosstalk between the redox function of Ape1 and WNT/β-catenin signaling in cancer cells. Furthermore, our data show that the combination of Ape1 and WNT inhibitors enhanced the inhibition of pancreatic cell proliferation. These results provide a promising novel therapeutic strategy for treating pancreatic cancer in future.

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1	Haeno H, Gonen M, Davis MB, Herman JM, Iacobuzio-Donahue CA and Michor F: Computational modeling of pancreatic cancer reveals kinetics of metastasis suggesting optimum treatment strategies. Cell. 148:362–375. 2012. View Article : Google Scholar : PubMed/NCBI
2	Fishel ML, Jiang Y, Rajeshkumar NV, Scandura G, Sinn AL, He Y, Shen C, Jones DR, Pollok KE, Ivan M, et al: Impact of APE1/Ref-1 redox inhibition on pancreatic tumor growth. Mol Cancer Ther. 10:1698–1708. 2011. View Article : Google Scholar : PubMed/NCBI
3	Guillaumond F, Leca J, Olivares O, Lavaut MN, Vidal N, Berthezène P, Dusetti NJ, Loncle C, Calvo E, Turrini O, et al: Strengthened glycolysis under hypoxia supports tumor symbiosis and hexosamine biosynthesis in pancreatic adenocarcinoma. Proc Natl Acad Sci USA. 110:3919–3924. 2013. View Article : Google Scholar : PubMed/NCBI
4	Zou GM, Karikari C, Kabe Y, Handa H, Anders RA and Maitra A: The Ape-1/Ref-1 redox antagonist E3330 inhibits the growth of tumor endothelium and endothelial progenitor cells: Therapeutic implications in tumor angiogenesis. J Cell Physiol. 219:209–218. 2009. View Article : Google Scholar
5	Cannito S, Novo E, Compagnone A, Valfrè di Bonzo L, Busletta C, Zamara E, Paternostro C, Povero D, Bandino A, Bozzo F, et al: Redox mechanisms switch on hypoxia-dependent epithelial-mesenchymal transition in cancer cells. Carcinogenesis. 29:2267–2278. 2008. View Article : Google Scholar : PubMed/NCBI
6	Zhang Y, Morris JP IV, Yan W, Schofield HK, Gurney A, Simeone DM, Millar SE, Hoey T, Hebrok M and Pasca di Magliano M: Canonical wnt signaling is required for pancreatic carcinogenesis. Cancer Res. 73:4909–4922. 2013. View Article : Google Scholar : PubMed/NCBI
7	Korswagen HC: Regulation of the Wnt/beta-catenin pathway by redox signaling. Dev Cell. 10:687–688. 2006. View Article : Google Scholar : PubMed/NCBI
8	Zou GM, Luo MH, Reed A, Kelley MR and Yoder MC: Ape1 regulates hematopoietic differentiation of embryonic stem cells through its redox functional domain. Blood. 109:1917–1922. 2007. View Article : Google Scholar
9	Mol CD, Izumi T, Mitra S and Tainer JA: DNA-bound structures and mutants reveal abasic DNA binding by APE1 and DNA repair coordination (corrected). Nature. 403:451–456. 2000. View Article : Google Scholar : PubMed/NCBI
10	Luo M, Delaplane S, Jiang A, Reed A, He Y, Fishel M, Nyland RL II, Borch RF, Qiao X, Georgiadis MM, et al: Role of the multifunctional DNA repair and redox signaling protein Ape1/Ref-1 in cancer and endothelial cells: Small-molecule inhibition of the redox function of Ape1. Antioxid Redox Signal. 10:1853–1867. 2008. View Article : Google Scholar : PubMed/NCBI
11	Vasko MR, Guo C and Kelley MR: The multifunctional DNA repair/redox enzyme Ape1/Ref-1 promotes survival of neurons after oxidative stress. DNA Repair (Amst). 4:367–379. 2005. View Article : Google Scholar
12	Fishel ML, He Y, Reed AM, Chin-Sinex H, Hutchins GD, Mendonca MS and Kelley MR: Knockdown of the DNA repair and redox signaling protein Ape1/Ref-1 blocks ovarian cancer cell and tumor growth. DNA Repair (Amst). 7:177–186. 2008. View Article : Google Scholar
13	Tell G, Quadrifoglio F, Tiribelli C and Kelley MR: The many functions of APE1/Ref-1: Not only a DNA repair enzyme. Antioxid Redox Signal. 11:601–620. 2009. View Article : Google Scholar
14	Kelley MR and Fishel ML: DNA repair proteins as molecular targets for cancer therapeutics. Anticancer Agents Med Chem. 8:417–425. 2008. View Article : Google Scholar : PubMed/NCBI
15	Georgiadis MM, Luo M, Gaur RK, Delaplane S, Li X and Kelley MR: Evolution of the redox function in mammalian apurinic/apyrimidinic endonuclease. Mutat Res. 643:54–63. 2008. View Article : Google Scholar : PubMed/NCBI
16	Fishel ML, He Y, Smith ML and Kelley MR: Manipulation of base excision repair to sensitize ovarian cancer cells to alkylating agent temozolomide. Clin Cancer Res. 13:260–267. 2007. View Article : Google Scholar : PubMed/NCBI
17	Madlener S, Ströbel T, Vose S, Saydam O, Price BD, Demple B and Saydam N: Essential role for mammalian apurinic/apyrimidinic (AP) endonuclease Ape1/Ref-1 in telomere maintenance. Proc Natl Acad Sci USA. 110:17844–17849. 2013. View Article : Google Scholar : PubMed/NCBI
18	Jeon BH, Gupta G, Park YC, Qi B, Haile A, Khanday FA, Liu YX, Kim JM, Ozaki M, White AR, et al: Apurinic/apyrimidinic endonuclease 1 regulates endothelial NO production and vascular tone. Circ Res. 95:902–910. 2004. View Article : Google Scholar : PubMed/NCBI
19	Vascotto C, Fantini D, Romanello M, Cesaratto L, Deganuto M, Leonardi A, Radicella JP, Kelley MR, D’Ambrosio C, Scaloni A, et al: APE1/Ref-1 interacts with NPM1 within nucleoli and plays a role in the rRNA quality control process. Mol Cell Biol. 29:1834–1854. 2009. View Article : Google Scholar : PubMed/NCBI
20	Poletto M, Vascotto C, Scognamiglio PL, Lirussi L, Marasco D and Tell G: Role of the unstructured N-terminal domain of the hAPE1 (human apurinic/apyrimidinic endonuclease 1) in the modulation of its interaction with nucleic acids and NPM1 (nucleophosmin). Biochem J. 452:545–557. 2013. View Article : Google Scholar : PubMed/NCBI
21	Vascotto C, Lirussi L, Poletto M, Tiribelli M, Damiani D, Fabbro D, Damante G, Demple B, Colombo E and Tell G: Functional regulation of the apurinic/apyrimidinic endonuclease 1 by nucleophosmin: Impact on tumor biology. Oncogene. 33:2876–2887. 2014. View Article : Google Scholar
22	Huang E, Qu D, Zhang Y, Venderova K, Haque ME, Rousseaux MW, Slack RS, Woulfe JM and Park DS: The role of Cdk5-mediated apurinic/apyrimidinic endonuclease 1 phosphorylation in neuronal death. Nat Cell Biol. 12:563–571. 2010. View Article : Google Scholar : PubMed/NCBI
23	Zou GM and Maitra A: Small-molecule inhibitor of the AP endonuclease 1/REF-1 E3330 inhibits pancreatic cancer cell growth and migration. Mol Cancer Ther. 7:2012–2021. 2008. View Article : Google Scholar : PubMed/NCBI
24	Wang YT, Tzeng DW, Wang CY, Hong JY and Yang JL: APE1/Ref-1 prevents oxidative inactivation of ERK for G1-to-S progression following lead acetate exposure. Toxicology. 305:120–129. 2013. View Article : Google Scholar : PubMed/NCBI
25	Lee HM, Yuk JM, Shin DM, Yang CS, Kim KK, Choi DK, Liang ZL, Kim JM, Jeon BH, Kim CD, et al: Apurinic/apyrimidinic endonuclease 1 is a key modulator of keratinocyte inflammatory responses. J Immunol. 183:6839–6848. 2009. View Article : Google Scholar : PubMed/NCBI
26	Xiong GS, Sun HL, Wu SM and Mo JZ: Small interfering RNA against the apurinic or apyrimidinic endonuclease enhances the sensitivity of human pancreatic cancer cells to gemcitabine in vitro. J Dig Dis. 11:224–230. 2010.PubMed/NCBI
27	Lou D, Zhu L, Ding H, Dai HY and Zou GM: Aberrant expression of redox protein Ape1 in colon cancer stem cells. Oncol Lett. 7:1078–1082. 2014.PubMed/NCBI
28	Funato Y, Michiue T, Asashima M and Miki H: The thioredoxin-related redox-regulating protein nucleoredoxin inhibits Wnt-beta-catenin signalling through dishevelled. Nat Cell Biol. 8:501–508. 2006. View Article : Google Scholar : PubMed/NCBI
29	Tetsu O and McCormick F: Beta-catenin regulates expression of cyclin D1 in colon carcinoma cells. Nature. 398:422–426. 1999. View Article : Google Scholar : PubMed/NCBI
30	Chen B, Dodge ME, Tang W, Lu J, Ma Z, Fan CW, Wei S, Hao W, Kilgore J, Williams NS, et al: Small molecule-mediated disruption of Wnt-dependent signaling in tissue regeneration and cancer. Nat Chem Biol. 5:100–107. 2009. View Article : Google Scholar : PubMed/NCBI
31	Barnes T, Kim WC, Mantha AK, Kim SE, Izumi T, Mitra S and Lee CH: Identification of Apurinic/apyrimidinic endonuclease 1 (APE1) as the endoribonuclease that cleaves c-myc mRNA. Nucleic Acids Res. 37:3946–3958. 2009. View Article : Google Scholar : PubMed/NCBI
32	Guttridge DC, Albanese C, Reuther JY, Pestell RG and Baldwin AS Jr: NF-kappaB controls cell growth and differentiation through transcriptional regulation of cyclin D1. Mol Cell Biol. 19:5785–5799. 1999.PubMed/NCBI
33	Wu HH, Cheng YW, Chang JT, Wu TC, Liu WS, Chen CY and Lee H: Subcellular localization of apurinic endonuclease 1 promotes lung tumor aggressiveness via NF-kappaB activation. Oncogene. 29:4330–4340. 2010. View Article : Google Scholar : PubMed/NCBI
34	Hariharan D, Saied A and Kocher HM: Analysis of mortality rates for pancreatic cancer across the world. HPB (Oxford). 10:58–62. 2008. View Article : Google Scholar
35	Kikuchi A, Kishida S and Yamamoto H: Regulation of Wnt signaling by protein-protein interaction and post-translational modifications. Exp Mol Med. 38:1–10. 2006. View Article : Google Scholar : PubMed/NCBI
36	Hoffmeyer K, Raggioli A, Rudloff S, Anton R, Hierholzer A, Del Valle I, Hein K, Vogt R and Kemler R: Wnt/β-catenin signaling regulates telomerase in stem cells and cancer cells. Science. 336:1549–1554. 2012. View Article : Google Scholar : PubMed/NCBI
37	Clevers H and Nusse R: Wnt/β-catenin signaling and disease. Cell. 149:1192–1205. 2012. View Article : Google Scholar : PubMed/NCBI
38	Sastre-Perona A and Santisteban P: Role of the wnt pathway in thyroid cancer. Front Endocrinol (Lausanne). 3:312012.
39	Zeng G, Germinaro M, Micsenyi A, Monga NK, Bell A, Sood A, Malhotra V, Sood N, Midda V, Monga DK, et al: Aberrant Wnt/beta-catenin signaling in pancreatic adenocarcinoma. Neoplasia. 8:279–289. 2006. View Article : Google Scholar : PubMed/NCBI
40	Yu M, Ting DT, Stott SL, Wittner BS, Ozsolak F, Paul S, Ciciliano JC, Smas ME, Winokur D, Gilman AJ, et al: RNA sequencing of pancreatic circulating tumour cells implicates WNT signalling in metastasis. Nature. 487:510–513. 2012. View Article : Google Scholar : PubMed/NCBI
41	Grek CL and Tew KD: Redox metabolism and malignancy. Curr Opin Pharmacol. 10:362–368. 2010. View Article : Google Scholar : PubMed/NCBI