Attenuation of MUC4 potentiates the anticancer activity of auranofin via regulation of the Her2/Akt/FOXO3 pathway in ovarian cancer cells

Bae,Jun  Sang; Lee,Jongsung; Park,Yoonkook; Park,Kyungmoon; Kim,Jung  Ryul; Cho,Dong  Hyu; Jang,Kyu  Yun; Park,See-Hyoung

doi:10.3892/or.2017.5853

Attenuation of MUC4 potentiates the anticancer activity of auranofin via regulation of the Her2/Akt/FOXO3 pathway in ovarian cancer cells

Authors:
- Jun Sang Bae
- Jongsung Lee
- Yoonkook Park
- Kyungmoon Park
- Jung Ryul Kim
- Dong Hyu Cho
- Kyu Yun Jang
- See-Hyoung Park
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Affiliations: Department of Pathology, Chonbuk National University Medical School, Research Institute of Clinical Medicine of Chonbuk National University-Biomedical Research Institute of Chonbuk National University Hospital and Research Institute for Endocrine Sciences, Jeonju, Republic of Korea, Department of Genetic Engineering, Sungkyunkwan University, Suwon, Republic of Korea, Department of Bio and Chemical Engineering, Hongik University, Sejong, Republic of Korea, Department of Orthopaedic Surgery, Chonbuk National University Medical School, Research Institute of Clinical Medicine of Chonbuk National University-Biomedical Research Institute of Chonbuk National University Hospital and Research Institute for Endocrine Sciences, Jeonju, Republic of Korea, Department of Obstetrics and Gynecology, Chonbuk National University Medical School, Research Institute of Clinical Medicine of Chonbuk National University- Biomedical Research Institute of Chonbuk National University Hospital, Jeonju, Republic of Korea
Published online on: July 28, 2017 https://doi.org/10.3892/or.2017.5853
Pages: 2417-2425

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Abstract

Previously, we reported that auranofin induces apoptosis in SKOV3 cells via regulation of the IKKβ/FOXO3 pathway. In the present study, we reveal that the anticancer activity of auranofin in SKOV3 cells could be enhanced by the attenuation of MUC4 through the regulation of the Her2/Akt/FOXO3 pathway. Compared to the control-siRNA, siRNA transfection against MUC4 into SKOV3 cells accelerated the protein degradation of Her2. Under the same conditions, the expression level of phosphorylated Akt was also downregulated leading to an increase of FOXO3 in the nucleus. Notably, auranofin treatment in SKOV3 cells also resulted in the downregulation of the expression levels of both Her2 and phosphorylated Akt. Thus, Her2 was identified as the common molecular target protein by siRNA transfection against MUC4. Western blot analysis of total and nuclear fraction lysates from SKOV3 cells revealed that attenuation of MUC4 combined with auranofin treatment in SKOV3 cells synergistically activated FOXO3 translocation from the cytoplasm to the nucleus through the regulation of the Her2/Akt/FOXO3 pathway. Attenuation of MUC4 by siRNA transfection potentiated the antitumor effect of auranofin which was examined by performing in vitro assays such as WST-1, cell counting, colony formation, TUNEL and Annexin V staining. In addition, western blot analysis of the apoptosis‑related proteins such as PARP1, caspase-3, Bim extra large (EL), Bax and Bcl2 revealed that the attenuation of MUC4 by siRNA transfection potentiates the pro-apoptotic activity of auranofin in SKOV3 cells. Collectively, auranofin could regulate the Her2/Akt/FOXO3 signaling pathway in SKOV3 cells and be used as a potential antitumor agent considering the expression of MUC4 in ovarian cancer patients.

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1	Hauber HP, Foley SC and Hamid Q: Mucin overproduction in chronic inflammatory lung disease. Can Respir J. 13:327–335. 2006. View Article : Google Scholar : PubMed/NCBI
2	Park HU, Kim JW, Kim GE, Bae HI, Crawley SC, Yang SC, Gum JR Jr, Batra SK, Rousseau K, Swallow DM, et al: Aberrant expression of MUC3 and MUC4 membrane-associated mucins and sialyl Le(x) antigen in pancreatic intraepithelial neoplasia. Pancreas. 26:e48–e54. 2003. View Article : Google Scholar : PubMed/NCBI
3	Singh AP, Chauhan SC, Bafna S, Johansson SL, Smith LM, Moniaux N, Lin MF and Batra SK: Aberrant expression of transmembrane mucins, MUC1 and MUC4, in human prostate carcinomas. Prostate. 66:421–429. 2006. View Article : Google Scholar : PubMed/NCBI
4	Voynow JA and Rubin BK: Mucins, mucus, and sputum. Chest. 135:505–512. 2009. View Article : Google Scholar : PubMed/NCBI
5	Rachagani S, Torres MP, Moniaux N and Batra SK: Current status of mucins in the diagnosis and therapy of cancer. Biofactors. 35:509–527. 2009. View Article : Google Scholar : PubMed/NCBI
6	Kaur S, Sharma N, Krishn SR, Lakshmanan I, Rachagani S, Baine MJ, Smith LM, Lele SM, Sasson AR, Guha S, et al: MUC4-mediated regulation of acute phase protein lipocalin 2 through HER2/AKT/NF-κB signaling in pancreatic cancer. Clin Cancer Res. 20:688–700. 2014. View Article : Google Scholar : PubMed/NCBI
7	Chaturvedi P, Singh AP, Moniaux N, Senapati S, Chakraborty S, Meza JL and Batra SK: MUC4 mucin potentiates pancreatic tumor cell proliferation, survival, and invasive properties and interferes with its interaction to extracellular matrix proteins. Mol Cancer Res. 5:309–320. 2007. View Article : Google Scholar : PubMed/NCBI
8	Bafna S, Kaur S and Batra SK: Membrane-bound mucins: The mechanistic basis for alterations in the growth and survival of cancer cells. Oncogene. 29:2893–2904. 2010. View Article : Google Scholar : PubMed/NCBI
9	Inata J, Hattori N, Yokoyama A, Ohshimo S, Doi M, Ishikawa N, Hamada H and Kohno N: Circulating KL-6/MUC1 mucin carrying sialyl Lewis^a oligosaccharide is an independent prognostic factor in patients with lung adenocarcinoma. Int J Cancer. 120:2643–2649. 2007. View Article : Google Scholar : PubMed/NCBI
10	Schroeder JA, Masri AA, Adriance MC, Tessier JC, Kotlarczyk KL, Thompson MC and Gendler SJ: MUC1 overexpression results in mammary gland tumorigenesis and prolonged alveolar differentiation. Oncogene. 23:5739–5747. 2004. View Article : Google Scholar : PubMed/NCBI
11	Elazeez Abd TA, El-Balshy A-L, Khalil MM, El-Tabye MM and Abdul-Halim H: Prognostic significance of P27 (Kip 1) and MUC1 in papillary transitional cell carcinoma of the urinary bladder. Urol Ann. 3:8–13. 2011. View Article : Google Scholar : PubMed/NCBI
12	Patriarca C, Colombo P, Taronna Pio A, Wesseling J, Franchi G, Guddo F, Naspro R, Macchi RM, Giunta P, Di Pasquale M, et al: Cell discohesion and multifocality of carcinoma in situ of the bladder: New insight from the adhesion molecule profile (e-cadherin, Ep-CAM, and MUC1). Int J Surg Pathol. 17:99–106. 2009. View Article : Google Scholar : PubMed/NCBI
13	Du L, Qian X, Dai C, Wang L, Huang D, Wang S and Shen X: Screening the molecular targets of ovarian cancer based on bioinformatics analysis. Tumori. 101:384–389. 2015. View Article : Google Scholar : PubMed/NCBI
14	Slichenmyer WJ and Fry DW: Anticancer therapy targeting the erbB family of receptor tyrosine kinases. Semin Oncol. 28 Suppl 16:S67–S79. 2001. View Article : Google Scholar
15	Schmidt M, Lewark B, Kohlschmidt N, Glawatz C, Steiner E, Tanner B, Pilch H, Weikel W, Kölbl H and Lehr HA: Long-term prognostic significance of HER-2/neu in untreated node-negative breast cancer depends on the method of testing. Breast Cancer Res. 7:R256–R266. 2005. View Article : Google Scholar : PubMed/NCBI
16	Ross JS and Fletcher JA: The HER-2/neu oncogene in breast cancer: Prognostic factor, predictive factor, and target for therapy. Oncologist. 3:237–252. 1998.PubMed/NCBI
17	Ladjemi MZ, Jacot W, Chardès T, Pèlegrin A and Navarro-Teulon I: Anti-HER2 vaccines: New prospects for breast cancer therapy. Cancer Immunol Immunother. 59:1295–1312. 2010. View Article : Google Scholar : PubMed/NCBI
18	Hynes NE and Stern DF: The biology of erbB-2/neu/HER-2 and its role in cancer. Biochim Biophys Acta. 1198:165–184. 1994.PubMed/NCBI
19	Ponnusamy MP, Seshacharyulu P, Vaz A, Dey P and Batra SK: MUC4 stabilizes HER2 expression and maintains the cancer stem cell population in ovarian cancer cells. J Ovarian Res. 4:72011. View Article : Google Scholar : PubMed/NCBI
20	Ponnusamy MP, Singh AP, Jain M, Chakraborty S, Moniaux N and Batra SK: MUC4 activates HER2 signalling and enhances the motility of human ovarian cancer cells. Br J Cancer. 99:520–526. 2008. View Article : Google Scholar : PubMed/NCBI
21	Park SH, Lee JH, Berek JS and Hu MC: Auranofin displays anticancer activity against ovarian cancer cells through FOXO3 activation independent of p53. Int J Oncol. 45:1691–1698. 2014. View Article : Google Scholar : PubMed/NCBI
22	Hu W, Li F, Mahavadi S and Murthy KS: Upregulation of RGS4 expression by IL-1beta in colonic smooth muscle is enhanced by ERK1/2 and p38 MAPK and inhibited by the PI3K/Akt/GSK3beta pathway. Am J Physiol Cell Physiol. 296:C1310–C1320. 2009. View Article : Google Scholar : PubMed/NCBI
23	Fu Z and Tindall DJ: FOXOs, cancer and regulation of apoptosis. Oncogene. 27:2312–2319. 2008. View Article : Google Scholar : PubMed/NCBI
24	Siegel R, Naishadham D and Jemal A: Cancer statistics, 2012. CA Cancer J Clin. 62:10–29. 2012. View Article : Google Scholar : PubMed/NCBI
25	Banerjee S and Kaye SB: New strategies in the treatment of ovarian cancer: Current clinical perspectives and future potential. Clin Cancer Res. 19:961–968. 2013. View Article : Google Scholar : PubMed/NCBI
26	Shigetomi H, Higashiura Y, Kajihara H and Kobayashi H: Targeted molecular therapies for ovarian cancer: An update and future perspectives (Review). Oncol Rep. 28:395–408. 2012.PubMed/NCBI
27	Singh AP, Moniaux N, Chauhan SC, Meza JL and Batra SK: Inhibition of MUC4 expression suppresses pancreatic tumor cell growth and metastasis. Cancer Res. 64:622–630. 2004. View Article : Google Scholar : PubMed/NCBI
28	Carraway KL, Perez A, Idris N, Jepson S, Arango M, Komatsu M, Haq B, Price-Schiavi SA, Zhang J and Carraway CA: Muc4/sialomucin complex, the intramembrane ErbB2 ligand, in cancer and epithelia: To protect and to survive. Prog Nucleic Acid Res Mol Biol. 71:149–185. 2002. View Article : Google Scholar : PubMed/NCBI
29	Chaturvedi P, Singh AP, Chakraborty S, Chauhan SC, Bafna S, Meza JL, Singh PK, Hollingsworth MA, Mehta PP and Batra SK: MUC4 mucin interacts with and stabilizes the HER2 oncoprotein in human pancreatic cancer cells. Cancer Res. 68:2065–2070. 2008. View Article : Google Scholar : PubMed/NCBI
30	Ramsauer VP, Pino V, Farooq A, Carraway Carothers CA, Salas PJ and Carraway KL: Muc4-ErbB2 complex formation and signaling in polarized CACO-2 epithelial cells indicate that Muc4 acts as an unorthodox ligand for ErbB2. Mol Biol Cell. 17:2931–2941. 2006. View Article : Google Scholar : PubMed/NCBI
31	Holbro T and Hynes NE: ErbB receptors: Directing key signaling networks throughout life. Annu Rev Pharmacol Toxicol. 44:195–217. 2004. View Article : Google Scholar : PubMed/NCBI
32	Katoh M and Katoh M: Human FOX gene family (Review). Int J Oncol. 25:1495–1500. 2004.PubMed/NCBI
33	Alvarez B, Martínez-A C, Burgering BM and Carrera AC: Forkhead transcription factors contribute to execution of the mitotic programme in mammals. Nature. 413:744–747. 2001. View Article : Google Scholar : PubMed/NCBI
34	Furukawa-Hibi Y, Kobayashi Y, Chen C and Motoyama N: FOXO transcription factors in cell-cycle regulation and the response to oxidative stress. Antioxid Redox Signal. 7:752–760. 2005. View Article : Google Scholar : PubMed/NCBI
35	Lu M, Zhao Y, Xu F, Wang Y, Xiang J and Chen D: The expression and prognosis of FOXO3a and Skp2 in human ovarian cancer. Med Oncol. 29:3409–3415. 2012. View Article : Google Scholar : PubMed/NCBI
36	Fei M, Zhao Y, Wang Y, Lu M, Cheng C, Huang X, Zhang D, Lu J, He S and Shen A: Low expression of Foxo3a is associated with poor prognosis in ovarian cancer patients. Cancer Invest. 27:52–59. 2009. View Article : Google Scholar : PubMed/NCBI
37	Park SH, Chung YM, Ma J, Yang Q, Berek JS and Hu MC: Pharmacological activation of FOXO3 suppresses triple-negative breast cancer in vitro and in vivo. Oncotarget. 7:42110–42125. 2016. View Article : Google Scholar : PubMed/NCBI
38	Gandin V, Fernandes AP, Rigobello MP, Dani B, Sorrentino F, Tisato F, Björnstedt M, Bindoli A, Sturaro A, Rella R, et al: Cancer cell death induced by phosphine gold(I) compounds targeting thioredoxin reductase. Biochem Pharmacol. 79:90–101. 2010. View Article : Google Scholar : PubMed/NCBI
39	Schuh E, Pflüger C, Citta A, Folda A, Rigobello MP, Bindoli A, Casini A and Mohr F: Gold(I) carbene complexes causing thioredoxin 1 and thioredoxin 2 oxidation as potential anticancer agents. J Med Chem. 55:5518–5528. 2012. View Article : Google Scholar : PubMed/NCBI
40	Pessetto ZY, Weir SJ, Sethi G, Broward MA and Godwin AK: Drug repurposing for gastrointestinal stromal tumor. Mol Cancer Ther. 12:1299–1309. 2013. View Article : Google Scholar : PubMed/NCBI
41	Li H, Hu J, Wu S, Wang L, Cao X, Zhang X, Dai B, Cao M, Shao R, Zhang R, et al: Auranofin-mediated inhibition of PI3K/AKT/mTOR axis and anticancer activity in non-small cell lung cancer cells. Oncotarget. 7:3548–3558. 2016. View Article : Google Scholar : PubMed/NCBI
42	Tanaka H, Fujita N and Tsuruo T: 3-Phosphoinositide-dependent protein kinase-1-mediated IkappaB kinase beta (IkkB) phosphorylation activates NF-kappaB signaling. J Biol Chem. 280:40965–40973. 2005. View Article : Google Scholar : PubMed/NCBI
43	Lee J, Bartholomeusz C, Mansour O, Humphries J, Hortobagyi GN, Ordentlich P and Ueno NT: A class I histone deacetylase inhibitor, entinostat, enhances lapatinib efficacy in HER2-overexpressing breast cancer cells through FOXO3-mediated Bim1 expression. Breast Cancer Res Treat. 146:259–272. 2014. View Article : Google Scholar : PubMed/NCBI