STAT1‑HDAC4 signaling induces epithelial‑mesenchymal transition and sphere formation of cancer cells overexpressing the oncogene, CUG2

Kaowinn,Sirichat; Kaewpiboon,Chutima; Koh,Sang Seok; Krämer,Oliver H.; Chung,Young‑Hwa

doi:10.3892/or.2018.6701

STAT1‑HDAC4 signaling induces epithelial‑mesenchymal transition and sphere formation of cancer cells overexpressing the oncogene, CUG2

Authors:
- Sirichat Kaowinn
- Chutima Kaewpiboon
- Sang Seok Koh
- Oliver H. Krämer
- Young‑Hwa Chung
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Affiliations: BK21+, Department of Cogno‑Mechatronics Engineering, Pusan National University, Busan 46241, Republic of Korea, Department of Biology, Faculty of Science, Thaksin University, Phatthalung 93210, Thailand, Department of Biological Sciences, Dong‑A University, Busan 49315, Republic of Korea, Department of Toxicology, University Medical Center Mainz, Mainz D‑55131, Germany
Published online on: September 12, 2018 https://doi.org/10.3892/or.2018.6701
Pages: 2619-2627
Copyright: © Kaowinn et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

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Abstract

Our previous studies have shown that the novel oncogene, cancer upregulated gene 2 (CUG2), activates STAT1, which is linked to anticancer drug resistance, induces epithelial‑mesenchymal transition (EMT) and cancer stem cell‑like phenotypes as determined by MTT, migration and sphere formation assays. We thus aimed to ascertain whether the activation of STAT1 by CUG2 is involved in these malignant phenotypes besides drug resistance. Here, we showed that STAT1 suppression decreased the expression of N‑cadherin and vimentin, biomarkers of EMT, which led to inhibition of the migration and invasion of human lung A549 cancer cells stably expressing CUG2, but did not recover E‑cadherin expression. STAT1 siRNA also diminished CUG2‑induced TGF‑β signaling, which is critical in EMT, and TGF‑β transcriptional activity. Conversely, inhibition of TGF‑β signaling reduced phosphorylation of STAT1, indicating a crosstalk between STAT1 and TGF‑β signaling. Furthermore, STAT1 silencing diminished sphere formation, which was supported by downregulation of stemness‑related factors such as Sox2, Oct4, and Nanog. Constitutive suppression of STAT1 also inhibited cell migration, invasion and sphere formation. As STAT1 acetylation counteracts STAT1 phosphorylation, acetylation of STAT1 by treatment with trichostatin A, an inhibitor of histone deacetylases (HDACs), reduced cell migration, invasion, and sphere formation. As HDAC4 is known to target STAT1, its role was investigated under CUG2 overexpression. HDAC4 suppression resulted in inhibition of cell migration, invasion, and sphere formation as HDAC4 silencing hindered TGF‑β signaling and decreased expression of Sox2 and Nanog. Taken together, we suggest that STAT1‑HDAC4 signaling induces malignant tumor features such as EMT and sphere formation in CUG2‑overexpressing cancer cells.

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1	Lee S, Gang J, Jeon SB, Choo SH, Lee B, Kim YG, Lee YS, Jung J, Song SY and Koh SS: Molecular cloning and functional analysis of a novel oncogene, cancer-upregulated gene 2 (CUG2). Biochem Biophys Res Commun. 360:633–639. 2007. View Article : Google Scholar : PubMed/NCBI
2	Hori T, Amano M, Suzuki A, Backer CB, Welburn JP, Dong Y, McEwen BF, Shang WH, Suzuki E, Okawa K, et al: CCAN makes multiple contacts with centromeric DNA to provide distinct pathways to the outer kinetochore. Cell. 135:1039–1052. 2008. View Article : Google Scholar : PubMed/NCBI
3	Kim H, Lee M and Lee S, Park B, Koh W, Lee DJ, Lim DS and Lee S: Cancer-upregulated gene 2 (CUG2), a new component of centromere complex, is required for kinetochore function. Mol Cells. 27:697–701. 2009. View Article : Google Scholar : PubMed/NCBI
4	Park EH, Park EH, Cho IR, Srisuttee R, Min HJ, Oh MJ, Jeong YJ, Jhun BH, Johnston RN, Lee S, et al: CUG2, a novel oncogene confers reoviral replication through Ras and p38 signaling pathway. Cancer Gene Ther. 17:307–314. 2010. View Article : Google Scholar : PubMed/NCBI
5	McEntee G, Kyula JN, Mansfield D, Smith H, Wilkinson M, Gregory C, Roulstone V, Coffey M and Harrington KJ: Enhanced cytotoxicity of reovirus and radiotherapy in melanoma cells is mediated through increased viral replication and mitochondrial apoptotic signalling. Oncotarget. 7:48517–48532. 2016. View Article : Google Scholar : PubMed/NCBI
6	Sborov DW, Nuovo GJ, Stiff A, Mace T, Lesinski GB, Benson DM Jr, Efebera YA, Rosko AE, Pichiorri F, Grever MR, et al: A phase I trial of single-agent reolysin in patients with relapsed multiple myeloma. Clin Cancer Res. 20:5946–5955. 2014. View Article : Google Scholar : PubMed/NCBI
7	Malilas W, Koh SS, Srisuttee R, Boonying W, Cho IR, Jeong CS, Johnston RN and Chung YH: Cancer upregulated gene 2, a novel oncogene, confers resistance to oncolytic vesicular stomatitis virus through STAT1-OASL2 signaling. Cancer Gene Ther. 20:125–132. 2013. View Article : Google Scholar : PubMed/NCBI
8	Malilas W, Koh SS, Kim S, Srisuttee R, Cho IR, Moon J, Yoo HS, Oh S, Johnston RN and Chung YH: Cancer upregulated gene 2, a novel oncogene, enhances migration and drug resistance of colon cancer cells via STAT1 activation. Int J Oncol. 43:1111–1116. 2013. View Article : Google Scholar : PubMed/NCBI
9	Kaowinn S, Kim J, Lee J, Shin DH, Kang CD, Kim DK, Lee S, Kang MK, Koh SS, Kim SJ, et al: Cancer upregulated gene 2 induces epithelial-mesenchymal transition of human lung cancer cells via TGF-βeta signaling. Oncotarget. 8:5092–5110. 2017. View Article : Google Scholar : PubMed/NCBI
10	Chin YE, Kitagawa M, Kuida K, Flavell RA and Fu XY: Activation of the STAT signaling pathway can cause expression of caspase 1 and apoptosis. Mol Cell Biol. 17:5328–5337. 1997. View Article : Google Scholar : PubMed/NCBI
11	Kumar A, Commane M, Flickinger TW, Horvath CM and Stark GR: Defective TNF-alpha-induced apoptosis in STAT1-null cells due to low constitutive levels of caspases. Science. 278:1630–1632. 1997. View Article : Google Scholar : PubMed/NCBI
12	Chin YE, Kitagawa M, Su WC, You ZH, Iwamoto Y and Fu XY: Cell growth arrest and induction of cyclin-dependent kinase inhibitor p21^WAF1/CIP1 mediated by STAT1. Science. 272:719–722. 1996. View Article : Google Scholar : PubMed/NCBI
13	Townsend PA, Scarabelli TM, Davidson SM, Knight RA, Latchman DS and Stephanou A: STAT-1 interacts with p53 to enhance DNA damage-induced apoptosis. J Biol Chem. 279:5811–5820. 2004. View Article : Google Scholar : PubMed/NCBI
14	Khodarev NN, Beckett M, Labay E, Darga T, Roizman B and Weichselbaum RR: STAT1 is overexpressed in tumors selected for radioresistance and confers protection from radiation in transduced sensitive cells. Proc Natl Acad Sci USA. 101:1714–1719. 2004. View Article : Google Scholar : PubMed/NCBI
15	Weichselbaum RR, Ishwaran H, Yoon T, Nuyten DS, Baker SW, Khodarev N, Su AW, Shaikh AY, Roach P, Kreike B, et al: An interferon-related gene signature for DNA damage resistance is a predictive marker for chemotherapy and radiation for breast cancer. Proc Natl Acad Sci USA. 105:18490–18495. 2008. View Article : Google Scholar : PubMed/NCBI
16	Fryknas M, Dhar S, Oberg F, Rickardson L, Rydaker M, Goransson H, Gustafsson M, Pettersson U, Nygren P, Larsson R and Isaksson A: STAT1 signaling is associated with acquired crossresistance to doxorubicin and radiation in myeloma cell lines. Int J Cancer. 120:189–195. 2007. View Article : Google Scholar : PubMed/NCBI
17	Roberts D, Schick J, Conway S, Biade S, Laub PB, Stevenson JP, Hamilton TC, O'Dwyer PJ and Johnson SW: Identification of genes associated with platinum drug sensitivity and resistance in human ovarian cancer cells. Br J Cancer. 92:1149–1158. 2005. View Article : Google Scholar : PubMed/NCBI
18	Yang XJ and Seto E: HATs and HDACs: From structure, function and regulation to novel strategies for therapy and prevention. Oncogene. 26:5310–5318. 2007. View Article : Google Scholar : PubMed/NCBI
19	Geng H, Harvey CT, Pittsenbarger J, Liu Q, Beer TM, Xue C and Qian DZ: HDAC4 protein regulates HIF1α protein lysine acetylation and cancer cell response to hypoxia. J Biol Chem. 286:38095–38102. 2011. View Article : Google Scholar : PubMed/NCBI
20	Mihaylova MM, Vasquez DS, Ravnskjaer K, Denechaud PD, Yu RT, Alvarez JG, Downes M, Evans RM, Montminy M and Shaw RJ: Class IIa histone deacetylases are hormone-activated regulators of FOXO and mammalian glucose homeostasis. Cell. 145:607–621. 2011. View Article : Google Scholar : PubMed/NCBI
21	Stronach EA, Alfraidi A, Rama N, Datler C, Studd JB, Agarwal R, Guney TG, Gourley C, Hennessy BT, Mills GB, et al: HDAC4-regulated STAT1 activation mediates platinum resistance in ovarian cancer. Cancer Res. 71:4412–4422. 2011. View Article : Google Scholar : PubMed/NCBI
22	Giaginis C, Alexandrou P, Delladetsima I, Giannopoulou I, Patsouris E and Theocharis S: Clinical significance of histone deacetylase (HDAC)-1, HDAC-2, HDAC-4, and HDAC-6 expression in human malignant and benign thyroid lesions. Tumour Biol. 35:61–71. 2014. View Article : Google Scholar : PubMed/NCBI
23	Wilson AJ, Byun DS, Nasser S, Murray LB, Ayyanar K, Arango D, Figueroa M, Melnick A, Kao GD, Augenlicht LH and Mariadason JM: HDAC4 promotes growth of colon cancer cells via repression of p21. Mol Biol Cell. 19:4062–4075. 2008. View Article : Google Scholar : PubMed/NCBI
24	Shen YF, Wei AM, Kou Q, Zhu QY and Zhang L: Histone deacetylase 4 increases progressive epithelial ovarian cancer cells via repression of p21 on fibrillar collagen matrices. Oncol Rep. 35:948–954. 2016. View Article : Google Scholar : PubMed/NCBI
25	Kang ZH, Wang CY, Zhang WL, Zhang JT, Yuan CH, Zhao PW, Lin YY, Hong S, Li CY and Wang L: Histone deacetylase HDAC4 promotes gastric cancer SGC-7901 cells progression via p21 repression. PLoS One. 9:e988942014. View Article : Google Scholar : PubMed/NCBI
26	Kim SJ, Glick A, Sporn MB and Roberts AB: Characterization of the promoter region of the human transforming growth factor-beta 1 gene. J Biol Chem. 264:402–408. 1989.PubMed/NCBI
27	Zhang Y, Wang W, Wang Y, Huang X, Zhang Z, Chen B, Xie W, Li S, Shen S and Peng B: NEK2 promotes hepatocellular carcinoma migration and invasion through modulation of the epithelial-mesenchymal transition. Oncol Rep. 39:1023–1033. 2018.PubMed/NCBI
28	Kramer OH and Heinzel T: Phosphorylation-acetylation switch in the regulation of STAT1 signaling. Mol Cell Endocrinol. 315:40–48. 2010. View Article : Google Scholar : PubMed/NCBI
29	Jechlinger M, Sommer A, Moriggl R, Seither P, Kraut N, Capodiecci P, Donovan M, Cordon-Cardo C, Beug H and Grunert S: Autocrine PDGFR signaling promotes mammary cancer metastasis. J Clin Invest. 116:1561–1570. 2006. View Article : Google Scholar : PubMed/NCBI
30	Seshacharyulu P, Ponnusamy MP, Rachagani S, Lakshmanan I, Haridas D, Yan Y, Ganti AK and Batra SK: Targeting EGF-receptor(s)-STAT1 axis attenuates tumor growth and metastasis through downregulation of MUC4 mucin in human pancreatic cancer. Oncotarget. 6:5164–5181. 2015. View Article : Google Scholar : PubMed/NCBI
31	Suyama K, Onishi H, Imaizumi A, Shinkai K, Umebayashi M, Kubo M, Mizuuchi Y, Oda Y, Tanaka M, Nakamura M, et al: CD24 suppresses malignant phenotype by downregulation of SHH transcription through STAT1 inhibition in breast cancer cells. Cancer Lett. 374:44–53. 2016. View Article : Google Scholar : PubMed/NCBI
32	Zeng LS, Yang XZ, Wen YF, Mail SJ, Wang MH, Zhang MY, Zheng XF and Wang HY: Overexpressed HDAC4 is associated with poor survival and promotes tumor progression in esophageal carcinoma. Aging. 8:1236–1249. 2016. View Article : Google Scholar : PubMed/NCBI
33	Kikuchi S, Suzuki R, Ohguchi H, Yoshida Y, Lu D, Cottini F, Jakubikova J, Bianchi G, Harada T, Gorgun G, et al: Class IIa HDAC inhibition enhances ER stress-mediated cell death in multiple myeloma. Leukemia. 29:1918–1927. 2015. View Article : Google Scholar : PubMed/NCBI
34	Marek L, Hamacher A, Hansen FK, Kuna K, Gohlke H, Kassack MU and Kurz T: Histone deacetylase (HDAC) inhibitors with a novel connecting unit linker region reveal a selectivity profile for HDAC4 and HDAC5 with improved activity against chemoresistant cancer cells. J Med Chem. 56:427–436. 2013. View Article : Google Scholar : PubMed/NCBI
35	Kaowinn S, Jun SW, Kim CS, Shin DM, Hwang YH, Kim K, Shin B, Kaewpiboon C, Jeong HH, Koh SS, et al: Increased EGFR expression induced by a novel oncogene, CUG2, confers resistance to doxorubicin through Stat1-HDAC4 signaling. Cell Oncol. 40:549–561. 2017. View Article : Google Scholar
36	Kramer OH, Knauer SK, Greiner G, Jandt E, Reichardt S, Gührs KH, Stauber RH, Böhmer FD and Heinzel T: A phosphorylation-acetylation switch regulates STAT1 signaling. Genes Dev. 23:223–235. 2009. View Article : Google Scholar : PubMed/NCBI
37	Ginter T, Bier C, Knauer SK, Sughra K, Hildebrand D, Munz T, Liebe T, Heller R, Henke A, Stauber RH, et al: Histone deacetylase inhibitors block IFNγ-induced STAT1 phosphorylation. Cell Signal. 24:1453–1460. 2012. View Article : Google Scholar : PubMed/NCBI