ISG15 inhibits cancer cell growth and promotes apoptosis

Zhou,Mei-Juan; Chen,Fang-Zhi; Chen,Han-Chun; Wan,Xin-Xing; Zhou,Xi; Fang,Qian; Zhang,Dian-Zheng

doi:10.3892/ijmm.2016.2845

ISG15 inhibits cancer cell growth and promotes apoptosis

Authors:
- Mei-Juan Zhou
- Fang-Zhi Chen
- Han-Chun Chen
- Xin-Xing Wan
- Xi Zhou
- Qian Fang
- Dian-Zheng Zhang
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Affiliations: Department of Biochemistry, School of Life Sciences and the State Key Laboratory of Medical Genetics, Central South University, Changsha, Hunan 410013, P.R. China, Department of Urology, The Second Xiangya Hospital of Central South University, Changsha, Hunan 410011, P.R. China
Published online on: December 30, 2016 https://doi.org/10.3892/ijmm.2016.2845
Pages: 446-452

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Abstract

Cervical cancer is one of the most common causes of cancer-related mortality in women in developing countries. Interferon (IFN)-α has been widely used in the treatment of various types of cancer, including cervical cancer, and IFN-stimulated gene 15 (ISG15), an ubiquitin-like protein, is upregulated by IFN-α treatment. The anti-virus and antitumor effects of ISG15 have been reported; however, its mechanism of action have not yet been fully elucidated. In this study, HeLa cells were used as a model system to investigate the roles of ISG15 in IFN-α-mediated cancer cell growth inhibition and induction of apoptosis. The results revealed that both p53 and p21 were upregulated in HeLa cells treated with IFN-α or in the HeLa cells overexpressing ISG15. In addition, the expression levels of ubiquitin-like modifier-activating enzyme 7 (UBA7, also known as UBE1L; ISG15 E1-activating enzyme), UBCH8 (ISG15 E2-conjugating enzyme) and HERC5 (ISG15 E3-ligase) were elevated in the HeLa cells treated with IFN-α. The levels of p53 in the HeLa cells were attenuated by transient transfection with small interfering RNA (siRNA) targeting ISG15 (ISG15-siRNA). Cell viability was inhibited by both IFN-α treatment and ISG15 overexpression. However, these effects were significantly diminished when p53 was knocked down, suggesting that the effects of inhibitory effects of ISG15 on HeLa cell growth and the induction of apoptosis were p53-dependent. Taken together, these results suggest the existence of the IFN-α/ISG15/p53 axis in cervical cancer cells and any strategies manipulating the levels of ISG15 may thus prove to be effective in the treatment of cervical cancer.

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1	Duenas-Gonzalez A, Serrano-Olvera A, Cetina L and Coronel J: New molecular targets against cervical cancer. Int J Womens Health. 6:1023–1031. 2014. View Article : Google Scholar : PubMed/NCBI
2	Dasari S, Wudayagiri R and Valluru L: Cervical cancer: biomarkers for diagnosis and treatment. Clin Chim Acta. 445:7–11. 2015. View Article : Google Scholar : PubMed/NCBI
3	Schiffman MH: Recent progress in defining the epidemiology of human papillomavirus infection and cervical neoplasia. J Natl Cancer Inst. 84:394–398. 1992. View Article : Google Scholar : PubMed/NCBI
4	Massad LS, Turyk ME, Bitterman P and Wilbanks GD: Interferon-alpha and all-trans-retinoic acid reversibly inhibit the in vitro proliferation of cell lines derived from cervical cancers. Gynecol Oncol. 60:428–434. 1996. View Article : Google Scholar : PubMed/NCBI
5	Kim KY, Blatt L and Taylor MW: The effects of interferon on the expression of human papillomavirus oncogenes. J Gen Virol. 81:695–700. 2000. View Article : Google Scholar : PubMed/NCBI
6	Koromilas AE, Li S and Matlashewski G: Control of interferon signaling in human papillomavirus infection. Cytokine Growth Factor Rev. 12:157–170. 2001. View Article : Google Scholar : PubMed/NCBI
7	Barnard P, Payne E and McMillan NA: The human papillomavirus E7 protein is able to inhibit the antiviral and anti-growth functions of interferon-α. Virology. 277:411–419. 2000. View Article : Google Scholar : PubMed/NCBI
8	Lenschow DJ, Lai C, Frias-Staheli N, Giannakopoulos NV, Lutz A, Wolff T, Osiak A, Levine B, Schmidt RE, García-Sastre A, et al: IFN-stimulated gene 15 functions as a critical antiviral molecule against influenza, herpes, and Sindbis viruses. Proc Natl Acad Sci USA. 104:1371–1376. 2007. View Article : Google Scholar : PubMed/NCBI
9	Friedman RM: Clinical uses of interferons. Br J Clin Pharmacol. 65:158–162. 2008. View Article : Google Scholar :
10	Sadler AJ and Williams BR: Interferon-inducible antiviral effectors. Nat Rev Immunol. 8:559–568. 2008. View Article : Google Scholar : PubMed/NCBI
11	Zhang T, Sun HC, Zhou HY, Luo JT, Zhang BL, Wang P, Wang L, Qin LX, Ren N, Ye SL, et al: Interferon alpha inhibits hepatocellular carcinoma growth through inducing apoptosis and interfering with adhesion of tumor endothelial cells. Cancer Lett. 290:204–210. 2010. View Article : Google Scholar
12	Shi WY, Cao C and Liu L: Interferon α induces the apoptosis of cervical cancer HeLa cells by activating both the intrinsic mitochondrial pathway and endoplasmic reticulum stress-induced pathway. Int J Mol Sci. 17:pii: E1832. 2016. View Article : Google Scholar
13	Kim KI, Yan M, Malakhova O, Luo JK, Shen MF, Zou W, de la Torre JC and Zhang DE: Ube1L and protein ISGylation are not essential for alpha/beta interferon signaling. Mol Cell Biol. 26:472–479. 2006. View Article : Google Scholar :
14	Hishiki T, Han Q, Arimoto K, Shimotohno K, Igarashi T, Vasudevan SG, Suzuki Y and Yamamoto N: Interferon-mediated ISG15 conjugation restricts dengue virus 2 replication. Biochem Biophys Res Commun. 448:95–100. 2014. View Article : Google Scholar : PubMed/NCBI
15	Loeb KR and Haas AL: The interferon-inducible 15-kDa ubiquitin homolog conjugates to intracellular proteins. J Biol Chem. 267:7806–7813. 1992.PubMed/NCBI
16	Haas AL, Ahrens P, Bright PM and Ankel H: Interferon induces a 15-kilodalton protein exhibiting marked homology to ubiquitin. J Biol Chem. 262:11315–11323. 1987.PubMed/NCBI
17	Darb-Esfahani S, Sinn BV, Rudl M, Sehouli J, Braicu I, Dietel M and Denkert C: Interferon-stimulated gene, 15 kDa (ISG15) in ovarian high-grade serous carcinoma: prognostic impact and link to NF-κB pathway. Int J Gynecol Pathol. 33:16–22. 2014. View Article : Google Scholar
18	Jeon YJ, Yoo HM and Chung CH: ISG15 and immune diseases. Biochim Biophys Acta. 1802:485–496. 2010. View Article : Google Scholar : PubMed/NCBI
19	Burks J, Reed RE and Desai SD: Free ISG15 triggers an antitumor immune response against breast cancer: a new perspective. Oncotarget. 6:7221–7231. 2015. View Article : Google Scholar : PubMed/NCBI
20	Zhao C, Hsiang TY, Kuo RL and Krug RM: ISG15 conjugation system targets the viral NS1 protein in influenza A virus-infected cells. Proc Natl Acad Sci USA. 107:2253–2258. 2010. View Article : Google Scholar : PubMed/NCBI
21	Jeon YJ, Jo MG, Yoo HM, Hong SH, Park JM, Ka SH, Oh KH, Seol JH, Jung YK and Chung CH: Chemosensitivity is controlled by p63 modification with ubiquitin-like protein ISG15. J Clin Invest. 122:2622–2636. 2012. View Article : Google Scholar : PubMed/NCBI
22	Wan XX, Chen HC, Khan MA, Xu AH, Yang FL, Zhang YY and Zhang DZ: ISG15 inhibits IFN-α-resistant liver cancer cell growth. BioMed Res Int. 2013:5709092013. View Article : Google Scholar
23	Strander H and Einhorn S: Interferons and the tumor cell. Biotherapy. 8:213–218. 1996. View Article : Google Scholar : PubMed/NCBI
24	Pfeffer LM, Dinarello CA, Herberman RB, Williams BR, Borden EC, Bordens R, Walter MR, Nagabhushan TL, Trotta PP and Pestka S: Biological properties of recombinant alpha-interferons: 40th anniversary of the discovery of interferons. Cancer Res. 58:2489–2499. 1998.PubMed/NCBI
25	Darnell JE Jr, Kerr IM and Stark GR: Jak-STAT pathways and transcriptional activation in response to IFNs and other extracellular signaling proteins. Science. 264:1415–1421. 1994. View Article : Google Scholar : PubMed/NCBI
26	O'Shea JJ, Gadina M and Schreiber RD: Cytokine signaling in 2002: new surprises in the Jak/Stat pathway. Cell. 109(Suppl): S121–S131. 2002. View Article : Google Scholar : PubMed/NCBI
27	Sainz B Jr, Martín B, Tatari M, Heeschen C and Guerra S: ISG15 is a critical microenvironmental factor for pancreatic cancer stem cells. Cancer Res. 74:7309–7320. 2014. View Article : Google Scholar : PubMed/NCBI
28	Takaoka A, Hayakawa S, Yanai H, Stoiber D, Negishi H, Kikuchi H, Sasaki S, Imai K, Shibue T, Honda K and Taniguchi T: Inte gration of interferon-α/β signalling to p53 responses in tumour suppression and antiviral defence. Nature. 424:516–523. 2003. View Article : Google Scholar : PubMed/NCBI
29	Malakhov MP, Malakhova OA, Kim KI, Ritchie KJ and Zhang DE: UBP43 (USP18) specifically removes ISG15 from conjugated proteins. J Biol Chem. 277:9976–9981. 2002. View Article : Google Scholar : PubMed/NCBI
30	Yuan W and Krug RM: Influenza B virus NS1 protein inhibits conjugation of the interferon (IFN)-induced ubiquitin-like ISG15 protein. EMBO J. 20:362–371. 2001. View Article : Google Scholar : PubMed/NCBI
31	D'Cunha J, Knight E Jr, Haas AL, Truitt RL and Borden EC: Immunoregulatory properties of ISG15, an interferon-induced cytokine. Proc Natl Acad Sci USA. 93:211–215. 1996. View Article : Google Scholar : PubMed/NCBI
32	Owhashi M, Taoka Y, Ishii K, Nakazawa S, Uemura H and Kambara H: Identification of a ubiquitin family protein as a novel neutrophil chemotactic factor. Biochem Biophys Res Commun. 309:533–539. 2003. View Article : Google Scholar : PubMed/NCBI
33	Campbell JA and Lenschow DJ: Emerging roles for immunomodulatory functions of free ISG15. J Interferon Cytokine Res. 33:728–738. 2013. View Article : Google Scholar : PubMed/NCBI
34	Desai SD, Reed RE, Burks J, Wood LM, Pullikuth AK, Haas AL, Liu LF, Breslin JW, Meiners S and Sankar S: ISG15 disrupts cytoskeletal architecture and promotes motility in human breast cancer cells. Exp Biol Med (Maywood). 237:38–49. 2012. View Article : Google Scholar
35	Burks J, Reed RE and Desai SD: ISGylation governs the oncogenic function of Ki-Ras in breast cancer. Oncogene. 33:794–803. 2014. View Article : Google Scholar
36	Kiessling A, Hogrefe C, Erb S, Bobach C, Fuessel S, Wessjohann L and Seliger B: Expression, regulation and function of the ISGylation system in prostate cancer. Oncogene. 28:2606–2620. 2009. View Article : Google Scholar : PubMed/NCBI
37	Yoshikawa A, Imagawa A, Nakata S, Fukui K, Kuroda Y, Miyata Y, Sato Y, Hanafusa T, Matsuoka TA, Kaneto H, et al: Interferon stimulated gene 15 has an anti-apoptotic effect on MIN6 cells. Endocr J. 61:883–890. 2014. View Article : Google Scholar : PubMed/NCBI
38	Butz K, Shahabeddin L, Geisen C, Spitkovsky D, Ullmann A and Hoppe-Seyler F: Functional p53 protein in human papillomavirus-positive cancer cells. Oncogene. 10:927–936. 1995.PubMed/NCBI
39	Mendonca MS, Howard KL, Farrington DL, Desmond LA, Temples TM, Mayhugh BM, Pink JJ and Boothman DA: Delayed apoptotic responses associated with radiation-induced neoplastic transformation of human hybrid cells. Cancer Res. 59:3972–3979. 1999.PubMed/NCBI
40	Yang J, Dai LX, Chen M, Li B, Ding N, Li G, Liu YQ, Li MY, Wang BN, Shi XL and Tan HB: Inhibition of antiviral drug cidofovir on proliferation of human papillomavirus-infected cervical cancer cells. Exp Ther Med. 12:2965–2973. 2016.PubMed/NCBI
41	Huang YF, Wee S, Gunaratne J, Lane DP and Bulavin DV: Isg15 controls p53 stability and functions. Cell Cycle. 13:2200–2210. 2014. View Article : Google Scholar : PubMed/NCBI
42	Huang YF and Bulavin DV: Oncogene-mediated regulation of p53 ISGylation and functions. Oncotarget. 5:5808–5818. 2014. View Article : Google Scholar : PubMed/NCBI
43	Menendez D and Anderson CW: p53 vs. ISG15: stop, you're killing me. Cell Cycle. 13:2160–2161. 2014. View Article : Google Scholar : PubMed/NCBI