Abstract
In studying the age dependence and chronology of ovarian tumors in follicle stimulating hormone receptor knockout mice, we identified a novel ovarian tumor associated gene-12 (OTAG-12), which is progressively downregulated and maps to Chr. 8B3.3. OTAG-12 protein overexpression in mouse ovarian and mammary tumor cells suggested powerful anti-proliferative effects. In human epithelial ovarian cancers (OCs) and OC cell lines, OTAG-12 mRNA expression is downregulated in comparison with normal ovaries. Cloning and identification revealed that human OTAG-12 mapping to gene-rich Chr. 19p13.12 is expressed in three spliced forms: hOTAG-12a, hOTAG-12b and hOTAG-12c, of which b is predominant in the normal ovary. Functionally active hOTAG-12b is a simple protein with no disulfide bonds and a nuclear localization signal is present in all variants. Transfection of OTAG-12 variants in OC and tumorigenic HEK293 cells confirmed nuclear localization. hOTAG-12b overexpression in OC and HEK293 cells effectively suppressed cell growth, anchorage-dependent and independent colony formation followed by apoptosis, whereas hOTAG-12a and hOTAG-12c had no such effects. Deletion mutants identified the critical importance of carboxyl terminus for hOTAG-12b function. Doxycycline-inducible growth inhibition of HEK293 cells by hOTAG-12a was associated with effects on G2 cell cycle arrest and apoptosis induction. hOTAG-12b expression rendered tumorigenic cells more sensitive to four apoptotic stimuli including etoposide—a topoisomerase-II inhibitor. Doxycycline-induced hOTAG-12b expression blocked xenograft tumor growth in nude mice, whereas hOTAG-12a was ineffective. Although p53-pathway-dependent apoptotic agents could upregulate endogenous hOTAG-12b and p53 in UCI-101/107 OC cells, hOTAG-12b could also induce apoptosis in p53-null and platinum-resistant SKOV3 OC cells and Doxycycline-induced hOTAG-12b did not alter p53. Further study showed that hOTAG-12b increases mRNAs of pro-apoptotic genes such as BAD, GADD45α and CIEDB, while inhibiting anti-apoptotic NAIP and Akt1 expression, suggesting that hOTAG-12b-induced apoptosis might be p53-independent. These results indicate that hOTAG-12b is a putative ovarian tumor suppressor gene warranting further studies.
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References
Al-Romaih K, Sadikovic B, Yoshimoto M, Wang Y, Zielenska M, Squire JA . (2008). Decitabine-induced demethylation of 5′ CpG island in GADD45A leads to apoptosis in osteosarcoma cells. Neoplasia 10: 471–480.
Allen S, Sotos J, Sylte MJ, Czuprynski CJ . (2001). Use of Hoechst 33342 staining to detect apoptotic changes in bovine mononuclear phagocytes infected with Mycobacterium avium subsp. paratuberculosis. Clin Diagn Lab Immunol 8: 460–464.
Aravindakshan J, Chen X, Sairam MR . (2006a). Differential expression of claudin family proteins in mouse ovarian serous papillary epithelial adenoma in aging FSH receptor-deficient mutants. Neoplasia 8: 984–994.
Aravindakshan J, Chen XL, Sairam MR . (2006b). Age-dependent bimodal GDNF regulation during ovarian tumorigenesis in follitropin receptor mutant mice. Biochem Biophys Res Commun 351: 507–513.
Aravindakshan J, Chen XL, Sairam MR . (2010). Chronology and complexities of ovarian tumorigenesis in FORKO mice: age-dependent gene alterations and progressive dysregulation of Major Histocompatibility Complex (MHC) Class I and II profiles. Mol Cell Endocrinol 329: 37–46.
Barnes BJ, Kellum MJ, Pinder KE, Frisancho JA, Pitha PM . (2003). Interferon regulatory factor 5, a novel mediator of cell cycle arrest and cell death. Cancer Res 63: 6424–6431.
Bast Jr RC, Hennessy B, Mills GB . (2009). The biology of ovarian cancer: new opportunities for translation. Nat Rev Cancer 9: 415–428.
Bignone PA, Lee KY, Liu Y, Emilion G, Finch J, Soosay AE et al. (2007). RPS6KA2, a putative tumour suppressor gene at 6q27 in sporadic epithelial ovarian cancer. Oncogene 26: 683–700.
Boisvert-Adamo K, Aplin AE . (2008). Mutant B-RAF mediates resistance to anoikis via Bad and Bim. Oncogene 27: 3301–3312.
Bruening W, Prowse AH, Schultz DC, Holgado-Madruga M, Wong A, Godwin AK . (1999). Expression of OVCA1, a candidate tumor suppressor, is reduced in tumors and inhibits growth of ovarian cancer cells. Cancer Res 59: 4973–4983.
Chen F, Zhang Z, Leonard SS, Shi X . (2001). Contrasting roles of NF-kappaB and JNK in arsenite-induced p53-independent expression of GADD45alpha. Oncogene 20: 3585–3589.
Chen X, Aravindakshan J, Yang Y, Sairam MR . (2007). Early alterations in ovarian surface epithelial cells and induction of ovarian epithelial tumors triggered by loss of FSH receptor. Neoplasia 9: 521–531.
Chen X, Aravindakshan J, Yang Y, Tiwari-Pandey R, Sairam MR . (2006). Aberrant expression of PDGF ligands and receptors in the tumor prone ovary of follitropin receptor knockout (FORKO) mouse. Carcinogenesis 27: 903–915.
Chen Z, Guo K, Toh SY, Zhou Z, Li P . (2000). Mitochondria localization and dimerization are required for CIDE-B to induce apoptosis. J Biol Chem 275: 22619–22622.
Corney DC, Flesken-Nikitin A, Choi J, Nikitin AY . (2008). Role of p53 and Rb in ovarian cancer. Adv Exp Med Biol 622: 99–117.
Dai Y, Liu M, Tang W, Li Y, Lian J, Lawrence TS et al. (2009). A Smac-mimetic sensitizes prostate cancer cells to TRAIL-induced apoptosis via modulating both IAPs and NF-κB. BMC Cancer 9: 392–406.
Danilovich N, Roy I, Sairam MR . (2001). Ovarian pathology and high incidence of sex cord tumors in follitropin receptor knockout (FORKO) mice. Endocrinology 142: 3673–3684.
Dievart A, Beaulieu N, Jolicoeur P . (1999). Involvement of Notch1 in the development of mouse mammary tumors. Oncogene 18: 5973–5981.
Dinulescu DM, Ince TA, Quade BJ, Shafer SA, Crowley D, Jacks T . (2005). Role of K-ras and Pten in the development of mouse models of endometriosis and endometrioid ovarian cancer. Nat Med 11: 63–70.
Dougherty MK, Morrison DK . (2004). Unlocking the code of 14-3-3. J Cell Sci 117: 1875–1884.
Fuchtner C, Emma DA, Manetta A, Gamboa G, Bernstein R, Liao SY . (1993). Characterization of a human ovarian carcinoma cell line: UCI 101. Gynecol Oncol 48: 203–209.
Gamboa G, Carpenter PM, Podnos YD, Dorion G, Iravani L, Bolton D et al. (1995). Characterization and development of UCI 107, a primary human ovarian carcinoma cell line. Gynecol Oncol 58: 336–343.
Grimwood J, Gordon LA, Olsen A, Terry A, Schmutz J, Lamerdin J et al. (2004). The DNA sequence and biology of human chromosome 19. Nature 428: 529–535.
Harkin DP, Bean JM, Miklos D, Song YH, Truong VB, Englert C et al. (1999). Induction of GADD45 and JNK/SAPK-dependent apoptosis following inducible expression of BRCA1. Cell 97: 575–586.
Jarboe EA, Folkins AK, Drapkin R, Ince TA, Agoston ES, Crum CP . (2008). Tubal and ovarian pathways to pelvic epithelial cancer: a pathological perspective. Histopathology 53: 127–138.
Jemal A, Siegel R, Ward E, Hao Y, Xu J, Thun MJ . (2009). Cancer statistics, 2009. CA Cancer J Clin 59: 225–249.
Johnson GL, Lapadat R . (2002). Mitogen-activated protein kinase pathways mediated by ERK, JNK, and p38 protein kinases. Science 298: 1911–1912.
Konstantinopoulos PA, Spentzos D, Cannistra SA . (2008). Gene-expression profiling in epithelial ovarian cancer. Nat Clin Pract Oncol 5: 577–587.
Liston P, Fong WG, Korneluk RG . (2003). The inhibitors of apoptosis: there is more to life than Bcl2. Oncogene 22: 8568–8580.
Lugovskoy AA, Zhou P, Chou JJ, McCarty JS, Li P, Wagner G . (1999). Solution structure of the CIDE-N domain of CIDE-B and a model for CIDE-N/CIDE-N interactions in the DNA fragmentation pathway of apoptosis. Cell 99: 747–755.
Luo RZ, Fang X, Marquez R, Liu SY, Mills GB, Liao WS et al. (2003). ARHI is a Ras-related small G-protein with a novel N-terminal extension that inhibits growth of ovarian and breast cancers. Oncogene 22: 2897–2909.
Maertens GN, El Messaoudi-Aubert S, Elderkin S, Hiom K, Peters G . (2010). Ubiquitin-specific proteases 7 and 11 modulate Polycomb regulation of the INK4a tumour suppressor. EMBO J 29: 2553–2565.
Nowak-Markwitz E, Jankowska A, Andrusiewicz M, Szczerba A . (2004). Expression of beta-human chorionic gonadotropin in ovarian cancer tissue. Eur J Gynaecol Oncol 25: 465–469.
O'Riordan MX, Bauler LD, Scott FL, Duckett CS . (2008). Inhibitor of apoptosis proteins in eukaryotic evolution and development: a model of thematic conservation. Dev Cell 15: 497–508.
Pal T, Permuth-Wey J, Betts JA, Krischer JP, Fiorica J, Arango H et al. (2005). BRCA1 and BRCA2 mutations account for a large proportion of ovarian carcinoma cases. Cancer 104: 2807–2816.
Park JT, Li M, Nakayama K, Mao TL, Davidson B, Zhang Z et al. (2006). Notch3 gene amplification in ovarian cancer. Cancer Res 66: 6312–6318.
Roby KF, Taylor CC, Sweetwood JP, Cheng Y, Pace JL, Tawfik O et al. (2000). Development of a syngeneic mouse model for events related to ovarian cancer. Carcinogenesis 21: 585–591.
Schwartz MF, Lee SJ, Duong JK, Eminaga S, Stern DF . (2003). FHA domain-mediated DNA checkpoint regulation of Rad53. Cell Cycle 2: 384–396.
Thyss R, Virolle V, Imbert V, Peyron JF, Aberdam D, Virolle T . (2005). NF-kappaB/Egr-1/Gadd45 are sequentially activated upon UVB irradiation to mediate epidermal cell death. EMBO J 24: 128–137.
Wagner MW, Li LS, Morales JC, Galindo CL, Garner HR, Bornmann WG et al. (2008). Role of c-Abl kinase in DNA mismatch repair-dependent G2 cell cycle checkpoint arrest responses. J Biol Chem 283: 21382–21393.
Willis SN, Adams JM . (2005). Life in the balance: how BH3-only proteins induce apoptosis. Curr Opin Cell Biol 17: 617–625.
Wu R, Hendrix-Lucas N, Kuick R, Zhai Y, Schwartz DR, Akyol A et al. (2007). Mouse model of human ovarian endometrioid adenocarcinoma based on somatic defects in the Wnt/beta-catenin and PI3K/Pten signaling pathways. Cancer Cell 11: 321–333.
Wu Z, Tandon R, Ziembicki J, Nagano J, Hujer KM, Miller RT et al. (2005). Role of ceramide in Ca2+-sensing receptor-induced apoptosis. J Lipid Res 46: 1396–1404.
Yan X, Fraser M, Qiu Q, Tsang BK . (2006). Over-expression of PTEN sensitizes human ovarian cancer cells to cisplatin-induced apoptosis in a p53-dependent manner. Gynecol Oncol 102: 348–355.
Yin BW, Dnistrian A, Lloyd KO . (2002). Ovarian cancer antigen CA125 is encoded by the MUC16 mucin gene. Int J Cancer 98: 737–740.
Zhu QS, Ren W, Korchin B, Lahat G, Dicker A, Lu Y et al. (2008). Soft tissue sarcoma cells are highly sensitive to AKT blockade: a role for p53-independent up-regulation of GADD45 alpha. Cancer Res 68: 2895–2903.
Acknowledgements
We are grateful to Dr P Carpenter (UC Irvine) for providing ovarian cancer cell lines UCI101 and UCI107, Dr Roby KF (Univ Kansas) for ID8 cells, Drs Jolicoeur for HC11-26 cells and Seidah at IRCM for the modified HEK293 cell lines. We appreciate the useful suggestions provided by Drs Zhongjun Dong and Hua Lin during this investigation. We also acknowledge the help of Drs Marie-Claude Beauchamp and Amber Yasmeen in this study. This investigation was supported in part by the NCIC and the CIHR.
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Chen, X., Zhang, H., Aravindakshan, J. et al. Anti-proliferative and pro-apoptotic actions of a novel human and mouse ovarian tumor-associated gene OTAG-12: downregulation, alternative splicing and drug sensitization. Oncogene 30, 2874–2887 (2011). https://doi.org/10.1038/onc.2011.11
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DOI: https://doi.org/10.1038/onc.2011.11
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