Effects of Treg cells and IDO on human epithelial ovarian cancer cells under hypoxic conditions

Liu,Jun; Zhang,Haiyan; Jia,Luoqi; Sun,Hong

doi:10.3892/mmr.2014.2893

Effects of Treg cells and IDO on human epithelial ovarian cancer cells under hypoxic conditions

Authors:
- Jun Liu
- Haiyan Zhang
- Luoqi Jia
- Hong Sun
View Affiliations

Affiliations: Department of Gynecology, Obstetrics and Gynecology Hospital, Fudan University, Shanghai 200090, P.R. China
Published online on: November 7, 2014 https://doi.org/10.3892/mmr.2014.2893
Pages: 1708-1714
Copyright: © Liu et al. This is an open access article distributed under the terms of Creative Commons Attribution License [CC BY_NC 3.0].

Metrics: Total Views: 0 (Spandidos Publications: | PMC Statistics: )

Metrics: Total PDF Downloads: 0 (Spandidos Publications: | PMC Statistics: )

Cited By (CrossRef): 0 citations Loading Articles...

Abstract

The aim of the present study was to explore the effect of hypoxia on ovarian cancer. A total of 6 samples were analyzed: SKOV3‑IP cells (ovarian cancer cell line); SKOV3‑IP and regulatory T (Treg) cells; SKOV3‑IP and cytotoxic T lymphocytes (CTLs); SKOV3‑IP and natural killer (NK) cells; SKOV3‑IP co-cultured with CTLs and Treg cells; and SKOV3‑IP co-cultured with Treg cells and NK cells. The expression of indoleamine 2,3‑dioxygenase (IDO) was detected by reverse transcription-polymerase chain reaction (RT‑PCR) and western blot analysis. An enzyme‑linked immunosorbent assay (ELISA) was used to detect the concentration of transforming growth factor‑β (TGF‑β), interferon‑γ (IFN‑γ), interleukin‑2 (IL‑2), interleukin‑10 (IL‑10), and perforin. Moreover, ovarian cancer cell apoptosis and invasive ability were examined using flow cytometry and a Transwell chamber assay. IDO expression was significantly reduced in hypoxia and enhanced by Treg cells. Treg cells inhibited the IL‑2, IFN‑γ and perforin secretion, and significantly (P<0.05) increased the IL‑10 and TGF‑β levels. The effects of Treg cells were enhanced with prolongation of the cell exposure to hypoxic conditions. In addition, Treg cells attenuated the promotive effect of CTLs and NK cells on cancer cell apoptosis. In addition, Treg cells significantly increased the SKOV3‑IP invasive ability (P=0.00109) under hypoxic conditions. Our results suggest that IDO and Treg cells may serve as important therapeutic targets for patients with ovarian cancer.

View Figures

View References

1	Fehrmann RS, Li XY, van der Zee AG, et al: Profiling studies in ovarian cancer: a review. Oncologist. 12:960–966. 2007. View Article : Google Scholar : PubMed/NCBI
2	Dougan M and Dranoff G: Immunotherapy of cancer. Innate Immune Regulation and Cancer Immunotherapy. Wang RF: Springer; New York: pp. 391–414. 2012, View Article : Google Scholar
3	Wållberg M, Wong FS and Green EA: An islet-specific pulse of TGF-β abrogates CTL function and promotes β cell survival independent of Foxp3⁺ T cells. J Immunol. 186:2543–2551. 2011. View Article : Google Scholar
4	Lu L, Zhou X, Wang J, Zheng SG and Horwitz DA: Characterization of protective human CD4⁺CD25⁺ FOXP3⁺ regulatory T cells generated with IL-2, TGF-β and retinoic acid. PLoS One. 5:e151502010. View Article : Google Scholar
5	Teoh D and Secord AA: Antiangiogenic agents in combination with chemotherapy for the treatment of epithelial ovarian cancer. Int J Gynecol Cancer. 22:348–359. 2012. View Article : Google Scholar : PubMed/NCBI
6	Stein P, Weber M, Prüfer S, et al: Regulatory T cells and IL-10 independently counterregulate cytotoxic T lymphocyte responses induced by transcutaneous immunization. PLoS One. 6:e279112011. View Article : Google Scholar : PubMed/NCBI
7	Mei J, Li MQ, Ding D, et al: Indoleamine 2,3-dioxygenase-1 (IDO1) enhances survival and invasiveness of endometrial stromal cells via the activation of JNK signaling pathway. Int J Clin Exp Pathol. 6:431–444. 2013.PubMed/NCBI
8	Chen SS, Corteling R, Stevanato L and Sinden J: Polyphenols inhibit indoleamine 3,5-dioxygenase-1 enzymatic activity - a role of immunomodulation in chemoprevention. Discov Med. 14:327–333. 2012.PubMed/NCBI
9	Chen S, Corteling R, Stevanato L and Sinden J: Natural inhibitors of indoleamine 3,5-dioxygenase induced by interferon-γ in human neural stem cells. Biochem Biophys Res Commun. 429:117–123. 2012. View Article : Google Scholar : PubMed/NCBI
10	Wang D, Saga Y, Mizukami H, et al: Indoleamine-2,3-dioxygenase, an immunosuppressive enzyme that inhibits natural killer cell function, as a useful target for ovarian cancer therapy. Int J Oncol. 40:929–934. 2012.
11	de Jong RA, Nijman HW, Boezen HM, et al: Serum tryptophan and kynurenine concentrations as parameters for indoleamine 2,3-dioxygenase activity in patients with endometrial, ovarian, and vulvar cancer. Int J Gynecol Cancer. 21:1320–1327. 2011.PubMed/NCBI
12	Opitz CA, Litzenburger UM, Opitz U, et al: The indoleamine-2,3-dioxygenase (IDO) inhibitor 1-methyl-D-tryptophan upregulates IDO1 in human cancer cells. PLoS One. 6:e198232011. View Article : Google Scholar : PubMed/NCBI
13	Fallarino F, Grohmann U, You S, et al: The combined effects of tryptophan starvation and tryptophan catabolites down-regulate T cell receptor ζ-chain and induce a regulatory phenotype in naive T cells. J Immunol. 176:6752–6761. 2006. View Article : Google Scholar : PubMed/NCBI
14	Erkanli S, Bolat F, Kayaselcuk F, Demirhan B and Kuscu E: COX-2 and survivin are overexpressed and positively correlated in endometrial carcinoma. Gynecol Oncol. 104:320–325. 2007. View Article : Google Scholar
15	Schwandt A, Garcia JA, Elson P, et al: Clinical and immunomodulatory effects of celecoxib plus interferon-α in metastatic renal cell carcinoma patients with COX-2 tumor immunostaining. J Clin Immunol. 31:690–698. 2011. View Article : Google Scholar : PubMed/NCBI
16	Hryniewicz A, Boasso A, Edghill-Smith Y, et al: CTLA-4 blockade decreases TGF-β, IDO, and viral RNA expression in tissues of SIVmac251-infected macaques. Blood. 108:3834–3842. 2006. View Article : Google Scholar : PubMed/NCBI
17	Wolf AM, Wolf D, Steurer M, Gastl G, Gunsilius E and Grubeck-Loebenstein B: Increase of regulatory T cells in the peripheral blood of cancer patients. Clin Cancer Res. 9:606–612. 2003.PubMed/NCBI
18	Curiel TJ, Coukos G, Zou L, et al: Specific recruitment of regulatory T cells in ovarian carcinoma fosters immune privilege and predicts reduced survival. Nat Med. 10:942–949. 2004. View Article : Google Scholar : PubMed/NCBI
19	Matsushita H, Morishita R, Nata T, et al: Hypoxia-induced endothelial apoptosis through nuclear factor-κB (NF-κB)-mediated bcl-2 suppression: in vivo evidence of the importance of NF-κB in endothelial cell regulation. Circ Res. 86:974–981. 2000. View Article : Google Scholar : PubMed/NCBI
20	He X, Brenchley PE, Jayson GC, Hampson L, Davies J and Hampson IN: Hypoxia increases heparanase-dependent tumor cell invasion, which can be inhibited by antiheparanase antibodies. Cancer Res. 64:3928–3933. 2004. View Article : Google Scholar : PubMed/NCBI
21	Munn DH and Mellor AL: Indoleamine 2,3-dioxygenase and tumor-induced tolerance. J Clin Invest. 117:1147–1154. 2007. View Article : Google Scholar : PubMed/NCBI
22	Setoguchi R, Hori S, Takahashi T and Sakaguchi S: Homeostatic maintenance of natural Foxp3⁺ CD25⁺ CD4⁺ regulatory T cells by interleukin (IL)-2 and induction of autoimmune disease by IL-2 neutralization. J Exp Med. 201:723–735. 2005. View Article : Google Scholar : PubMed/NCBI
23	Saraiva M and O’Garra A: The regulation of IL-10 production by immune cells. Nat Rev Immunol. 10:170–181. 2010. View Article : Google Scholar : PubMed/NCBI
24	Orlova VV, Liu Z, Goumans MJ and ten Dijke P: Controlling angiogenesis by two unique TGF-β type I receptor signaling pathways. Histol Histopathol. 26:1219–1230. 2011.PubMed/NCBI
25	Boniface K, Blumenschein WM, Brovont-Porth K, et al: Human Th17 cells comprise heterogeneous subsets including IFN-γ-producing cells with distinct properties from the Th1 lineage. J Immunol. 185:679–687. 2010. View Article : Google Scholar : PubMed/NCBI
26	Voskoboinik I, Dunstone MA, Baran K, Whisstock JC and Trapani JA: Perforin: structure, function, and role in human immunopathology. Immunol Rev. 235:35–54. 2010.PubMed/NCBI
27	Elpek KG, Lacelle C, Singh NP, Yolcu ES and Shirwan H: CD4⁺CD25⁺ T regulatory cells dominate multiple immune evasion mechanisms in early but not late phases of tumor development in a B cell lymphoma model. J Immunol. 178:6840–6848. 2007. View Article : Google Scholar : PubMed/NCBI
28	Lee Y and Gustafsson AB: Role of apoptosis in cardiovascular disease. Apoptosis. 14:536–548. 2009. View Article : Google Scholar : PubMed/NCBI
29	Facciabene A, Santoro S and Coukos G: Know thy enemy: Why are tumor-infiltrating regulatory T cells so deleterious? Oncoimmunology. 1:575–577. 2012. View Article : Google Scholar : PubMed/NCBI
30	Diao Z, Shi J, Zhu J, et al: TRAIL suppresses tumor growth in mice by inducing tumor-infiltrating CD4⁺CD25⁺ Treg apoptosis. Cancer Immunol Immunother. 62:653–663. 2013. View Article : Google Scholar
31	Naora H and Montell DJ: Ovarian cancer metastasis: integrating insights from disparate model organisms. Nat Rev Cancer. 5:355–366. 2005. View Article : Google Scholar : PubMed/NCBI
32	Tan DS, Agarwal R and Kaye SB: Mechanisms of transcoelomic metastasis in ovarian cancer. Lancet Oncol. 7:925–934. 2006. View Article : Google Scholar : PubMed/NCBI
33	Facciabene A, Peng X, Hagemann IS, et al: Tumour hypoxia promotes tolerance and angiogenesis via CCL28 and T(reg) cells. Nature. 475:226–230. 2011. View Article : Google Scholar : PubMed/NCBI
34	Facciabene A, Motz GT and Coukos G: T-regulatory cells: key players in tumor immune escape and angiogenesis. Cancer Res. 72:2162–2171. 2012. View Article : Google Scholar : PubMed/NCBI