[en] CONTEXT: Orexins A and B are neuropeptides that bind and activate 2 types of receptors. In addition to direct action in the brain, the orexinergic system has broader implications in peripheral organs, and it has been proposed to have a role in the induction of apoptosis. There are very few data on the endometrium. OBJECTIVE: The expression and epigenetic regulation of type 2 orexin receptor (OX2R) was investigated in the human endometrium as well as in endometrial endometrioid carcinoma (EEC). METHODS: OX2R localization was studied by immunohistochemistry in normal endometrium (n = 24) and in EEC (n = 32). The DNA methylation status of a CpG island located in the first exon of OX2R was analyzed by bisulfite sequencing in normal (n = 18), EEC (n = 34), and 3 endometrial cell lines. On the latter, mRNA expression and Western blotting as well as in vitro induction with orexin were performed. RESULTS: Expression of the OX2R protein was detected in normal endometrial epithelia, whereas it was frequently lacking in EEC. This loss was associated with hypermethylation of OX2R in EEC in comparison with normal endometrium (median CpG methylation percentages of 48.85% and 5.85%, respectively). In cell lines, hypermethylation correlated with weak OX2R expression. Additionally, in vitro treatment of the 3 EEC cell lines with orexins A and B did not result in proliferation change CONCLUSIONS: Altogether our data provide evidence for the epigenetic silencing of OX2R in EEC. The implication of the OX2R loss in tumoral progression remains to be elucidated.
Disciplines :
Oncology
Author, co-author :
Dehan, Pierre ; Université de Liège - ULiège > Département des sciences biomédicales et précliniques > Anatomie et cytologie pathologiques
Canon, C.
Trooskens, G.
Rehli, M.
Munaut, Carine ; Université de Liège - ULiège > Département des sciences cliniques > Labo de biologie des tumeurs et du développement
Van Criekinge, W.
Delvenne, Philippe ; Université de Liège - ULiège > Département des sciences biomédicales et précliniques > Anatomie et cytologie pathologiques
Language :
English
Title :
Expression of type 2 orexin receptor in human endometrium and its epigenetic silencing in endometrial cancer.
Publication date :
2013
Journal title :
Journal of Clinical Endocrinology and Metabolism
ISSN :
0021-972X
eISSN :
1945-7197
Publisher :
Oxford University Press, New York, United States - New York
de Lecea L, Kilduff TS, Peyron C, et al. The hypocretins: hypothalamus-specific peptides with neuroexcitatory activity. Proc Natl Acad Sci USA. 1998; 95: 322-327.
Sakurai T, Amemiya A, Ishii M, et al. Orexins and orexin receptors: a family of hypothalamic neuropeptides and G protein-coupled receptors that regulate feeding behavior. Cell. 1998; 92: 573-585.
Heinonen MV, Purhonen AK, Makela KA, Herzig KH. Functions of orexins in peripheral tissues. Acta Physiol (Oxf). 2008; 192: 471-485.
Kirchgessner AL, Liu M. Orexin synthesis and response in the gut. Neuron. 1999; 24: 941-951.
Mitsuma T, Hirooka Y, Kayma M, et al. Radioimmunoassay for hypocretin-2. Endocr Regul. 2000; 34: 23-27.
Mitsuma T, Hirooka Y, Kayama M, et al. Radioimmunoassay for orexin A. Life Sci. 2000; 66: 897-904.
Karteris E, Chen J, Randeva HS. Expression of human prepro-orexin and signaling characteristics of orexin receptors in the male reproductive system. J Clin Endocrinol Metab. 2004; 89: 1957-1962.
Tafuri S, Muto RL, Pavone LM, et al. Novel localization of orexin A in the tubular cytotypes of the rat testis. Regul Pept. 2010; 164: 53-57.
Silveyra P, Lux-Lantos V, Libertun C. Both orexin receptors are expressed in rat ovaries and fluctuate with the estrous cycle: effects of orexin receptor antagonists on gonadotropins and ovulation. Am J Physiol Endocrinol Metab. 2007; 293: E977-E985.
Laburthe M, Voisin T, El Firar A. Orexins/hypocretins and orexin receptors in apoptosis: a mini-review. Acta Physiol (Oxf). 2010; 198: 393-402.
Rouet-Benzineb P, Rouyer-Fessard C, Jarry A, et al. Orexins acting at native OX(1) receptor in colon cancer and neuroblastoma cells or at recombinant OX(1) receptor suppress cell growth by inducing apoptosis. J Biol Chem. 2004; 279: 45875-45886.
Nakabayashi M, Suzuki T, Takahashi K, et al. Orexin-A expression in human peripheral tissues. Mol Cell Endocrinol. 2003; 205: 43-50.
Hecht JL, Mutter GL. Molecular and pathologic aspects of endometrial carcinogenesis. J Clin Oncol. 2006; 24: 4783-4791.
You JS, Jones PA. Cancer genetics and epigenetics: two sides of the same coin? Cancer Cell. 2012; 22: 9-20.
Tao MH, Freudenheim JL. DNA methylation in endometrial cancer. Epigenetics. 2010; 5: 491-498.
Esteller M. Epigenetics in cancer. N Engl J Med. 2008; 358: 1148-1159.
Sharma S, Kelly TK, Jones PA. Epigenetics in cancer. Carcinogenesis. 2010; 31: 27-36.
Laird PW. The power and the promise of DNA methylation markers. Nat Rev Cancer. 2003; 3: 253-266.
Eckhardt F, Lewin J, Cortese R, et al. DNA methylation profiling of human chromosomes 6, 20 and 22. Nat Genet. 2006; 38: 1378-1385.
Mo B, Vendrov AE, Palomino WA, DuPont BR, Apparao KB, Lessey BA. ECC-1 cells: a well-differentiated steroid-responsive endometrial cell line with characteristics of luminal epithelium. Biol Reprod. 2006; 75: 387-394.
Hackenberg R, Loos S, Nia AH, Kunzmann R, Schulz KD. Expression of placental protein 14 by the new endometrial cancer cell line MFE-280 in vitro and by endometrial carcinomas in vivo. Anticancer Res. 1998; 18: 1153-1158.
Rohde C, Zhang Y, Reinhardt R, Jeltsch A. BISMA-fast and accurate bisulfite sequencing data analysis of individual clones from unique and repetitive sequences. BMC Bioinformatics. 2010; 11: 230.
Randeva HS, Karteris E, Grammatopoulos D, Hillhouse EW. Expression of orexin-A and functional orexin type 2 receptors in the human adult adrenals: implications for adrenal function and energy homeostasis. J Clin Endocrinol Metab. 2001; 86: 4808-4813.
Nagasaka T, Goel A, Notohara K, et al. Methylation pattern of the O6-methylguanine-DNA methyltransferase gene in colon during progressive colorectal tumorigenesis. Int J Cancer. 2008; 122: 2429-2436.
Jass JR, Whitehall VL, Young J, Leggett BA. DNA methylation in colorectal cancer. In: Szyf M, ed. DNA Methylation and Cancer Therapy. Georgetown and New York: Kluwer Academic/Plenum and Landes Biosciences/Eurekah.com Publishers; 2005: 59-68.
Rashid A, Shen L, Morris JS, Issa JP, Hamilton SR. CpG island methylation in colorectal adenomas. Am J Pathol. 2001; 159: 1129-1135.
Chen J, Randeva HS. Genomic organization and regulation of the human orexin (hypocretin) receptor 2 gene: identification of alternative promoters. Biochem J. 2010; 427: 377-390.
Brenet F, Moh M, Funk P, et al. DNA methylation of the first exon is tightly linked to transcriptional silencing. PloS One. 2011; 6: e14524.
Meaburn EL, Schalkwyk LC, Mill J. Allele-specific methylation in the human genome: implications for genetic studies of complex disease. Epigenetics. 2010; 5: 578-582.
Luedi PP, Dietrich FS, Weidman JR, Bosko JM, Jirtle RL, Hartemink AJ. Computational and experimental identification of novel human imprinted genes. Genome Res. 2007; 17: 1723-1730.
Faquin WC, Fitzgerald JT, Boynton KA, Mutter GL. Intratumoral genetic heterogeneity and progression of endometrioid type endometrial adenocarcinomas. Gynecol Oncol. 2000; 78: 152-157.
Feng YZ, Shiozawa T, Horiuchi A, et al. Intratumoral heterogeneous expression of p53 correlates with p53 mutation, Ki-67, and cyclin A expression in endometrioid-type endometrial adenocarcinomas. Virchows Archiv. 2005; 447: 816-822.
Mikeska T, Candiloro IL, Dobrovic A. The implications of heterogeneous DNA methylation for the accurate quantification of methylation. Epigenomics. 2010; 2: 561-573.
Voisin T, Firar AE, Avondo V, Laburthe M. Orexin-induced apoptosis: the key role of the seven-transmembrane domain orexin type 2 receptor. Endocrinology. 2006; 147: 4977-4984.
Spinazzi R, Rucinski M, Neri G, Malendowicz LK, Nussdorfer GG. Preproorexin and orexin receptors are expressed in cortisol-secreting adrenocortical adenomas, and orexins stimulate in vitro cortisol secretion and growth of tumor cells. J Clin Endocrinol Metab. 2005; 90: 3544-3549.
DeCarvalho DD, Sharma S, You JS, et al. DNA methylation screening identifies driver epigenetic events of cancer cell survival. Cancer Cell. 2012; 21: 655-667.
Nitkiewicz A, Smolinska N, Maleszka A, Kiezun M, Kaminski T. Localization of orexin A and orexin B in the porcine uterus. Reprod Biol. 2012; 12: 135-155.