Seladin-1 expression is regulated by promoter methylation in adrenal cancer.
BACKGROUND
Seladin-1 overexpression exerts a protective mechanism against apoptosis. Seladin-1 mRNA is variably expressed in normal human tissues. Adrenal glands show the highest levels of seladin-1 expression, which are significantly reduced in adrenal carcinomas (ACC). Since up to now seladin-1 mutations were not described, we investigated whether promoter methylation could account for the down-regulation of seladin-1 expression in ACC.


METHODS
A methylation sensitive site was identified in the seladin-1 gene. We treated DNA extracted from two ACC cell lines (H295R and SW13) with the demethylating agent 5-Aza-2-deoxycytidine (5-Aza). Furthermore, to evaluate the presence of an epigenetic regulation also 'in vivo', seladin-1 methylation and its mRNA expression were measured in 9 ACC and in 5 normal adrenal glands.


RESULTS
The treatment of cell lines with 5-Aza induced a significant increase of seladin-1 mRNA expression in H295R (fold increase, F.I. = 1.8; p = 0.02) and SW13 (F.I. = 2.9; p = 0.03). In ACC, methylation density of seladin-1 promoter was higher (2682 +/- 686) than in normal adrenal glands (362 +/- 97; p = 0.02). Seladin-1 mRNA expression in ACC (1452 +/- 196) was significantly lower than in normal adrenal glands (3614 +/- 949; p = 0.01).


CONCLUSIONS
On this basis, methylation could be involved in the altered pattern of seladin-1 gene expression in ACC.
Simi
Lisa
L
Clinical Biochemistry, Department of Clinical Physiopathology, University of Florence, viale Pieraccini 6, Florence, 50139 Italy.
Malentacchi
Francesca
F
Luciani
Paola
P
Gelmini
Stefania
S
Deledda
Cristiana
C
Arvia
Rosaria
R
Mannelli
Massimo
M
Peri
Alessandro
A
Orlando
Claudio
C
D000310:Adrenal Gland Neoplasms / Q000201:enzymology / Q000235:genetics*; D001374:Azacitidine / Q000031:analogs & derivatives / Q000494:pharmacology; D002277:Carcinoma / Q000201:enzymology / Q000235:genetics*; D045744:Cell Line, Tumor; D019175:DNA Methylation / Q000187:drug effects; D015254:DNA Modification Methylases / Q000037:antagonists & inhibitors / Q000378:metabolism; D000077209:Decitabine; D004791:Enzyme Inhibitors / Q000494:pharmacology; D044127:Epigenesis, Genetic / Q000187:drug effects; D015971:Gene Expression Regulation, Enzymologic; D015972:Gene Expression Regulation, Neoplastic; D006801:Humans; D009419:Nerve Tissue Proteins / Q000235:genetics* / Q000378:metabolism; D044925:Oxidoreductases Acting on CH-CH Group Donors / Q000235:genetics* / Q000378:metabolism; D016133:Polymerase Chain Reaction; D011401:Promoter Regions, Genetic / Q000187:drug effects; D012333:RNA, Messenger / Q000378:metabolism; D013997:Time Factors
D004791:Enzyme Inhibitors; D009419:Nerve Tissue Proteins; D012333:RNA, Messenger; D000077209:Decitabine; D044925:Oxidoreductases Acting on CH-CH Group Donors; C499055:DHCR24 protein, human; D015254:DNA Modification Methylases; D001374:Azacitidine
Genes Dev. 1995 Jan 1;9(1):97-107
7828854
Mol Cell Biol. 2008 Jan;28(2):539-50
17984220
J Clin Endocrinol Metab. 2002 Mar;87(3):1170-6
11889182
FASEB J. 2002 Oct;16(12):1526-36
12374775
J Cell Biochem. 2004 Jan 1;91(1):54-69
14689582
Int J Cancer. 2005 Jun 10;115(2):224-30
15688385
Clin Endocrinol (Oxf). 2005 Jul;63(1):111-5
15963070
Cancer Res. 2006 May 15;66(10):5012-20
16707422
Proc Natl Acad Sci U S A. 2001 May 8;98(10):5856-61
11296263
Neurobiol Dis. 2008 Mar;29(3):422-37
18086530
J Clin Endocrinol Metab. 2004 Mar;89(3):1332-9
15001630
Eur Urol. 2006 Aug;50(2):290-301
16455181
J Endocrinol. 2007 Jan;192(1):53-66
17210742
Endocrinology. 2008 Jul;149(7):3267-73
18339707
Nature. 2004 Dec 2;432(7017):640-5
15577914
J Clin Endocrinol Metab. 2001 Nov;86(11):5130-7
11701665
J Neurosci. 2000 Oct 1;20(19):7345-52
11007892
Ann N Y Acad Sci. 2002 Dec;975:232-46
12538168
Neuroscience. 2002;113(2):301-10
12127087
Clin Biochem. 2003 Feb;36(1):61-5
12554062
Cancer Genomics Proteomics. 2005 Apr;2(2):97-114
18548127
J Clin Endocrinol Metab. 2005 Nov;90(11):6156-61
16091489
J Mol Endocrinol. 2004 Dec;33(3):651-62
15591025
Int J Oncol. 2003 Nov;23(5):1425-30
14532986
EMBO J. 2006 Jan 25;25(2):432-43
16407971
Clin Cancer Res. 2004 Nov 15;10(22):7457-65
15569975
J Neurochem. 2007 Nov;103 Suppl 1:159-70
17986151