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Rebellato, Andrea (2018) Methylation status of Vitamin D receptor gene promoter in adrenocortical carcinoma. [Ph.D. thesis]

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Abstract (english)

Context: Previous data of my research group showed a decreased expression of vitamin D receptor (VDR) mRNA/protein in a small group of adrenocortical carcinoma (ACC) tissues, suggesting the loss of a protective role of VDR against malignant cell growth in this type of cancer. Downregulation of VDR gene expression may result from epigenetic events, that is, methylation of cytosine nucleotide of CpG islands in VDR gene promoter.
Objective: To analyse methylation of CpG sites in the VDR gene promoter in a series of ACC tissue specimens, comparing malignant adrenocortical tumour samples with those from various benign forms and normal adrenals.
Methods: Methylation of CpG-rich 5′ regions was assessed by bisulfite sequencing PCR using bisulfite-treated DNA from archival microdissected paraffin-embedded adrenocortical tissues. Three normal adrenals and twenty-three various adrenocortical tumour samples including eight carcinomas and fifteen adenomas were studied.
Results: Methylation in the promoter region of VDR gene was found in three of eight ACCs, while no VDR gene methylation was observed in normal adrenals and adrenocortical adenomas. VDR mRNA and protein levels were lower in ACCs than in benign tumours. VDR immunostaining was weak or negative in ACCs, including all three methylated tissue samples.
Conclusion: The association between VDR gene promoter methylation and reduced VDR gene expression is not a rare event in ACC, suggesting that VDR epigenetic inactivation may have a role in adrenocortical carcinogenesis. Other epigenetic mechanisms in the upstream signalling pathway involved in silencing VDR gene expression in ACC should be investigated.

Abstract (italian)

Contesto: Dati precedenti del gruppo di ricerca nel quale ho svolto l’attività di dottorato avevano indicato una ridotta espressione di mRNA e proteina del recettore della vitamina D (VDR) in un piccolo gruppo di campioni di carcinoma corticosurrenalico (ACC), suggerendo la perdita di un ruolo protettivo di VDR nei confronti della crescita cellulare maligna in questo tipo di cancro. La ridotta espressione del gene VDR potrebbe essere il risultato di eventi epigenetici, tra cui la metilazione dei nucleotidi di citosina delle isole CpG contenute nel promotore del gene VDR.
Oggetto: Analisi della metilazione dei siti CpG nel promotore del gene VDR in una serie di campioni di tessuto di ACC, confrontando campioni di tumori corticosurrenalici maligni e benigni e di corteccia surrenalica normale.
Metodi: La metilazione delle regioni ricche di CpG situate al 5’ è stata valutata con sequenziamento mediante PCR del DNA, estratto da campioni di corticosurrene microsezionato inclusi in paraffina e trattato con bisolfito. Sono stati studiati ventitré campioni di tumori corticosurrenalici, di cui otto carcinomi e quindici adenomi, oltre a tre campioni di corteccia surrenalica normale.
Risultati: La metilazione nella regione del promotore del gene VDR è stata documentata in tre degli otto campioni di ACC, mentre non è stata riscontrata metilazione a carico del gene VDR nei campioni di adenoma e nei campioni di tessuto di corticosurrene normale. I livelli di mRNA e proteina di VDR sono risultati minori negli ACC rispetto ai tumori benigni. L’espressione di VDR valutata con tecnica immunoistochimica è risultata debole o negativa nei campioni di ACC, inclusi i tre campioni risultati metilati.
Conclusione: L’associazione tra metilazione del promotore nel gene VDR e ridotta espressione del gene VDR non è un evento raro nell’ACC, suggerendo che l’inattivazione epigenetica di VDR potrebbe rivestire un ruolo nel processo di carcinogenesi corticosurrenalica. Meritano di essere indagati altri meccanismi epigenetici nella via di segnale a monte coinvolta nel silenziamento dell’espressione del gene VDR.

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EPrint type:Ph.D. thesis
Tutor:Fallo, Francesco
Ph.D. course:Ciclo 29 > Corsi 29 > SCIENZE MEDICHE, CLINICHE E SPERIMENTALI
Data di deposito della tesi:30 January 2018
Anno di Pubblicazione:30 January 2018
Key Words:Carcinoma corticosurrenalico, Vitamina D, VDR, Metilazione, Epigenetica / Adrenocortical carcinoma, Vitamin D, VDR, Methylation, Epigenetics
Settori scientifico-disciplinari MIUR:Area 06 - Scienze mediche > MED/13 Endocrinologia
Struttura di riferimento:Dipartimenti > Dipartimento di Medicina
Codice ID:11097
Depositato il:15 Nov 2018 12:28
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