Importance of polymorphisms in glucocorticoid receptor and adrenocorticotropic receptor genes in development of adrenal incidentalomas

UVOD: Glukokortikoidni hormoni (GC) ostvaruju svoje efekte vezivanjem za glukokortikoidni receptor (GR). Adrenokortikotropni hormon (ACTH) reguliše sintezu GC vezivanjem za ACTH receptor (ACTHR). Prisustvo polimorfizama u genu za GR (BclI, N363S, ER22/23EK and A3669G) i promotoru ACTHR može uticati na efekte glukokortikoida i predispoziciju za nastanak unilateralnih adrenalnih incidentaloma. CILJ RADA: Utvrđivanje mogućeg uticaja funkcionalnih polimorfizama u genima za GR i ACTHR na predispoziciju za nastanak adrenalnih incidentaloma i osetljivost na GC i ispitivanje ekspresije GR u tumorskom, peritumorskom i zdravom adrenokortikalnom tkivu. METODE: U ispitivanje je bilo uključeno 112 pacijenata i 100 zdravih dobrovoljaca, koji su podvrgnuti metaboličkom, genetičkom, biohemijskom i antropometrijskom testiranju. DNK je dobijena iz leukocita periferne krvi. Prisustvo polimorfizama je detektovano metodama PCR, RFLP i sekvenciranja DNK. Uzorci tkiva su analizirani imunohistohemijskom metodom. REZULTATI: Prisustvo dužeg C alela BclI (p<0.001) polimorfizma i kraćeg G alela A3669G (p<0.001) polimorfizma GR gena su bili nezavisni prediktori adrenalnih incidentaloma. Pacijenti sa prisutnim C alelom BclI su imali veće tumore (p=0.002), a oni sa G alelom A3669G više vrednosti postdeksametazonskog kortizola (p=0.025). Istovremeno prisustvo oba alela je koreliralo sa manjim obimom struka (p=0.002), a višim baznim i postdeksametazonskog kortizolom (p=0.024). Smanjena ekspresija GRα i GRβ izoformi zapažena je u tumorskom, a GRα u peritumorskom tkivu. Lokalizacija GRβ je bila dominantno nukleusna. ZAKLJUČAK: Prisustvo C alela BclI i G alela A3669G polimorfizmama gena za GR se nalaze u vezi sa nastankom unilateralnih adrenalnim incidentalomima, a njihovo zajedničko prisustvo dovodi do smanjene osetljivosti na GC. Stečena intraadenomatozna glukokortikoidna rezistencija može da dovede do disregulacije produkcije kortizola i rasta tumora u isto vreme, dok prirodna osetljivost na glukokortikoide najverovatnije modifikuje ove efekte.INTRODUCTION: Glucocorticoid hormones (GCs) accomplish their effects through binding to glucocorticoid receptor (GR). Adrenocorticotropic hormone (ACTH) regulates synthesis of GCs through binding to ACTH receptor (ACTHR). Presence of common GR gene (BclI, N363S, ER22/23EK and A3669G) and ACTHR promoter polymorphisms can modulate GCs sensitivity. OBJECTIVE: The aim of present study was to determine weather functional polymorphisms in GR and ACTHR genes influence susceptibility for unilateral adrenal incidentalomas and GC sensitivity, and to investigate GR expression in tumorous, peritumorous and normal adrenocortical tissue samples. METHODS: The study included 112 patients with adrenal incidentalomas and 100 population-matched controls. All subjects underwent metabolic, genetic, biochemical and anthropometric testing. DNA was obtained from peripheral blood leucocytes. The polymorphisms were detected using PCR, RFLP and DNA sequencing. Tissue samples were studied by immunohistochemistry. RESULTS: GR gene variant, C allele of BclI (p<0.001) and G allele of A3669G (p<0.001) polymorphisms were independent predictors of adrenal incidentalomas. Patients with present C allele had larger tumors (p=0.002), but those with G allele had higher postdexamethasone serum cortisol (p=0.025). Both allele carriers had lesser waist circumference (p=0.002), higher basal (p=0.024) and postdexamethasone cortisol concentrations. In tumorous tissues GRα and GRβ isoforms had lower expression, but only GRα in peritumorous tissue. Localization of GRβ was dominantly nuclear. CONCLUSION: GR gene variants, larger C allele of BclI and minor 3669G allele are associated with adrenal incidentalomas. Their concurrent presence in patients reduces GC sensitivity. The acquired tumorous GC resistance probably promote dysregulated cortisol production and tumor growth, but natural sensitivity to glucocorticoides maybe modifies these effects

Oral squamous cell carcinoma detection by salivary biomarkers in a Serbian population

Early detection of oral squamous cell cancer (OSCC) is the key to improve the low 5-year survival rate. Using proteomic and genomic technologies we have previously discovered and validated salivary OSCC markers in American patients. The question arises whether these biomarkers are discriminatory in cohorts of different ethnic background. Six transcriptome (DUSP1, IL8, IL1B, OAZ1, SAT1, and S100P) and three proteome (IL1B, IL8, and M2BP) biomarkers were tested on 18 early and 17 late stage OSCC patients and 51 healthy controls with quantitative PCR and ELISA. Four transcriptome (IL8, IL1B, SAT1, and S100P) and all proteome biomarkers were significantly elevated (p lt 0.05) in OSCC patients. The combination of markers yielded an AUC of 0.86, 0.85 and 0.88 for OSCC total, T1-T2, and T3-T4, respectively. The sensitivity/specificity for OSCC total was 0.89/0.78, for T1-T2 0.67/0.96, and for T3-T4 0.82/0.84. In conclusion, seven of the nine salivary biomarkers (three proteins and four mRNAs) were validated and performed strongest in late stage cancer. Patient-based salivary diagnostics is a highly promising approach for OSCC detection. This study shows that previously discovered and validated salivary OSCC biomarkers are discriminatory and reproducible in a different ethnic cohort. These findings support the feasibility to implement multi-center, multi-ethnicity clinical trials towards the pivotal validation of salivary biomarkers for OSCC detection

Importance of polymorphisms in glucocorticoid receptor and adrenocorticotropic receptor genes in development of adrenal incidentalomas

UVOD: Glukokortikoidni hormoni (GC) ostvaruju svoje efekte vezivanjem za glukokortikoidni receptor (GR). Adrenokortikotropni hormon (ACTH) reguliše sintezu GC vezivanjem za ACTH receptor (ACTHR). Prisustvo polimorfizama u genu za GR (BclI, N363S, ER22/23EK and A3669G) i promotoru ACTHR može uticati na efekte glukokortikoida i predispoziciju za nastanak unilateralnih adrenalnih incidentaloma. CILJ RADA: Utvrđivanje mogućeg uticaja funkcionalnih polimorfizama u genima za GR i ACTHR na predispoziciju za nastanak adrenalnih incidentaloma i osetljivost na GC i ispitivanje ekspresije GR u tumorskom, peritumorskom i zdravom adrenokortikalnom tkivu. METODE: U ispitivanje je bilo uključeno 112 pacijenata i 100 zdravih dobrovoljaca, koji su podvrgnuti metaboličkom, genetičkom, biohemijskom i antropometrijskom testiranju. DNK je dobijena iz leukocita periferne krvi. Prisustvo polimorfizama je detektovano metodama PCR, RFLP i sekvenciranja DNK. Uzorci tkiva su analizirani imunohistohemijskom metodom. REZULTATI: Prisustvo dužeg C alela BclI (p<0.001) polimorfizma i kraćeg G alela A3669G (p<0.001) polimorfizma GR gena su bili nezavisni prediktori adrenalnih incidentaloma. Pacijenti sa prisutnim C alelom BclI su imali veće tumore (p=0.002), a oni sa G alelom A3669G više vrednosti postdeksametazonskog kortizola (p=0.025). Istovremeno prisustvo oba alela je koreliralo sa manjim obimom struka (p=0.002), a višim baznim i postdeksametazonskog kortizolom (p=0.024). Smanjena ekspresija GRα i GRβ izoformi zapažena je u tumorskom, a GRα u peritumorskom tkivu. Lokalizacija GRβ je bila dominantno nukleusna. ZAKLJUČAK: Prisustvo C alela BclI i G alela A3669G polimorfizmama gena za GR se nalaze u vezi sa nastankom unilateralnih adrenalnim incidentalomima, a njihovo zajedničko prisustvo dovodi do smanjene osetljivosti na GC. Stečena intraadenomatozna glukokortikoidna rezistencija može da dovede do disregulacije produkcije kortizola i rasta tumora u isto vreme, dok prirodna osetljivost na glukokortikoide najverovatnije modifikuje ove efekte.INTRODUCTION: Glucocorticoid hormones (GCs) accomplish their effects through binding to glucocorticoid receptor (GR). Adrenocorticotropic hormone (ACTH) regulates synthesis of GCs through binding to ACTH receptor (ACTHR). Presence of common GR gene (BclI, N363S, ER22/23EK and A3669G) and ACTHR promoter polymorphisms can modulate GCs sensitivity. OBJECTIVE: The aim of present study was to determine weather functional polymorphisms in GR and ACTHR genes influence susceptibility for unilateral adrenal incidentalomas and GC sensitivity, and to investigate GR expression in tumorous, peritumorous and normal adrenocortical tissue samples. METHODS: The study included 112 patients with adrenal incidentalomas and 100 population-matched controls. All subjects underwent metabolic, genetic, biochemical and anthropometric testing. DNA was obtained from peripheral blood leucocytes. The polymorphisms were detected using PCR, RFLP and DNA sequencing. Tissue samples were studied by immunohistochemistry. RESULTS: GR gene variant, C allele of BclI (p<0.001) and G allele of A3669G (p<0.001) polymorphisms were independent predictors of adrenal incidentalomas. Patients with present C allele had larger tumors (p=0.002), but those with G allele had higher postdexamethasone serum cortisol (p=0.025). Both allele carriers had lesser waist circumference (p=0.002), higher basal (p=0.024) and postdexamethasone cortisol concentrations. In tumorous tissues GRα and GRβ isoforms had lower expression, but only GRα in peritumorous tissue. Localization of GRβ was dominantly nuclear. CONCLUSION: GR gene variants, larger C allele of BclI and minor 3669G allele are associated with adrenal incidentalomas. Their concurrent presence in patients reduces GC sensitivity. The acquired tumorous GC resistance probably promote dysregulated cortisol production and tumor growth, but natural sensitivity to glucocorticoides maybe modifies these effects

Glucocorticoid Receptor and Molecular Chaperones in the Pathogenesis of Adrenal Incidentalomas: Potential Role of Reduced Sensitivity to Glucocorticoids

Glucocorticoid (GC) sensitivity depends on glucocorticoid receptor (GR) and heat shock proteins (Hsps). We investigated whether common GR genes (ER22/23EK N363S, BclI, and 9 beta) and adrenocorticotropin receptor promoter polymorphisms influence susceptibility for unilateral adrenal incidentaloma (AI), plus GR and Hsp expression in tumorous (n = 19), peritumorous (n = 13) and normal adrenocortical (n = 11) tissues. Patients (n = 112), population-matched controls (n = 100) and tumor tissues (n = 32) were genotyped for these polymorphisms. Postdexamethasone serum cortisol was higher in patients (p<0.001). GR gene variants, larger allele of BclI (odds ratio (OR) 2.9; 95% confidence interval (CI) 1.7-5.1; p < 0.001) and minor allele of 9 beta (OR 3.0; 95% CI 1.6-5.7; p < 0.001) were independent predictors of Al. In patients, the first allele is linked with larger tumors (p = 0.002) and the latter with higher postdexamethasone cortisol levels (p = 0.025). Both allele carriers had lesser waist circumference (p = 0.02), similar adrenocorticotropin and higher basal (p = 0.024) and postdexamethasone cortisol concentrations (p < 0.001). Tumorous and constitutional genotypes were similar. GR-D is the major receptor isoform in normal adrenal cortex by Western blotting. Loss of other receptor isoforms, decrease in immunostaining for GR (p < 0.0001), underexpression of chaperones (p <= 0.01) and the presence of inducible Hsp70 were found in adenomas. In conclusion, GR gene variants, C allele of BclI and minor allele of 9 beta, are associated with Als. Their concurrent presence in patients reduces GC sensitivity Normal adrenal cortex preferentially expresses GR-D. In adenomas, the lack of other GR isoforms and underexpression of heat shock proteins perhaps permanently impair GC signaling, which could promote dysregulated cortisol production and tumor growth. The innate GC sensitivity probably modifies these effects. Online address: http://www.molmed.org doi: 10.2119/molmed.2012.00261Ministry of Science and Technological Development of Serbia [III41009