72 research outputs found
Altered SMRT levels disrupt vitamin D3 receptor signalling in prostate cancer cells.
We hypothesized that key antiproliferative target genes for the vitamin D receptor (VDR) were repressed by an epigenetic mechanism in prostate cancer cells resulting in apparent hormonal insensitivity. To explore this possibility, we examined nuclear receptor corepressor expression in a panel of nonmalignant and malignant cell lines and primary cultures, and found frequently elevated SMRT corepressor mRNA expression often associated with reduced sensitivity to 1alpha,25-dihydroxyvitamin D(3) (1alpha,25(OH)2D3). For example, PC-3 and DU-145 prostate cancer cell lines had 1.8-fold and twofold increases in SMRT mRNA relative to normal PrEC cells (P<0.05). Similarly, 10/15 primary tumour cultures (including three matched to normal cells from the same donors) had elevated SMRT mRNA levels; generally NCoR1 and Alien were not as commonly elevated. Corepressor proteins often have associated histone deacetylases (HDAC) and reflectively the antiproliferative action of 1alpha,25(OH)2D3 can be restored by cotreatment with low doses of HDAC inhibitors such as trichostatin A (TSA, 15 nM) to induce apoptosis in prostate cancer cell lines. To decipher the transcriptional events that lead to these cellular responses, we undertook gene expression studies in PC-3 cells after cotreatment of 1alpha,25(OH)2D3 plus TSA after 6 h. Examination of known VDR target genes and cDNA microarray analyses revealed cotreatment of 1alpha,25(OH)2D3 plus TSA cooperatively upregulated eight (out of 1176) genes, including MAPK-APK2 and GADD45alpha. MRNA and protein time courses and inhibitor studies confirmed these patterns of regulation. Subsequently, we knocked down SMRT levels in PC-3 cells using a small interfering RNA (siRNA) approach and found that GADD45alpha induction by 1alpha,25(OH)2D3 alone became very significantly enhanced. The same distortion of gene responsiveness, with repressed induction of GADD45alpha was found in primary tumour cultures compared and to matched peripheral zone (normal) cultures from the same donor. These data demonstrate that elevated SMRT levels are common in prostate cancer cells, resulting in suppression of target genes associated with antiproliferative action and apparent 1alpha,25(OH)2D3-insensitivity. This can be targeted therapeutically by combination treatments with HDAC inhibitors
Π Π²ΠΎΠΏΡΠΎΡΡ ΠΏΠΎΠ΄Π³ΠΎΡΠΎΠ²ΠΊΠΈ ΠΌΠ΅Π΄ΠΈΡΠΈΠ½ΡΠΊΠΈΡ ΠΊΠ°Π΄ΡΠΎΠ² Π² ΡΡΠ»ΠΎΠ²ΠΈΡΡ ΡΠ΅ΡΠΎΡΠΌΠΈΡΠΎΠ²Π°Π½ΠΈΡ ΡΠΈΡΡΠ΅ΠΌΡ Π·Π΄ΡΠ°Π²ΠΎΠΎΡ ΡΠ°Π½Π΅Π½ΠΈΡ
Π Π΅ΡΠΎΡΠΌΠΈΡΠΎΠ²Π°Π½ΠΈΠ΅ ΡΠΈΡΡΠ΅ΠΌΡ Π·Π΄ΡΠ°Π²ΠΎΠΎΡ
ΡΠ°Π½Π΅Π½ΠΈΡ Π² Π£ΠΊΡΠ°ΠΈΠ½Π΅ Π²Π½ΠΎΡΠΈΡ ΡΠ΅ΡΡΠ΅Π·Π½ΡΠ΅ ΠΊΠΎΡΒΡΠ΅ΠΊΡΠΈΠ²Ρ Π² ΡΠ°Π±ΠΎΡΡ Π»Π΅ΡΠ΅Π±Π½ΠΎ-ΠΏΡΠΎΡΠΈΠ»Π°ΠΊΡΠΈΡΠ΅ΡΠΊΠΈΡ
ΡΡΡΠ΅ΠΆΠ΄Π΅Π½ΠΈΠΉ, ΠΈ ΠΎΠΏΡΠ΅Π΄Π΅Π»ΡΠ΅Ρ Π½Π΅ΠΎΠ±Ρ
ΠΎΠ΄ΠΈΠΌΠΎΡΡΡ ΠΏΠΎΠ΄Π³ΠΎΡΠΎΠ²ΠΊΠΈ, ΠΏΠ΅ΡΠ΅ΠΏΠΎΠ΄Π³ΠΎΡΠΎΠ²ΠΊΠΈ, ΠΏΠΎΠ²ΡΡΠ΅Π½ΠΈΡ ΠΊΠ²Π°Π»ΠΈΡΠΈΠΊΠ°ΡΠΈΠΈ, ΠΊΠ°ΠΊ Π²ΡΠ°ΡΠ΅ΠΉ ΡΠ°ΠΊ ΠΈ
ΡΡΠ΅Π΄Π½Π΅Π³ΠΎ ΠΌΠ΅Π΄ΠΈΡΠΈΠ½ΡΠΊΠΎΠ³ΠΎ ΠΏΠ΅ΡΡΠΎΠ½Π°Π»Π°. Π‘ΠΈΡΡΠ΅ΠΌΠ° Π΄ΠΎΠΏΠΎΠ»Π½ΠΈΡΠ΅Π»ΡΠ½ΠΎΠ³ΠΎ ΠΏΡΠΎΡΠ΅ΡΡΠΈΠΎΠ½Π°Π»ΡΠ½ΠΎΠ³ΠΎ ΠΎΠ±ΡΠ°Π·ΠΎΠ²Π°Π½ΠΈΡ Π΄ΠΎΠ»ΠΆΠ½Π° Π±ΡΡΡ ΠΎΠ΄Π½ΠΎΠΉ ΠΈΠ· ΠΎΡΠ½ΠΎΠ²ΠΎΠΏΠΎΠ»Π°Π³Π°ΡΡΠΈΡ
ΡΠΈΡΡΠ΅ΠΌ, ΠΎΠ±Π΅ΡΠΏΠ΅ΡΠΈΠ²Π°ΡΡΠΈΡ
ΠΏΡΠ°ΠΊΡΠΈΡΠ΅ΡΠΊΠΎΠ΅ Π·Π΄ΡΠ°Π²ΠΎΠΎΡ
ΡΠ°Π½Π΅Π½ΠΈΠ΅ Π²ΡΡΠΎΠΊΠΎΠΏΡΠΎΡΠ΅ΡΡΠΈΠΎΠ½Π°Π»ΡΠ½ΡΠΌΠΈ ΠΊΠ°Π΄ΡΠ°ΠΌΠΈ. Π Π΅Π°Π»ΠΈΠ·Π°ΡΠΈΡ
ΡΡΠΎΠ³ΠΎ Π²ΠΎΠ·ΠΌΠΎΠΆΠ½Π° ΠΏΡΠΈ ΠΈΡΠΏΠΎΠ»ΡΠ·ΠΎΠ²Π°Π½ΠΈΠΈ ΡΠΎΠ²ΡΠ΅ΠΌΠ΅Π½Π½ΡΡ
, Π°ΠΊΡΡΠ°Π»ΡΠ½ΡΡ
ΠΈ Π°ΠΏΡΠΎΠ±ΠΈΡΠΎΠ²Π°Π½Π½ΡΡ
ΠΌΠ΅ΡΠΎΠ΄ΠΈΠΊ ΠΎΠ±ΡΡΠ΅Π½ΠΈΡ, ΠΏΠ΅ΡΠ΅ΠΎΠ±ΡΡΠ΅Π½ΠΈΡ ΠΈ ΠΏΠΎΠ²ΡΡΠ΅Π½ΠΈΡ ΠΊΠ²Π°Π»ΠΈΡΠΈΠΊΠ°ΡΠΈΠΈ
HumanMethylation450K arrayβidentified biomarkers predict tumour recurrence/progression at initial diagnosis of high-risk non-muscle Invasive bladder cancer
Background: High-risk non-muscle invasive bladder cancer (HR-NMIBC) is a clinically unpredictable disease. Despite clinical risk estimation tools, many patients are undertreated with intra-vesical therapies alone, whereas others may be over-treated with early radical surgery.
Molecular biomarkers, particularly DNA methylation, have been reported as predictive of tumour/patient outcomes in numerous solid organ and haematologic malignancies; however, there are few reports in HR-NMIBC and none using genome-wide array assessment. We therefore sought to identify novel DNA methylation markers of HR-NMIBC clinical outcomes that might predict tumour behaviour at initial diagnosis and help guide patient management.
Patients and methods: A total of 21 primary initial diagnosis HR-NMIBC tumours were analysed by Illumina HumanMethylation450 BeadChip arrays and subsequently bisulphite Pyrosequencing. In all, 7 had not recurred at 1 year after resection and 14 had recurred and/or progressed despite intra-vesical BCG. A further independent cohort of 32 HR-NMIBC tumours (17 no recurrence and 15 recurrence and/ or progression despite BCG) were also assessed by bisulphite Pyrosequencing.
Results: Array analyses identified 206 CpG loci that segregated non-recurrent HR-NMIBC tumours from clinically more aggressive recurrence/progression tumours. Hypermethylation of CpG cg11850659 and hypomethylation of CpG cg01149192 in combination predicted HRNMIBC recurrence and/or progression within 1 year of diagnosis with 83% sensitivity, 79% specificity, and 83% positive and 79% negative predictive values.
Conclusions: This is the first genome-wide DNA methylation analysis of a unique HR-NMIBC tumour cohort encompassing known 1-year clinical outcomes. Our analyses identified potential novel epigenetic markers that could help guide individual patient management in this clinically unpredictable diseas
Π‘ΡΡΠ°ΡΠ½ΠΈΠΉ ΠΏΠΎΠ³Π»ΡΠ΄ Π½Π° ΠΏΡΠΎΠ±Π»Π΅ΠΌΡ Π±ΡΠΎΠ½Ρ ΠΎΠ»Π΅Π³Π΅Π½Π΅Π²ΠΎΡ Π΄ΠΈΡΠΏΠ»Π°Π·ΡΡ ΡΠ° Π²ΡΠ΄ΠΊΡΠΈΡΠΎΡ Π°ΡΡΠ΅ΡΡΠ°Π»ΡΠ½ΠΎΡ ΠΏΡΠΎΡΠΎΠΊΠΈ Ρ Π½Π΅Π΄ΠΎΠ½ΠΎΡΠ΅Π½ΠΈΡ Π½ΠΎΠ²ΠΎΠ½Π°ΡΠΎΠ΄ΠΆΠ΅Π½ΠΈΡ
Π£ ΡΡΠ°ΡΡΡ Π²ΡΠ΄ΠΎΠ±ΡΠ°ΠΆΠ΅Π½Ρ ΡΡΡΠ°ΡΠ½Ρ Π°ΡΠΏΠ΅ΠΊΡΠΈ ΠΏΠ°ΡΠΎΠ³Π΅Π½Π΅Π·Ρ ΡΠ° ΠΊΠ»ΡΠ½ΡΠΊΠΈ Π±ΡΠΎΠ½Ρ
ΠΎΠ»Π΅Π³Π΅Π½Π΅Π²ΠΎΡ Π΄ΠΈΡΠΏΠ»Π°Π·ΡΡ Ρ Π½Π΅Π΄ΠΎΠ½ΠΎΡΠ΅Π½ΠΈΡ
Π½ΠΎΠ²ΠΎΠ½Π°ΡΠΎΠ΄ΠΆΠ΅Π½ΠΈΡ
. ΠΠ°Π΄Π°Π½ΠΎ Π°Π½Π°Π»ΡΠ· Π²ΠΏΠ»ΠΈΠ²Ρ ΡΡΠ·Π½ΠΈΡ
Π°Π½ΡΠ΅Π½Π°ΡΠ°Π»ΡΠ½ΠΈΡ
ΡΠ° ΡΠ½ΡΡΠ°Π½Π°ΡΠ°Π»ΡΠ½ΠΈΡ
ΡΠ°ΠΊΡΠΎΡΡΠ² Π½Π° ΡΠΎΡΠΌΡΠ²Π°Π½Π½Ρ ΡΠ° ΠΏΠ΅ΡΠ΅Π±ΡΠ³ Π±ΡΠΎΠ½Ρ
ΠΎΠ»Π΅Π³Π΅Π½Π΅Π²ΠΎΡ Π΄ΠΈΡΠΏΠ»Π°Π·ΡΡ. ΠΠΊΡΠ΅ΠΌΠΎ ΡΠΎΠ·Π³Π»ΡΠ½ΡΡΠΎ ΠΏΡΠΎΠ±Π»Π΅ΠΌΡ Π²ΡΠ΄ΠΊΡΠΈΡΠΎΡ Π°ΡΡΠ΅ΡΡΠ°Π»ΡΠ½ΠΎΡ ΠΏΡΠΎΡΠΎΠΊΠΈ,
ΠΏΠ°ΡΠΎΠ³Π΅Π½Π΅Π·, ΠΌΠ΅Ρ
Π°Π½ΡΠ·ΠΌΠΈ Π²ΠΏΠ»ΠΈΠ²Ρ Π½Π° ΡΠΎΠ·Π²ΠΈΡΠΎΠΊ Π±ΡΠΎΠ½Ρ
ΠΎΠ»Π΅Π³Π΅Π½Π΅Π²ΠΎΡ Π΄ΠΈΡΠΏΠ»Π°Π·ΡΡ ΡΠ° ΠΎΠ±ΡΡΠΆΠ΅Π½Π½Ρ ΡΡ ΡΡΠ°Π½Ρ. Π ΠΎΠ·Π³Π»ΡΠ½ΡΡΠΎ ΠΊΠ»ΡΠ½ΡΡΠ½Ρ
ΡΠ° ΠΏΠ°ΡΠΎΠ³Π΅Π½Π΅ΡΠΈΡΠ½Ρ Π°ΠΏΠ΅ΠΊΡΠΈ ΡΡΠ½ΠΊΡΡΠΎΠ½ΡΠ²Π°Π½Π½Ρ Π²ΡΠ΄ΠΊΡΠΈΡΠΎΡ Π°ΡΡΠ΅ΡΡΠ°Π»ΡΠ½ΠΎΡ ΠΏΡΠΎΡΠΎΠΊΠΈ Ρ Π½Π΅Π΄ΠΎΠ½ΠΎΡΠ΅Π½ΠΈΡ
Π½ΠΎΠ²ΠΎΠ½Π°ΡΠΎΠ΄ΠΆΠ΅Π½ΠΈΡ
.
. The article reflects the modern aspects
of the pathogenesis and clinic of bronchopulmonary
dysplasia in premature infants. An analysis of the
impact of various antenatal and intranatal factors on
the development and course of bronchopulmonary
dysplasia. Separately the problem of patent ductus
arteriosus. Pathogenesis, mechanisms of influence on
the development of bronchopulmonary dysplasia and
encumbrance of her condition. Discussed the clinical
and pathogenetic apects functioning of patent ductus
arteriosus in preterm infants
Nuclear Receptors and the Warburg effect in cancer
In 1927 Otto Warburg established that tumours derive energy primarily from the conversion of glucose to lactic acid and only partially through cellular respiration involving oxygen. In the 1950βs he proposed that all causes of cancer reflected different mechanisms of disabling cellular respiration in favour of fermentation (now termed aerobic glycolysis). The role of aberrant glucose metabolism in cancer is now firmly established. The shift away from oxidative phosphorylation towards the metabolically expensive aerobic glycolysis is somewhat counter-intuitive given its wasteful nature. Multiple control processes are in place to maintain cellular efficiency and it is likely that these mechanisms are disrupted to facilitate the shift to the reliance on aerobic glycolysis. One such process of cell control is mediated by the nuclear receptor superfamily. This large family of transcription factors plays a significant role in sensing environmental cues and controlling decisions on proliferation, differentiation and cell death for example, to regulate glucose uptake and metabolism and to modulate the actions of oncogenes and tumour suppressors. In this review we highlight mechanisms by which nuclear receptors actions are altered during tumorigenic transformation and can serve to enhance the shift to aerobic glycolysis. At the simplest level, a basic alteration in NR behaviour can serve to enhance glycolytic flux thus providing a basis for enhanced survival within the tumour micro-environment. Ameliorating the enhanced NR activity in this context may help to sensitize cancer cells to Warburg targeted therapies and may provide future drug targets
Quantitative genome-wide methylation analysis of high-grade non-muscle invasive bladder cancer
High-grade non-muscle invasive bladder cancer (HG-NMIBC) is a clinically unpredictable disease with greater risks of recurrence and progression relative to their low-intermediate-grade counterparts. The molecular events, including those affecting the epigenome, that characterise this disease entity in the context of tumour development, recurrence and progression, are incompletely understood. We therefore interrogated genome-wide DNA methylation using HumanMethylation450 BeadChip-arrays in 21 primary HG-NMIBC tumours relative to normal bladder controls. Using strict inclusion-exclusion criteria we identified 1,057 hypermethylated CpGs within gene promoter-associated CpG islands, representing 256 genes. Bisulphite Pyrosequencing validated the array data and examined 25 array-identified candidate genes in an independent cohort of 30 HG-NMIBC and 18 low-intermediate-grade NMIBC. These analyses revealed significantly higher methylation frequencies in high-grade tumours relative to low-intermediate-grade tumours for the ATP5G2, IRX1 and VAX2 genes (p<0.05), and similarly significant increases in mean levels of methylation in high-grade tumours for the ATP5G2, VAX2, INSRR, PRDM14, VSX1, TFAP2b, PRRX1, and HIST1H4F genes (p<0.05). Although inappropriate promoter methylation was not invariantly associated with reduced transcript expression, a significant association was apparent for the ARHGEF4, PON3, STAT5a, and VAX2 gene transcripts (p<0.05). Herein, we present the first genome-wide DNA methylation analysis in a unique HG-NMIBC cohort, showing extensive and discrete methylation changes relative to normal bladder and low-intermediate-grade tumours. The genes we identified hold significant potential as targets for novel therapeutic intervention either alone, or in combination, with more conventional therapeutic options in the treatment of this clinically unpredictable disease
HumanMethylation450K arrayβidentified biomarkers predict tumour recurrence/progression at initial diagnosis of high-risk non-muscle Invasive bladder cancer
Background: High-risk non-muscle invasive bladder cancer (HR-NMIBC) is a clinically unpredictable disease. Despite clinical risk estimation tools, many patients are undertreated with intra-vesical therapies alone, whereas others may be over-treated with early radical surgery.Molecular biomarkers, particularly DNA methylation, have been reported as predictive of tumour/patient outcomes in numerous solid organ and haematologic malignancies; however, there are few reports in HR-NMIBC and none using genome-wide array assessment. We therefore sought to identify novel DNA methylation markers of HR-NMIBC clinical outcomes that might predict tumour behaviour at initial diagnosis and help guide patient management.Patients and methods: A total of 21 primary initial diagnosis HR-NMIBC tumours were analysed by Illumina HumanMethylation450 BeadChip arrays and subsequently bisulphite Pyrosequencing. In all, 7 had not recurred at 1 year after resection and 14 had recurred and/or progressed despite intra-vesical BCG. A further independent cohort of 32 HR-NMIBC tumours (17 no recurrence and 15 recurrence and/ or progression despite BCG) were also assessed by bisulphite Pyrosequencing.Results: Array analyses identified 206 CpG loci that segregated non-recurrent HR-NMIBC tumours from clinically more aggressive recurrence/progression tumours. Hypermethylation of CpG cg11850659 and hypomethylation of CpG cg01149192 in combination predicted HRNMIBC recurrence and/or progression within 1 year of diagnosis with 83% sensitivity, 79% specificity, and 83% positive and 79% negative predictive values.Conclusions: This is the first genome-wide DNA methylation analysis of a unique HR-NMIBC tumour cohort encompassing known 1-year clinical outcomes. Our analyses identified potential novel epigenetic markers that could help guide individual patient management in this clinically unpredictable diseas
Methylation of HOXA9 and ISL1 predicts patient outcome in high-grade non-invasive bladder cancer
Introduction
Inappropriate DNA methylation is frequently associated with human tumour development, and in specific cases, is associated with clinical outcomes. Previous reports of DNA methylation in low/intermediate grade non-muscle invasive bladder cancer (NMIBC) have suggested that specific patterns of DNA methylation may have a role as diagnostic or prognostic biomarkers. In view of the aggressive and clinically unpredictable nature of high-grade (HG) NMIBC, and the current shortage of the preferred treatment option (Bacillus:Calmette-Guerin), novel methylation analyses may similarly reveal biomarkers of disease outcome that could risk-stratify patients and guide clinical management at initial diagnosis.
Methods
Promoter-associated CpG island methylation was determined in primary tumour tissue of 36 initial presentation high-grade NMIBCs, 12 low/intermediate-grade NMIBCs and 3 normal bladder controls. The genes HOXA9, ISL1, NKX6-2, SPAG6, ZIC1 and ZNF154 were selected for investigation on the basis of previous reports and/or prognostic utility in low/intermediate-grade NMIBC. Methylation was determined by Pyrosequencing of sodium-bisulphite converted DNA, and then correlated with gene expression using RT-qPCR. Methylation was additionally correlated with tumour behaviour, including tumour recurrence and progression to muscle invasive bladder cancer or metastases.
Results
The ISL1 genesβ promoter-associated island was more frequently methylated in recurrent and progressive high-grade tumours than their non-recurrent counterparts (60.0% vs. 18.2%, p = 0.008). ISL1 and HOXA9 showed significantly higher mean methylation in recurrent and progressive tumours compared to non-recurrent tumours (43.3% vs. 20.9%, p = 0.016 and 34.5% vs 17.6%, p = 0.017, respectively). Concurrent ISL1/HOXA9 methylation in HG-NMIBC reliably predicted tumour recurrence and progression within one year (Positive Predictive Value 91.7%), and was associated with disease-specific mortality (DSM).
Conclusions
In this study we report methylation differences and similarities between clinical sub-types of high-grade NMIBC. We report the potential ability of methylation biomarkers, at initial diagnosis, to predict tumour recurrence and progression within one year of diagnosis. We found that specific biomarkers reliably predict disease outcome and therefore may help guide patient treatment despite the unpredictable clinical course and heterogeneity of high-grade NMIBC. Further investigation is required, including validation in a larger patient cohort, to confirm the clinical utility of methylation biomarkers in high-grade NMIBC
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