التفاصيل البيبلوغرافية
| العنوان: |
Concurrent epigenetic silencing of wnt/β-catenin pathway inhibitor genes in B cell chronic lymphocytic leukaemia |
| المؤلفون: |
Moskalev, Evgeny A, Luckert, Katrin, Vorobjev, Ivan A, Mastitsky, Sergey E, Gladkikh, Aleena A, Stephan, Achim, Schrenk, Marita, Kaplanov, Kamil D, Kalashnikova, Olga B, Pötz, Oliver, Joos, Thomas O, Hoheisel, Jörg D |
| بيانات النشر: |
BioMed Central Ltd. |
| سنة النشر: |
2012 |
| المجموعة: |
BioMed Central |
| مصطلحات موضوعية: |
B cell chronic lymphocytic leukaemia, Wnt/β-catenin pathway, Inhibitor genes, DNA hypermethylation, Epigenetic silencing, β-catenin |
| الوصف: |
Background The Wnt/β-catenin signalling is aberrantly activated in primary B cell chronic lymphocytic leukaemia (CLL). Epigenetic silencing of pathway inhibitor genes may be a mechanism for its activation. In this study, we investigated systematically and quantitatively the methylation status of 12 Wnt/β-catenin pathway inhibitor genes – CDH1, DACT1, DKK1, DKK2, DKK3, DKK4, SFRP1, SFRP2, SFRP3, SFRP4, SFRP5 and WIF1 – in the cell lines EHEB and MEC-1 as well as patient samples. Methods Quantification of DNA methylation was performed by means of bisulphite pyrosequencing and confirmed by bisulphite Sanger sequencing. Gene expression was analysed by qPCR using GAPDH as internal control. E-cadherin and β-catenin protein quantification was carried out by microsphere-based immunoassays. Methylation differences observed between the patient and control groups were tested using generalised least squares models. Results For 10 genes, a higher methylation level was observed in tumour material. Only DKK4 exhibited similarly high methylation levels in both tumour and normal specimens, while DACT1 was always essentially unmethylated. However, also for these inhibitors, treatment of cells with the demethylating agent 5-aza-2´-deoxycytidine resulted in an induction of their expression, as shown by quantitative PCR, suggesting an indirect epigenetic control of activity. While the degree of demethylation and its transcriptional consequences differed between the genes, there was an overall high correlation of demethylation and increased activity. Protein expression studies revealed that no constitutive Wnt/β-catenin signalling occurred in the cell lines, which is in discrepancy with results from primary CLL. However, treatment with 5-aza-2´-deoxycytidine caused accumulation of β-catenin. Simultaneously, E-cadherin expression was strongly induced, leading to the formation of a complex with β-catenin and thus demonstrating its epigenetically regulated inhibition effect. Conclusions The results suggest an epigenetic ... |
| نوع الوثيقة: |
article in journal/newspaper |
| اللغة: |
English |
| العلاقة: |
http://www.biomedcentral.com/1471-2407/12/213Test |
| الإتاحة: |
http://www.biomedcentral.com/1471-2407/12/213Test |
| حقوق: |
Copyright 2012 Moskalev et al.; licensee BioMed Central Ltd. |
| رقم الانضمام: |
edsbas.26DDD37F |
| قاعدة البيانات: |
BASE |
