Methylation-independent silencing of the p73 gene in neuroblastoma

Banelli, Barbara; Casciano, Ida; Romani, Massimo

doi:10.1038/sj.onc.1203807

Cited by 31 publications

(28 citation statements)

References 21 publications

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“…107 Silencing of the active maternal allele by methylation has been observed in acute leukaemia and Burkitt's lymphoma 108 but it is possible that methylationindependent means of silencing occur in other cancers. 109 p57KIP2, a cyclin dependent kinase inhibitor, is expressed primarily from the maternal allele. 110 In lung cancers with 11p15 deletions, the maternal allele is predominantly deleted.…”

Section: Dysregulation Of Genomic Imprinting In Cancermentioning

confidence: 99%

DNA methylation, imprinting and cancer

Plass¹,

Soloway

2002

Eur J Hum Genet

139

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It is well known that a variety of genetic changes influence the development and progression of cancer. These changes may result from inherited or spontaneous mutations that are not corrected by repair mechanisms prior to DNA replication. It is increasingly clear that so called epigenetic effects that do not affect the primary sequence of the genome also play an important role in tumorigenesis. This was supported initially by observations that cancer genomes undergo changes in their methylation state and that control of parental allele-specific methylation and expression of imprinted loci is lost in several cancers. Many loci acquiring aberrant methylation in cancers have since been identified and shown to be silenced by DNA methylation. In many cases, this mechanism of silencing inactivates tumour suppressors as effectively as frank mutation and is one of the cancer-predisposing hits described in Knudson's two hit hypothesis. In contrast to mutations which are essentially irreversible, methylation changes are reversible, raising the possibility of developing therapeutics based on restoring the normal methylation state to cancer-associated genes. Development of such therapeutics will require identifying loci undergoing methylation changes in cancer, understanding how their methylation influences tumorigenesis and identifying the mechanisms regulating the methylation state of the genome. The purpose of this review is to summarise what is known about these issues.

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Section: Dysregulation Of Genomic Imprinting In Cancermentioning

confidence: 99%

DNA methylation, imprinting and cancer

Plass¹,

Soloway

2002

Eur J Hum Genet

139

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“…In fact we did not observe a different level of methylation in primary NB expressing or not expressing TAp73. 10 In an attempt to understand the mechanisms regulating DNp73 expression we have analyzed the methylation status of the DNp73 5' region in neuroblastoma and control cell lines by Methylation Specific PCR (MSP). This technique relies upon the selective PCR amplification of methylated or unmethylated sequences after chemical conversion of the unmethylated C into T. The presence of an amplification band only with the primer set designed to bind to the methylated sequence indicated that the DNp73 5' end is fully methylated in the DNp73-negative cell lines, U937, HL60 and ACN.…”

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confidence: 99%

“…A human DNA sequence with 78% homology to nucleotides 1 ± 274 of the mouse DNp73 was identified by blast query against the human genome sequence. The human exon 3' and flanking sequences were subcloned from mini-libraries constructed from PAC clones 863N7 and 967O8 as described 10 utilizing a probe derived from the mouse cDNA sequence. The left part of the panel reports the schematic representation of the structure of the p73 locus; P1 indicates the TA promoter and P2 is the putative DNp73 promoter.…”

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confidence: 99%

Role of methylation in the control of ΔNp73 expression in neuroblastoma

et al. 2002

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“…To determine the concordance between MSP and Meth-DOP-PCR, we first assessed the methylation status of six genes (TA-p73, DN-p73, CDKN2A, RASSF1A, MGMT and DCR2) by MSP and by Meth-DOP-PCR in cell lines where the methylation status of these target genes was previously demonstrated in independent experiments (Banelli et al, 19,21 Di Vinci et al 20 and our unpublished observations). As shown in Figure 4, the six genes considered in this analysis presented an identical methylation status after MSP or Meth-DOP-PCR indicating that the results obtained by the two techniques are qualitatively concordant.…”

Section: Accuracy Of Meth-dop-pcrmentioning

confidence: 99%

“…In these cells the TA-p73 promoter is completely methylated and unmethylated, respectively. 19 Briefly, serial dilutions of DNA ranging from 500 to 7.8 ng ( Figure 2) were modified with sodium bisulfite in separate reactions and dissolved in 30 ml. Two microliter aliquots of the modified DNA that, assuming no loss of material, correspond to 33-0.51 ng of modified DNA, were subjected to Meth-DOP-PCR in a final volume of 50 ml.…”

Section: Overview and Design Of The Techniquementioning

confidence: 99%

Meth-DOP-PCR: an assay for the methylation profiling of trace amounts of DNA extracted from bodily fluids

Vinci

Gelvi

Banelli

et al. 2006

Laboratory Investigation

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Cancer cells release their DNA into the patient's bodily fluids and cancer-specific signatures can be recognized in the circulating DNA. The aberrant methylation of CpG-rich regions in gene promoter sequences is an early marker of cell transformation whose specificity and optimal sensitivity can be achieved by assessing the methylation status of multiple genes ('methylation profiling'). Most of the current technologies for methylation analysis rely upon the combination of chemical conversion of the DNA and PCR analysis for the detection of methylated and unmethylated alleles. However, the small amount of circulating DNA, and its fragmentation, dramatically reduces the template DNA molecules making difficult the methylation profiling. To overcome this limitation, we have developed the Meth-DOP-PCR assay, a combination between a modified degenerate oligonucleotide primed PCR (DOP-PCR) and methylation-specific PCR (MSP), for the high-throughput methylation analysis of trace-amount of circulating DNA. We have demonstrated the concordance between Meth-DOP-PCR and MSP and shown the application of this technique for the methylation analysis of DNA extracted from the serum of lung cancer patients. We have estimated that through this procedure it is possible to obtain at least a 25-fold increase of the number of determinations allowing the methylation profiling from less than 1 ml of serum. Thus, Meth-DOP-PCR appears as a simple, cost-effective and efficient technique, for the development of novel methylation-based diagnostic assays.

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Methylation-independent silencing of the p73 gene in neuroblastoma

Cited by 31 publications

References 21 publications

DNA methylation, imprinting and cancer

DNA methylation, imprinting and cancer

Role of methylation in the control of ΔNp73 expression in neuroblastoma

Meth-DOP-PCR: an assay for the methylation profiling of trace amounts of DNA extracted from bodily fluids

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