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Detection of CpG island hypermethylation of caspase-8 in neuroblastoma using an oligonucleotide array.
Pediatr Blood Cancer. 2009 Jul; 52(7):777-83.PB

Abstract

BACKGROUND

The caspase-8 gene (CASP8) is frequently inactivated in unfavorable neuroblastomas through DNA methylation. The present study utilized oligoarrays to evaluate the methylation status of a CpG island located between exons 2 and 3 of caspase 8 in neuroblastomas.

PROCEDURE

DNA derived from 70 neuroblastomas was amplified by PCR after bisulfate modification and subjected to analysis on a self-made oligoarray that utilized a polycarbodiimide-coated slide to detect methylation of six intragenic CpG islands of caspase 8. In 30 cases, the methylation status was also analyzed by sequencing. In six cases, the PCR product was cloned into a vector and analyzed.

RESULTS

Among the 70 tumor-derived DNAs, methylation was not detected in 18 cases, one methylated CpG was found in 12 cases, two in 18 cases, three in 3 cases, four in 8 cases, five in 1 case and six in 10 cases. All methylated CpG loci detected by sequencing were detected by oligoarray, but some methylated CpGs in three loci were detected by oligoarray alone. In these discrepant loci, methylation was detected in some clones after subcloning, indicating that the oligoarray might be more sensitive than sequencing. The CASP8 expression level was depressed in the tumors having two distinct CpG doublets. These results were significantly correlated with MYCN amplification and with clinical outcomes.

CONCLUSIONS

A significant difference in the methylation status within the CpG island of CASP8 was shown between favorable and unfavorable subtypes, and CASP8 methylation detected by oligoarray may be useful in the clinical evaluation of neuroblastomas.

Authors+Show Affiliations

Department of Surgery, Graduate School of Biomedical Science, Hiroshima University, Hiroshima, Japan.No affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info available

Pub Type(s)

Journal Article

Language

eng

PubMed ID

19260109

Citation

Kamimatsuse, Arata, et al. "Detection of CpG Island Hypermethylation of Caspase-8 in Neuroblastoma Using an Oligonucleotide Array." Pediatric Blood & Cancer, vol. 52, no. 7, 2009, pp. 777-83.
Kamimatsuse A, Matsuura K, Moriya S, et al. Detection of CpG island hypermethylation of caspase-8 in neuroblastoma using an oligonucleotide array. Pediatr Blood Cancer. 2009;52(7):777-83.
Kamimatsuse, A., Matsuura, K., Moriya, S., Fukuba, I., Yamaoka, H., Fukuda, E., Kamei, N., Hiyama, K., Sueda, T., & Hiyama, E. (2009). Detection of CpG island hypermethylation of caspase-8 in neuroblastoma using an oligonucleotide array. Pediatric Blood & Cancer, 52(7), 777-83. https://doi.org/10.1002/pbc.21977
Kamimatsuse A, et al. Detection of CpG Island Hypermethylation of Caspase-8 in Neuroblastoma Using an Oligonucleotide Array. Pediatr Blood Cancer. 2009;52(7):777-83. PubMed PMID: 19260109.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Detection of CpG island hypermethylation of caspase-8 in neuroblastoma using an oligonucleotide array. AU - Kamimatsuse,Arata, AU - Matsuura,Kaoru, AU - Moriya,Shogo, AU - Fukuba,Ikuko, AU - Yamaoka,Hiroaki, AU - Fukuda,Emi, AU - Kamei,Naomi, AU - Hiyama,Keiko, AU - Sueda,Taijiro, AU - Hiyama,Eiso, PY - 2009/3/5/entrez PY - 2009/3/5/pubmed PY - 2009/5/6/medline SP - 777 EP - 83 JF - Pediatric blood & cancer JO - Pediatr Blood Cancer VL - 52 IS - 7 N2 - BACKGROUND: The caspase-8 gene (CASP8) is frequently inactivated in unfavorable neuroblastomas through DNA methylation. The present study utilized oligoarrays to evaluate the methylation status of a CpG island located between exons 2 and 3 of caspase 8 in neuroblastomas. PROCEDURE: DNA derived from 70 neuroblastomas was amplified by PCR after bisulfate modification and subjected to analysis on a self-made oligoarray that utilized a polycarbodiimide-coated slide to detect methylation of six intragenic CpG islands of caspase 8. In 30 cases, the methylation status was also analyzed by sequencing. In six cases, the PCR product was cloned into a vector and analyzed. RESULTS: Among the 70 tumor-derived DNAs, methylation was not detected in 18 cases, one methylated CpG was found in 12 cases, two in 18 cases, three in 3 cases, four in 8 cases, five in 1 case and six in 10 cases. All methylated CpG loci detected by sequencing were detected by oligoarray, but some methylated CpGs in three loci were detected by oligoarray alone. In these discrepant loci, methylation was detected in some clones after subcloning, indicating that the oligoarray might be more sensitive than sequencing. The CASP8 expression level was depressed in the tumors having two distinct CpG doublets. These results were significantly correlated with MYCN amplification and with clinical outcomes. CONCLUSIONS: A significant difference in the methylation status within the CpG island of CASP8 was shown between favorable and unfavorable subtypes, and CASP8 methylation detected by oligoarray may be useful in the clinical evaluation of neuroblastomas. SN - 1545-5017 UR - https://www.unboundmedicine.com/medline/citation/19260109/Detection_of_CpG_island_hypermethylation_of_caspase_8_in_neuroblastoma_using_an_oligonucleotide_array_ L2 - https://doi.org/10.1002/pbc.21977 DB - PRIME DP - Unbound Medicine ER -