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CpG island methylation status and mutation analysis of the RB1 gene essential promoter region and protein-binding pocket domain in nervous system tumours.

Gonzalez-Gomez P, Bello MJ, Alonso ME, Arjona D, Lomas J, de Campos JM, Isla A, Rey JA - Br. J. Cancer (2003)

Bottom Line: Methylation of the RB1 CpG island was detected in 26 samples corresponding to nine glioblastomas, three anaplastic astrocytomas, one mixed oligo-astrocytoma, one ependymoma, two medulloblastomas, two primary central nervous system lymphomas, two neurofibrosarcomas, and six brain metastasis from solid tumours.No inactivating mutations were found within the RB1 promoter region, whereas one glioblastoma and one oligodendroglioma displayed similar sequence variations consisting of 12 and 8 base pair deletions at intron 21.These results suggest that RB1 CpG island hypermethylation is a common epigenetic event that is associated with the development of malignant nervous system tumours.

View Article: PubMed Central - PubMed

Affiliation: Departmento de C. Experimental, Laboratorio de Oncogenetica Molecular, Hospital Universitario La Paz, Madrid, Spain.

ABSTRACT
A series of 136 nervous system tumours were studied to determine the methylation status of the CpG island contained within the promoter region of the RB1 gene, as well as mutation analysis of the essential promoter region and exons 20-24 (and surrounding intronic regions) coding for the protein-binding pocket domain. Methylation of the RB1 CpG island was detected in 26 samples corresponding to nine glioblastomas, three anaplastic astrocytomas, one mixed oligo-astrocytoma, one ependymoma, two medulloblastomas, two primary central nervous system lymphomas, two neurofibrosarcomas, and six brain metastasis from solid tumours. No inactivating mutations were found within the RB1 promoter region, whereas one glioblastoma and one oligodendroglioma displayed similar sequence variations consisting of 12 and 8 base pair deletions at intron 21. These results suggest that RB1 CpG island hypermethylation is a common epigenetic event that is associated with the development of malignant nervous system tumours.

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(A) Methylation-specific PCR of CpG island of the RB1 promoter in glioblastomas (P12 and P32) and anaplastic astrocytoma (P37). Cases P62 and P64 (glioblastomas) showed only unmethylated alleles. Positive control for methylated DNA: normal DNA from lymphocytes treated with SssI methyltransferase; normal control: DNA from a non-neoplastic brain tissue. Negative control from untreated lymphocytes DNA is not shown (L: molecular weight marker). (B) Sequence (reverse of the coding strand) analysis of bisulphite-modified DNA from tumour P12 (MR) and from non-neoplastic brain tissue (UR). Tumour DNA shows methylated cytosines (G in the reverse sequencing marked by arrows) at the represented CpG sites, whereas all CpG cytosines are unmethylated in DNA from non-neoplastic brain tissue (A in the reverse sequencing marked by arrowheads).
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fig2: (A) Methylation-specific PCR of CpG island of the RB1 promoter in glioblastomas (P12 and P32) and anaplastic astrocytoma (P37). Cases P62 and P64 (glioblastomas) showed only unmethylated alleles. Positive control for methylated DNA: normal DNA from lymphocytes treated with SssI methyltransferase; normal control: DNA from a non-neoplastic brain tissue. Negative control from untreated lymphocytes DNA is not shown (L: molecular weight marker). (B) Sequence (reverse of the coding strand) analysis of bisulphite-modified DNA from tumour P12 (MR) and from non-neoplastic brain tissue (UR). Tumour DNA shows methylated cytosines (G in the reverse sequencing marked by arrows) at the represented CpG sites, whereas all CpG cytosines are unmethylated in DNA from non-neoplastic brain tissue (A in the reverse sequencing marked by arrowheads).

Mentions: Results from Bello et al, (1994) and unpublished data.


CpG island methylation status and mutation analysis of the RB1 gene essential promoter region and protein-binding pocket domain in nervous system tumours.

Gonzalez-Gomez P, Bello MJ, Alonso ME, Arjona D, Lomas J, de Campos JM, Isla A, Rey JA - Br. J. Cancer (2003)

(A) Methylation-specific PCR of CpG island of the RB1 promoter in glioblastomas (P12 and P32) and anaplastic astrocytoma (P37). Cases P62 and P64 (glioblastomas) showed only unmethylated alleles. Positive control for methylated DNA: normal DNA from lymphocytes treated with SssI methyltransferase; normal control: DNA from a non-neoplastic brain tissue. Negative control from untreated lymphocytes DNA is not shown (L: molecular weight marker). (B) Sequence (reverse of the coding strand) analysis of bisulphite-modified DNA from tumour P12 (MR) and from non-neoplastic brain tissue (UR). Tumour DNA shows methylated cytosines (G in the reverse sequencing marked by arrows) at the represented CpG sites, whereas all CpG cytosines are unmethylated in DNA from non-neoplastic brain tissue (A in the reverse sequencing marked by arrowheads).
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Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC2376780&req=5

fig2: (A) Methylation-specific PCR of CpG island of the RB1 promoter in glioblastomas (P12 and P32) and anaplastic astrocytoma (P37). Cases P62 and P64 (glioblastomas) showed only unmethylated alleles. Positive control for methylated DNA: normal DNA from lymphocytes treated with SssI methyltransferase; normal control: DNA from a non-neoplastic brain tissue. Negative control from untreated lymphocytes DNA is not shown (L: molecular weight marker). (B) Sequence (reverse of the coding strand) analysis of bisulphite-modified DNA from tumour P12 (MR) and from non-neoplastic brain tissue (UR). Tumour DNA shows methylated cytosines (G in the reverse sequencing marked by arrows) at the represented CpG sites, whereas all CpG cytosines are unmethylated in DNA from non-neoplastic brain tissue (A in the reverse sequencing marked by arrowheads).
Mentions: Results from Bello et al, (1994) and unpublished data.

Bottom Line: Methylation of the RB1 CpG island was detected in 26 samples corresponding to nine glioblastomas, three anaplastic astrocytomas, one mixed oligo-astrocytoma, one ependymoma, two medulloblastomas, two primary central nervous system lymphomas, two neurofibrosarcomas, and six brain metastasis from solid tumours.No inactivating mutations were found within the RB1 promoter region, whereas one glioblastoma and one oligodendroglioma displayed similar sequence variations consisting of 12 and 8 base pair deletions at intron 21.These results suggest that RB1 CpG island hypermethylation is a common epigenetic event that is associated with the development of malignant nervous system tumours.

View Article: PubMed Central - PubMed

Affiliation: Departmento de C. Experimental, Laboratorio de Oncogenetica Molecular, Hospital Universitario La Paz, Madrid, Spain.

ABSTRACT
A series of 136 nervous system tumours were studied to determine the methylation status of the CpG island contained within the promoter region of the RB1 gene, as well as mutation analysis of the essential promoter region and exons 20-24 (and surrounding intronic regions) coding for the protein-binding pocket domain. Methylation of the RB1 CpG island was detected in 26 samples corresponding to nine glioblastomas, three anaplastic astrocytomas, one mixed oligo-astrocytoma, one ependymoma, two medulloblastomas, two primary central nervous system lymphomas, two neurofibrosarcomas, and six brain metastasis from solid tumours. No inactivating mutations were found within the RB1 promoter region, whereas one glioblastoma and one oligodendroglioma displayed similar sequence variations consisting of 12 and 8 base pair deletions at intron 21. These results suggest that RB1 CpG island hypermethylation is a common epigenetic event that is associated with the development of malignant nervous system tumours.

Show MeSH
Related in: MedlinePlus