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Promoter methylatio...
Promoter methylation in the PTCH gene in cervical epithelial cancer and ovarian cancer tissue as studied by eight novel Pyrosequencing® assays
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Löf-Öhlin, Zarah M (författare)
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Levanat, Sonja (författare)
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Sabol, Maja (författare)
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visa fler...
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Sorbe, Bengt (författare)
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Nilsson, Torbjörn K (författare)
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visa färre...
- 2011
- 2011
- Engelska.
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Ingår i: International Journal of Oncology. - 1019-6439. ; 38:3, 685-692
Abstract
Ämnesord
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- DNA methylation status in the CpG sites of promoter regions in cancer-related genes, such as PTCH, has traditionally been investigated using either dye-terminator sequencing or methylation-specific PCR. We aimed to study the PTCH gene promoter methylation in gynecological cancers, with a method that gives a quantitative measure of the methylation status of the promoter region of the studied gene, and for this purpose, we designed novel Pyrosequencing-based assays. Bisulfite-treated genomic DNA (bsDNA) was amplified by standard PCR and applied to novel Pyrosequencing® assays, in order to measure the methylated fraction (%) at each CpG site of the PTCH gene promoter. We analyzed 22 squamous cell cervical cancer tissue specimens (11 with good and 11 with poor outcomes after radiotherapy) and 5 ovarian cancer tissue specimens matched with 5 normal ovarian tissue specimens. Six optimized PCR protocols which generated 8 Pyrosequencing assays covering 63 CpG sites in the promoter regions 1 and 2 as well as the previously unanalyzed promoter region 3 in the PTCH gene were developed. The 27 tumor tissue specimens and 5 normal tissues did not show any methylation within any of the 63 CpG sites. Our data suggest that methylation of the PTCH promoter is not a high-prevalence feature of squamous cell cervical cancer or ovarian cancer, but Pyrosequencing assays are a good method for studying promoter methylation.
Ämnesord
- Medical and Health Sciences (hsv)
- Clinical Medicine (hsv)
- Cancer and Oncology (hsv)
- Medicin och hälsovetenskap (hsv)
- Klinisk medicin (hsv)
- Cancer och onkologi (hsv)
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