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Promoter methylation in the PTCH gene in cervical epithelial cancer and ovarian cancer tissue as studied by eight novel Pyrosequencing (R) assays
Örebro University, School of Health and Medical Sciences.ORCID iD: 0000-0002-5063-631X
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2011 (English)In: International Journal of Oncology, ISSN 1019-6439, Vol. 38, no 3, p. 685-692Article in journal (Refereed) Published
Abstract [en]

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(R) 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.

Place, publisher, year, edition, pages
2011. Vol. 38, no 3, p. 685-692
Keywords [en]
CpG sites, DNA methylation, Hedgehog pathway, Pyrosequencing technology
National Category
Medical and Health Sciences
Research subject
Medicine
Identifiers
URN: urn:nbn:se:oru:diva-17115DOI: 10.3892/ijo.2011.895ISI: 000287829200010PubMedID: 21206974Scopus ID: 2-s2.0-79952010237OAI: oai:DiVA.org:oru-17115DiVA, id: diva2:438813
Available from: 2011-09-05 Created: 2011-09-02 Last updated: 2017-12-08Bibliographically approved

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Sorbe, BengtNilsson, Torbjörn K.

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CiteExportLink to record
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Citation style
  • apa
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