Exploring allelic imbalance within paraffin-embedded tumor biopsies using pyrosequencing technology
| Autor: | Reposo Ramírez-Lorca, Manuel Hidalgo Pascual, Jose Luis Royo, María Lopez-García, Francisco Jose Martínez-Tello, Eduardo Ferrero Herrero, Agustín Ruiz, Ana Salinas, Carmen Crespo, Luis Miguel Real |
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| Rok vydání: | 2006 |
| Předmět: |
Heterozygote
Biopsy Clinical Biochemistry Single-nucleotide polymorphism Allelic Imbalance Protein Serine-Threonine Kinases Biology Polymorphism Single Nucleotide Sensitivity and Specificity Loss of heterozygosity Automation Breast cancer Neoplasms medicine Humans Allele Gene Thyroid cancer Paraffin Embedding Base Sequence Chromosomes Human Pair 11 Biochemistry (medical) Chromosome Mapping Nuclear Proteins Reproducibility of Results Sequence Analysis DNA General Medicine medicine.disease Molecular biology Neoplasm Proteins Repressor Proteins p21-Activated Kinases Pyrosequencing |
| Zdroj: | Clinical Chemistry and Laboratory Medicine (CCLM). 44 |
| ISSN: | 1437-4331 1434-6621 |
| Popis: | Background: The comparison of molecular genetic changes in healthy and pathological tissues has historically led to the identification of oncogenes and tumor suppressor genes. It is very common that studies investigating loss of heterozygosity are carried out retrospectively on paraffin-embedded samples. Methods: In this study, we evaluated the power of pyrosequencing for determining the loss of heterozygotic regions. The present method uses the fact that pyrosequencing is an accurate, sensitive and reproducible technique. The method is also simple to perform, with results available in 96-well format, making the assays amenable to automation. Thus, we analyzed nine single nucleotide polymorphisms along 1 Mb between the EMSY and PAK1 genes on 11q13, a region frequently rearranged in different tumors and cell lines. We assessed the study using samples from breast cancer and thyroid cancer biopsies. Results and conclusions: We conclude that this technique is capable of detecting variations of >10% in allele loss. However, strong allele imbalances were detected, depending on the origin of the sample. Seven out of the nine markers used exhibited differential allele amplification, depending on the DNA quality (p |
| Databáze: | OpenAIRE |
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