A Method to Monitor mRNA Levels in Human Breast Tumor Cells Obtained by Fine-needle Aspiration
| Autor: | Bert Top, Sjoerd Rodenhuis, Wolter J. Mooi, Caro Lambrechts, R. A. Maas, Laura J. vanʼt Veer, Hans Peterse, Majella S. de Lange |
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| Rok vydání: | 1997 |
| Předmět: |
Porphobilinogen deaminase
Gene Expression Breast Neoplasms Biology Polymerase Chain Reaction Pathology and Forensic Medicine Biopsy Gene expression Tumor Cells Cultured medicine Humans RNA Messenger Molecular Biology Gene Messenger RNA medicine.diagnostic_test BRCA1 Protein Biopsy Needle Reproducibility of Results Cancer Cell Biology Blotting Northern medicine.disease Molecular biology Fine-needle aspiration Cell culture Feasibility Studies Regression Analysis Female |
| Zdroj: | Diagnostic Molecular Pathology. 6:353-360 |
| ISSN: | 1052-9551 |
| DOI: | 10.1097/00019606-199712000-00008 |
| Popis: | A method based on the reverse transcriptase-polymerase chain reaction (RT-PCR) was developed that allows the determination of relative mRNA expression levels in fine-needle aspirates from human tumors. The method was developed for the c-erbB-2 gene, using the porphobilinogen deaminase (PBGD) gene as an internal standard. It was validated for mRNA isolated from cell lines and for material obtained by fine-needle aspiration from human breast cancer. Gene expression levels were determined by measuring the activity of radiolabeled RT-PCR-amplified gene-specific bands with a phosphor imager. At least four points are measured on the log-linear part of the amplification cycle versus signal intensity curves, and subsequently the distance between the curves of the gene of interest and that of an internal standard gene is used to calculate the relative expression levels. The method worked equally well with the BRCA1 gene, illustrating that it can be generalized to other genes. The method is suitable to measure or monitor semiquantitively gene expression levels in accessible human tumors in situ. |
| Databáze: | OpenAIRE |
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