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Quantification of chemotherapeutic target gene mRNA expression in human breast cancer biopsies: Comparison of real‐time reverse transcription‐PCR vs. Relative quantification reverse transcription‐PCR utilizing DNA sequencer analysis of PCR products by Agnes Juhasz; Paul Frankel; Catherine Cheng; Hector Rivera; Reena Vishwanath; Alice Chiu; Kim Margolin; Yun Yen; Edward M. Newman; Tim Synold; Sharon Wilczynski; Heinz‐Josef Lenz; David Gandara; Kathy S. Albain; Jeffrey Longmate; James H. Doroshow is a Medicine article available to read on EtoBox.
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## Abstract The solid tumor mRNA expression of genes related to the mechanism of action of certain antineoplastic agents is often predictive of clinical efficacy. We report here on the development of a rapid and practical real‐time RT‐PCR method to quantify genetic expression in solid tumors. The genes examined are related to the intracellular pharmacology of gemcitabine and cisplatin, two drugs that are used in the treatment of several types of advanced cancer. We evaluated target gene mRNA levels from breast tumor samples using two quantitative RT‐PCR methods: 1) an improved relative RT‐PCR method using fluorescence‐labeled primers, automated PCR set up, and GeneScan® analysis software; and 2) real‐time RT‐PCR with redesigned primers using an ABI 7900HT instrument, with additional postprocessing of the data to adjust for efficiency differences across the target genes. Using these methods, we quantified mRNA expression levels of deoxycytidine kinase (dCK), deoxycytidylate deaminase (dCDA), the M1 and M2 subunits of ribonucleotide reductase (RRM1, RRM2), and excision cross complementation group 1 (__ERCC__1) in 35 human “fresh” frozen breast cancer biopsies. While both assay method
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It is typically read by researchers, students, and practitioners in Medicine.
- Author
- Agnes Juhasz; Paul Frankel; Catherine Cheng; Hector Rivera; Reena Vishwanath; Alice Chiu; Kim Margolin; Yun Yen; Edward M. Newman; Tim Synold; Sharon Wilczynski; Heinz‐Josef Lenz; David Gandara; Kathy S. Albain; Jeffrey Longmate; James H. Doroshow
- Publisher
- John Wiley and Sons; Wiley (John Wiley & Sons); John Wiley & Sons Inc.; Wiley (ISSN 0887-8013)
- Published
- 2003
- Language
- EN
- Field
- Medicine (Health Sciences)