PMID: 7519188Jan 1, 1994Paper

Competitive nested polymerase chain reaction for quantification of human MDR1 gene expression

Journal of Cancer Research and Clinical Oncology
F GrünebachK Schumacher

Abstract

Tumor cell resistance to cytotoxic drugs is considered one of the major obstacles to successful chemotherapy. Multidrug resistance (MDR) describes the simultaneous expression of cellular resistance to a wide range of structurally and functionally unrelated drugs. The development of the multidrug resistance phenotype is accompanied by multiple morphological and biochemical changes: (a) increased glutathione levels in the cytoplasm, (b) modified levels of enzymes in the nucleus, particularly topoisomerase II, (c) increased DNA repair capacity and (d) overexpression of the (human) MDR1 gene encoding a transmembrane efflux pump (P-glycoprotein, gp-170), which leads to decreased intracellular accumulation and therefore to resistance to a variety of cytotoxic drugs. In this report we describe a competitive polymerase chain reaction (PCR) assay for the absolute quantification of MDR1 mRNA. This assay uses a transcript generated in vitro as an internal standard which is later coamplified together with the MDR1 cDNA. Both cDNAs exhibit the same MDR1 primer sites but differ in the length of the amplicon. For a second round of amplification we applied nested MDR1 primers and were successful in improving the sensitivity of this competitive...Continue Reading

References

Sep 1, 1990·Proceedings of the National Academy of Sciences of the United States of America·K E NoonanI B Roninson
Jan 1, 1987·Proceedings of the National Academy of Sciences of the United States of America·A T FojoI Pastan
Dec 1, 1989·Proceedings of the National Academy of Sciences of the United States of America·A M WangD F Mark
Jan 18, 1989·Journal of the National Cancer Institute·L J GoldsteinG M Brodeur
Apr 1, 1989·Journal of Clinical Oncology : Official Journal of the American Society of Clinical Oncology·W S DaltonS E Salmon

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Citations

Jul 24, 1998·Clinical Chemistry and Laboratory Medicine : CCLM·C OrlandoM Pazzagli
May 21, 2013·Proceedings of the National Academy of Sciences of the United States of America·Sabine KussJanine Mauzeroll

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