AI Article Synopsis

  • The study introduces a nonradioactive method for quantifying specific mRNA levels through competition between labeled and unlabeled DNA sequences during hybridization.
  • The technique demonstrated changes in glucocorticoid receptor (GR) expression in U937 cells after applying differentiation agents, TPA and a RA/VD combination.
  • Results indicated TPA treatment led to increased GR mRNA levels and specific dexamethasone binding, while RA/VD treatment did not show enhanced GR expression.

Article Abstract

We describe a method for relative quantification of specific mRNA using a nonradioactive assay based on DNA strand competition between identical sequences of biotin- and fluorescein-labeled amplicon (probe) and unlabeled amplicon (target) during hybridization. As the target quantity increased, that of the double-labeled probe decreased in accordance with the mass action law. This technique was successfully applied to evaluate differences in glucocorticoid receptor expression in U937 cells before and after the addition of potent differentiation inducers: 12-O-tetradecanoylphorbol 13-acetate (TPA) and a combination of all-trans retinoic acid (RA) and 1,25-dihydroxyvitamin D2 (VD). We observed that TPA treatment was associated with an increase in specific binding of [3H]dexamethasone and up-regulation of GR mRNA while no enhanced GR expression was perceived with RA/VD treatment.

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http://dx.doi.org/10.1006/abio.1995.1275DOI Listing

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