title: A facile, branched DNA assay to quantitatively measure glucocorticoid receptor auto-regulation in T-cell acute lymphoblastic leukemia.
abstract: Glucocorticoid (GC) steroid hormones are used to treat acute lymphoblastic leukemia (ALL) because of their pro-apoptotic effects in hematopoietic cells.  However, not all leukemia cells are sensitive to GC, and no assay to stratify patients is available.  In the GC-sensitive T-cell ALL cell line CEM-C7, auto-up-regulation of RNA transcripts for the glucocorticoid receptor (GR) correlates with increased apoptotic response.  This study aimed to determine if a facile assay of GR transcript levels might be promising for stratifying ALL patients into hormone-sensitive and hormone-resistant populations.  The GR transcript profiles of various lymphoid cell lines and 4 bone marrow samples from patients with T-cell ALL were analyzed using both an optimized branched DNA (bDNA) assay and a real-time quantitative reverse transcription-polymerase chain reaction assay.  There were significant correlations between both assay platforms when measuring total GR (exon 5/6) transcripts in various cell lines and patient samples, but not for a probe set that detects a specific, low abundance GR transcript (exon 1A3).  Our results suggest that the bDNA platform is reproducible and precise when measuring total GR transcripts and, with further development, may ultimately offer a simple clinical assay to aid in the prediction of GC-sensitivity in ALL patients.
authors: Department of Biochemistry and Molecular Biology, Stanley S. Scott Cancer Center, Louisiana State University Health Science Center, New Orleans, LA 70112, USA.
authors: Jason R
authors: JR
authors: Schwartz
authors: Purvaba J
authors: PJ
authors: Sarvaiya
authors: Lily E
authors: LE
authors: Leiva
authors: Maria C
authors: MC
authors: Velez
authors: Tammuella C
authors: TC
authors: Singleton
authors: Lolie C
authors: LC
authors: Yu
authors: Wayne V
authors: WV
authors: Vedeckis
mesh_terms: D000293:Adolescent; D018931:Antineoplastic Agents, Hormonal / Q000494:pharmacology; D017209:Apoptosis / Q000187:drug effects*; D021121:Branched DNA Signal Amplification Assay / Q000379:methods*; D045744:Cell Line, Tumor; D002648:Child; D003907:Dexamethasone / Q000494:pharmacology; D019008:Drug Resistance, Neoplasm; D005091:Exons; D005938:Glucocorticoids / Q000494:pharmacology; D006801:Humans; D054218:Precursor T-Cell Lymphoblastic Leukemia-Lymphoma / Q000378:metabolism / Q000473:pathology; D011965:Receptors, Glucocorticoid / Q000235:genetics / Q000378:metabolism*; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods*; D014158:Transcription, Genetic / Q000187:drug effects; D015854:Up-Regulation
chemical_list: D018931:Antineoplastic Agents, Hormonal; D005938:Glucocorticoids; D011965:Receptors, Glucocorticoid; D003907:Dexamethasone