A facile, branched DNA assay to quantitatively measure glucocorticoid receptor auto-regulation in T-cell acute lymphoblastic leukemia

• Published in: Ai zheng 2012-08, Vol.31 (8), p.381-391
• Main Authors: Schwartz, Jason R, Sarvaiya, Purvaba J, Leiva, Lily E, Velez, Maria C, Singleton, Tammuella C, Yu, Lolie C, Vedeckis, Wayne V
• Format: Article
• Language: English
• Subjects: Adolescent; Antineoplastic Agents, Hormonal - pharmacology; Apoptosis - drug effects; Branched DNA Signal Amplification Assay - methods; Cell Line, Tumor; Child; Dexamethasone - pharmacology; DNA检测; Drug Resistance, Neoplasm; Exons; Glucocorticoids - pharmacology; Humans; Original; Precursor T-Cell Lymphoblastic Leukemia-Lymphoma - metabolism; Precursor T-Cell Lymphoblastic Leukemia-Lymphoma - pathology; Receptors, Glucocorticoid - genetics; Receptors, Glucocorticoid - metabolism; Reverse Transcriptase Polymerase Chain Reaction - methods; Transcription, Genetic - drug effects; T细胞; Up-Regulation; 定量测量; 急性; 支链; 淋巴细胞白血病; 糖皮质激素受体; 自动调节
• ISSN: 1000-467X; 1944-446X
• DOI: 10.5732/cjc.012.10044

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.