Wei-Ting Chang1,2,3, Tzu-Ling Huang1, Zhih-Cherng Chen1,4, Yin-Hsun Feng5,6. 1. Division of Cardiology, Department of Internal Medicine, Chi-Mei Medical Center, Tainan, Taiwan. 2. Department of Biotechnology, Southern Taiwan University of Science and Technology, Tainan, Taiwan. 3. Institute of Clinical Medicine, College of Medicine, National Cheng Kung University, Tainan, Taiwan. 4. Department of Pharmacy, Chia Nan University of Pharmacy and Science, Tainan, Taiwan. 5. Division of Hematology, Department of Internal Medicine, Chi-Mei Medical Center, Tainan, Taiwan. 6. Department of Nursing, College of Medicine and Life Science, Chung-Hwa University of Medical Technology, Tainan, Taiwan.
Abstract
OBJECTIVE: Bone marrow fluid (BMF) consists of various components that establishes a microenvironment for cell differentiation and remodeling. MicroRNA-21 (miR-21) levels have recently emerged as novel biomarkers for different diseases. However, the conventional RNU6B (U6), used as the reference for intracellular miRNA, may not be appropriate for the normalization of circulating miRNAs. METHODS: We measured the levels of U6, spiked-in RNA, and miR-21 in the BMF of 13 healthy controls and 37 patients with hematological disorders to investigate the reliability of either U6 or spike-in RNA as an endogenous reference and also to study the correlation between miR-21, hematological disorders and mortality. RESULTS: Notably, the levels of U6 demonstrated a high variability in BMF of healthy controls and patients. In contrast, the levels of spiked-in RNA displayed a significantly higher stability in both cohorts. Compared with controls, the levels of miR-21 were significantly upregulated in BMF of patients with leukemia but not lymphoma. Also, using 21 as the cut-off value of miR-21, it differentiated the mortality of patients with hematologic disorders. CONCLUSIONS: Collectively, using spiked-in RNA as a reference the upregulated miR-21 levels in BMF could be an indicator of the diagnosis of leukemia and a predictor of mortality.
OBJECTIVE: Bone marrow fluid (BMF) consists of various components that establishes a microenvironment for cell differentiation and remodeling. MicroRNA-21 (miR-21) levels have recently emerged as novel biomarkers for different diseases. However, the conventional RNU6B (U6), used as the reference for intracellular miRNA, may not be appropriate for the normalization of circulating miRNAs. METHODS: We measured the levels of U6, spiked-in RNA, and miR-21 in the BMF of 13 healthy controls and 37 patients with hematological disorders to investigate the reliability of either U6 or spike-in RNA as an endogenous reference and also to study the correlation between miR-21, hematological disorders and mortality. RESULTS: Notably, the levels of U6 demonstrated a high variability in BMF of healthy controls and patients. In contrast, the levels of spiked-in RNA displayed a significantly higher stability in both cohorts. Compared with controls, the levels of miR-21 were significantly upregulated in BMF of patients with leukemia but not lymphoma. Also, using 21 as the cut-off value of miR-21, it differentiated the mortality of patients with hematologic disorders. CONCLUSIONS: Collectively, using spiked-in RNA as a reference the upregulated miR-21 levels in BMF could be an indicator of the diagnosis of leukemia and a predictor of mortality.
Entities:
Keywords:
Hematological disorders; U6; bone marrow fluid; miR-21; spiked-in RNA
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