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MicroRNA as Biomarkers and Diagnostics.

ABSTRACT: MicroRNAs (miRNAs) are a group of small non-coding RNAs that are involved in regulating a range of developmental and physiological processes; their dysregulation has been associated with development of diseases including cancer. Circulating miRNAs and exosomal miRNAs have also been proposed as being useful in diagnostics as biomarkers for diseases and different types of cancer. In this review, miRNAs are discussed as biomarkers for cancer and other diseases, including viral infections, nervous system disorders, cardiovascular disorders, and diabetes. We summarize some of the clinical evidence for the use of miRNAs as biomarkers in diagnostics and provide some general perspectives on their use in clinical situations. The analytical challenges in using miRNAs in cancer and disease diagnostics are evaluated and discussed. Validation of specific miRNA signatures as biomarkers is a critical milestone in diagnostics.

SUBMITTER: Wang J

PROVIDER: S-EPMC8776330 | biostudies-literature | 2016 Jan

REPOSITORIES: biostudies-literature

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MicroRNA as Biomarkers and Diagnostics.

Wang Jin J Chen Jinyun J Sen Subrata S

Journal of cellular physiology 20160101 1

MicroRNAs (miRNAs) are a group of small non-coding RNAs that are involved in regulating a range of developmental and physiological processes; their dysregulation has been associated with development of diseases including cancer. Circulating miRNAs and exosomal miRNAs have also been proposed as being useful in diagnostics as biomarkers for diseases and different types of cancer. In this review, miRNAs are discussed as biomarkers for cancer and other diseases, including viral infections, nervous s ...[more]

PMID: 26031493

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Project description:Spermathecal fluid proteins were cleaned up via acetone precipitation overnight at 20 C. The precipitated protein was pelleted and washed twice with 500 ul of 80% acetone, then the pellet was allowed to air dry (ca. 5 min) prior to solubilization in 50 ul of digestion buffer (6 M urea, 2 M thiourea). Approximately 25 ug of protein were reduced (0.5 ug dithiothreitol, 20 min), alkylated (2.5 ug iodoacetamide, 30 min, dark), and digested (0.5 ug Lys-C for 3 h, then 0.5 ug trypsin overnight). Digested peptides were acidified with one volume of 1% trifluoroacetic acid and desalted with high-capacity STAGE tips. Eluted samples were dried (SpeedVac, Eppendorf, 45 min) and resuspended in Buffer A (0.1% formic acid). Peptide concentrations were determined using a NanoDrop (Thermo, 280 nm) and sample orders were randomized for liquid chromatography-tandem mass spectrometry (LC-MS/MS) analysis. Peptides (0.5 ug for each sample) were injected on an EASY-nLC 1000 liquid chromatography system (Thermo) coupled to an Impact II Q-TOF mass spectrometer (Bruker). The LC system included a fused-silica (5 um Aqua C18 particles (Phenomenex)) fritted 2 cm trap column connected to a 50 cm analytical column packed with ReproSil C18 (3 um C18 particles (Dr. Maisch)). The separation gradient ran from 5% to 35% Buffer B (80% acetonitrile, 0.1% formic acid) over 120 min, followed by a 15 min wash at 95% Buffer B (flow rate: 250 uL/min). The instrument parameters were: scan from 150 to 2200 m/z, 100 us transient time, 10 us prepulse storage, 7 eV collision energy, 1500 Vpp Collision RF, a +2 default charge state, 18 Hz spectral acquisition rate, 3.0 s cycle time, and the intensity threshold was 250 counts. Mass spectrometry data were searched using MaxQuant (v1.6.1.0) using default parameters, except match between runs was enabled. Peptide spectral matches, peptide identifications and protein identifications were controlled at 1% false discovery rates (FDRs). Data were analyzed in Perseus (v1.6.1.1). Proteins differentially expressed between heat-shocked and non-heat-shocked samples, as well as among tissues (semen, virgin spermathecae, and mated spermathecae), were identified using t-tests and significant results were corrected to either 5% (heat-shock and cold-shock) or 10% FDR (pesticide stress) (Banjamini-Hochberg method).

Dataset Information

MicroRNA as Biomarkers and Diagnostics.

Publications

MicroRNA as Biomarkers and Diagnostics.

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