@Article{oncologie.2022.024373,
AUTHOR = {Xinyue Wang, Jing Xu, Qihong Gu, Dingxuan Tang, Huoyan Ji, Shaoqing Ju, Feng Wang, Lin Chen, Ruoyu Yuan},
TITLE = {A UHPLC/MS/MS Assay Based on an Isotope-Labeled Peptide for Sensitive miR-21 Detection in HCC Serum},
JOURNAL = {Oncologie},
VOLUME = {24},
YEAR = {2022},
NUMBER = {3},
PAGES = {513--526},
URL = {http://www.techscience.com/oncologie/v24n3/49723},
ISSN = {1765-2839},
ABSTRACT = {<b>Background:</b> MicroRNAs (miRNAs) have been identified as promising novel biomarkers for cancer diagnosis and
prognosis, especially for hepatocellular carcinoma (HCC). Nowadays, the expression level of miR-21 in serum
samples is a diagnostic indicator for HCC diagnosis. Thus, the quantitative determination of miRNA concentration is of significance in clinical practice. It is particularly important to establish an analytical detection method
for miR-21 in patient serum. <b>Methods:</b> The signal readout for miR-21 was based on the mass response of a reporter peptide using an isotope dilution mass spectrometry (MS) method in this work. To be more specific, miR-21
was biotinylated before being coupled with streptavidin (SA) agarose and then hybridized with a newly synthesized DNA-peptide probe. The release and purification of the sample was based on the method including trypsin
digestion, solid-phase extraction, and drying, and the detection of the reporter peptide was carried out by
UHPLC/MS/MS. The miR-21 in the corresponding samples was quantified by the ratio of the chromatographic
peak area of the redissolved polypeptide to that of the isotope-labeled polypeptide. Additionally, within the calibration range, the performance of the method (including precision, accuracy, linearity, and recovery) was evaluated. <b>Results: </b>The concentration of miR-21 was determined using the ratio of relative peak area of stable
isotope-labeled internal standard and reporter peptide, yielding a linear range of 0.1∼30.0 nM (y = 0.0818x +
0.7554, R<sup>2</sup> = 0.9586, <i>P</i> < 0.01). The limit of detection (LOD) for miR-21 was 10 pM. For y<sup>5</sup>
, the recoveries
(n = 3) were 91.36 ± 2.19%, 93.64 ± 3.55%, and 96.04 ± 2.02% for the levels of three miR-21 samples including
RL, RM, and RH, respectively. <b>Conclusions:</b> Overall, this research provides a novel analytical approach for quantitative detection of miRNAs in clinical serum samples.},
DOI = {10.32604/oncologie.2022.024373}
}