{"title":"Differential circulating miRNA profiles identified miR-423-5p, miR-93-5p, and miR-4532 as potential biomarkers for cholangiocarcinoma diagnosis.","authors":"Kittiya Supradit, Sattrachai Prasopdee, Teva Phanaksri, Sithichoke Tangphatsornruang, Montinee Pholhelm, Siraphatsorn Yusuk, Kritiya Butthongkomvong, Kanokpan Wongprasert, Jutharat Kulsantiwong, Amnat Chukan, Smarn Tesana, Veerachai Thitapakorn","doi":"10.7717/peerj.18367","DOIUrl":null,"url":null,"abstract":"<p><strong>Background: </strong>Cholangiocarcinoma (CCA) is high in morbidity and mortality rates which may be due to asymptomatic and effective diagnostic methods not available. Therefore, an effective diagnosis is urgently needed.</p><p><strong>Methods: </strong>Investigation of plasma circulating miRNA (cir-miRNA) was divided into two phases, including the discovery phase (pooled 10 samples each from three pools in each group) and the validation phase (17, 16, and 35 subjects of healthy control (HC), <i>O. viverrini</i> (OV), and CCA groups, respectively). The plasma from healthy control subjects, <i>O. viverrini</i> infected subjects, and CCA subjects was used. In the discovery phase, plasma was pooled by adding an equal volume of plasma, and cir-miRNA was isolated and analyzed with the nCounter<sup>®</sup> SPRINT Profiler. The significantly different cir-miRNAs were selected for the validation phase. In the validation phase, cir-miRNA was isolated and analyzed using real time-quantitative polymerase chain reaction (RT-qPCR). Subsequently, statistical analysis was conducted, and diagnostic parameters were calculated.</p><p><strong>Results: </strong>Differential plasma cir-miRNA profile showed at least three candidates including miR-423-5p, miR-93-5p, and miR-4532 as potential biomarkers. From validation of these cir-miRNAs by RT-qPCR, the result showed that the satisfied sensitivity and specificity to differential CCA group from HC and OV group was obtained from miR-4532 (<i>P</i> < 0.05) while miR-423-5p and miR-93-5p can be used for differential CCA from OV and HC group (<i>P</i> < 0.05) with high specificity but limited the sensitivity. In conclusion, candidate cir-miRNAs have been identified as potential biomarkers including miR-423-5p, miR-93-5p and miR-4532. Screening by miR-4532 and confirmed with miR-423-5p, miR-93-5p were suggested for differential CCA patients in the endemic area of <i>O. viverrini</i>.</p>","PeriodicalId":19799,"journal":{"name":"PeerJ","volume":"12 ","pages":"e18367"},"PeriodicalIF":2.3000,"publicationDate":"2024-12-10","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11639864/pdf/","citationCount":"0","resultStr":null,"platform":"Semanticscholar","paperid":null,"PeriodicalName":"PeerJ","FirstCategoryId":"99","ListUrlMain":"https://doi.org/10.7717/peerj.18367","RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"2024/1/1 0:00:00","PubModel":"eCollection","JCR":"Q2","JCRName":"MULTIDISCIPLINARY SCIENCES","Score":null,"Total":0}
引用次数: 0
摘要
背景:胆管癌(Colangiocarcinoma,CCA)的发病率和死亡率都很高,这可能是由于无症状和缺乏有效的诊断方法所致。因此,迫切需要有效的诊断方法:方法:血浆循环 miRNA(cir-miRNA)的研究分为两个阶段,包括发现阶段(每组 3 个样本池各 10 个样本)和验证阶段(健康对照组(HC)、O. viverrini 组(OV)和 CCA 组分别有 17、16 和 35 名受试者)。使用的血浆分别来自健康对照组、O. viverrini 感染组和 CCA 组。在发现阶段,加入等体积的血浆进行汇集,用 nCounter® SPRINT Profiler 分离和分析 cir-miRNA。筛选出差异明显的 cir-miRNA 用于验证阶段。在验证阶段,使用实时定量聚合酶链反应(RT-qPCR)分离和分析 cir-miRNA。随后进行统计分析,计算诊断参数:结果:血浆cir-miRNA差异图谱显示,至少有三个候选的潜在生物标志物,包括miR-423-5p、miR-93-5p和miR-4532。通过RT-qPCR对这些cir-miRNAs进行验证,结果表明,miR-4532对鉴别HC组和OV组CCA的敏感性和特异性都很满意(P<0.05),而miR-423-5p和miR-93-5p可用于鉴别OV组和HC组CCA(P<0.05),特异性高,但敏感性有限。总之,候选的cir-miRNA已被确定为潜在的生物标志物,包括miR-423-5p、miR-93-5p和miR-4532。通过 miR-4532 进行筛选,并通过 miR-423-5p、miR-93-5p 进行确认,可用于鉴别 O. viverrini 流行区的 CCA 患者。
Differential circulating miRNA profiles identified miR-423-5p, miR-93-5p, and miR-4532 as potential biomarkers for cholangiocarcinoma diagnosis.
Background: Cholangiocarcinoma (CCA) is high in morbidity and mortality rates which may be due to asymptomatic and effective diagnostic methods not available. Therefore, an effective diagnosis is urgently needed.
Methods: Investigation of plasma circulating miRNA (cir-miRNA) was divided into two phases, including the discovery phase (pooled 10 samples each from three pools in each group) and the validation phase (17, 16, and 35 subjects of healthy control (HC), O. viverrini (OV), and CCA groups, respectively). The plasma from healthy control subjects, O. viverrini infected subjects, and CCA subjects was used. In the discovery phase, plasma was pooled by adding an equal volume of plasma, and cir-miRNA was isolated and analyzed with the nCounter® SPRINT Profiler. The significantly different cir-miRNAs were selected for the validation phase. In the validation phase, cir-miRNA was isolated and analyzed using real time-quantitative polymerase chain reaction (RT-qPCR). Subsequently, statistical analysis was conducted, and diagnostic parameters were calculated.
Results: Differential plasma cir-miRNA profile showed at least three candidates including miR-423-5p, miR-93-5p, and miR-4532 as potential biomarkers. From validation of these cir-miRNAs by RT-qPCR, the result showed that the satisfied sensitivity and specificity to differential CCA group from HC and OV group was obtained from miR-4532 (P < 0.05) while miR-423-5p and miR-93-5p can be used for differential CCA from OV and HC group (P < 0.05) with high specificity but limited the sensitivity. In conclusion, candidate cir-miRNAs have been identified as potential biomarkers including miR-423-5p, miR-93-5p and miR-4532. Screening by miR-4532 and confirmed with miR-423-5p, miR-93-5p were suggested for differential CCA patients in the endemic area of O. viverrini.
期刊介绍:
PeerJ is an open access peer-reviewed scientific journal covering research in the biological and medical sciences. At PeerJ, authors take out a lifetime publication plan (for as little as $99) which allows them to publish articles in the journal for free, forever. PeerJ has 5 Nobel Prize Winners on the Board; they have won several industry and media awards; and they are widely recognized as being one of the most interesting recent developments in academic publishing.