High Sensitivity and Specificity Platform to Validate MicroRNA Biomarkers in Cancer and Human Diseases.

IF 3.6 Q2 BIOCHEMISTRY & MOLECULAR BIOLOGY
Anastassia Kanavarioti, M Hassaan Rehman, Salma Qureshi, Aleena Rafiq, Madiha Sultan
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引用次数: 0

Abstract

We developed a technology for detecting and quantifying trace nucleic acids using a bracketing protocol designed to yield a copy number with approximately ± 20% accuracy across all concentrations. The microRNAs (miRNAs) let-7b, miR-15b, miR-21, miR-375 and miR-141 were measured in serum and urine samples from healthy subjects and patients with breast, prostate or pancreatic cancer. Detection and quantification were amplification-free and enabled using osmium-tagged probes and MinION, a nanopore array detection device. Combined serum from healthy men (Sigma-Aldrich, St. Louis, MO, USA #H6914) was used as a reference. Total RNA isolated from biospecimens using commercial kits was used as the miRNA source. The unprecedented ± 20% accuracy led to the conclusion that miRNA copy numbers must be normalized to the same RNA content, which in turn illustrates (i) independence from age, sex and ethnicity, as well as (ii) equivalence between serum and urine. miR-21, miR-375 and miR-141 copies in cancers were 1.8-fold overexpressed, exhibited zero overlap with healthy samples and had a p-value of 1.6 × 10-22, tentatively validating each miRNA as a multi-cancer biomarker. miR-15b was confirmed to be cancer-independent, whereas let-7b appeared to be a cancer biomarker for prostate and breast cancer, but not for pancreatic cancer.

验证癌症和人类疾病中 MicroRNA 生物标记物的高灵敏度和特异性平台。
我们开发了一种检测和量化痕量核酸的技术,该技术采用括弧协议,在所有浓度下产生的拷贝数精确度约为± 20%。我们测定了健康人和乳腺癌、前列腺癌或胰腺癌患者血清和尿液样本中的微量核糖核酸(miRNA)let-7b、miR-15b、miR-21、miR-375 和 miR-141。使用锇标记探针和纳米孔阵列检测设备 MinION 进行无扩增检测和定量。健康男性的混合血清(Sigma-Aldrich,St. Louis,MO,USA #H6914)用作参照物。使用商业试剂盒从生物样本中分离出的总 RNA 用作 miRNA 源。前所未有的 ± 20% 精确度使我们得出结论,miRNA 拷贝数必须归一化为相同的 RNA 含量,这反过来又说明了 (i) 与年龄、性别和种族无关,以及 (ii) 血清和尿液之间的等效性。miR-15b 被证实与癌症无关,而 let-7b 似乎是前列腺癌和乳腺癌的癌症生物标志物,但不是胰腺癌的生物标志物。
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来源期刊
Non-Coding RNA
Non-Coding RNA Biochemistry, Genetics and Molecular Biology-Genetics
CiteScore
6.70
自引率
4.70%
发文量
74
审稿时长
10 weeks
期刊介绍: Functional studies dealing with identification, structure-function relationships or biological activity of: small regulatory RNAs (miRNAs, siRNAs and piRNAs) associated with the RNA interference pathway small nuclear RNAs, small nucleolar and tRNAs derived small RNAs other types of small RNAs, such as those associated with splice junctions and transcription start sites long non-coding RNAs, including antisense RNAs, long ''intergenic'' RNAs, intronic RNAs and ''enhancer'' RNAs other classes of RNAs such as vault RNAs, scaRNAs, circular RNAs, 7SL RNAs, telomeric and centromeric RNAs regulatory functions of mRNAs and UTR-derived RNAs catalytic and allosteric (riboswitch) RNAs viral, transposon and repeat-derived RNAs bacterial regulatory RNAs, including CRISPR RNAS Analysis of RNA processing, RNA binding proteins, RNA signaling and RNA interaction pathways: DICER AGO, PIWI and PIWI-like proteins other classes of RNA binding and RNA transport proteins RNA interactions with chromatin-modifying complexes RNA interactions with DNA and other RNAs the role of RNA in the formation and function of specialized subnuclear organelles and other aspects of cell biology intercellular and intergenerational RNA signaling RNA processing structure-function relationships in RNA complexes RNA analyses, informatics, tools and technologies: transcriptomic analyses and technologies development of tools and technologies for RNA biology and therapeutics Translational studies involving long and short non-coding RNAs: identification of biomarkers development of new therapies involving microRNAs and other ncRNAs clinical studies involving microRNAs and other ncRNAs.
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