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一种基于碲化镉量子点不同热响应聚集/解聚的微小RNA检测方法。

An approach toward miRNA detection different thermo-responsive aggregation/disaggregation of CdTe quantum dots.

作者信息

Borghei Yasaman Sadat, Hosseini Morteza

机构信息

Department of Life Science Engineering, Faculty of New Sciences & Technologies, University of Tehran Tehran Iran

出版信息

RSC Adv. 2018 Aug 24;8(53):30148-30154. doi: 10.1039/c8ra04257h.

DOI:10.1039/c8ra04257h
PMID:35546848
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC9085396/
Abstract

MicroRNA-155 regulates the expression of 147 target genes that are involved in cancer pathways, and its expression level has been shown to be up-regulated in breast cancer. Thus, it is necessary to investigate the value of microRNA-155 for early diagnosis and prognosis of breast cancer. Here we present a novel and "light shift" spectral method for the detection of miRNA based on different thermo-responsive aggregation/disaggregation of CdTe quantum dots (CdTe QDs) by using single stranded DNA or a DNA/RNA heteroduplex as a template after heat treatment. In this method upon addition of the DNA/RNA heteroduplex, the CdTe QDs aggregate strongly due to their strong interaction with the double stranded nucleic acid, which results in fluorescence quenching. By applying the melting temperature ( ), the DNA/RNA heteroduplex denatures and two strands are dissociated, which disaggregates the QDs, effectively switching to fluorescence emission of QDs. These processes were investigated with Atomic Force Microscopy (AFM) and fluorescence spectroscopy. The proposed method has been used also for the determination of miR-155 in total RNAs extracted from the human breast carcinoma SK-BR-3 cells and normal human embryonic kidney cell line (HEK 293).

摘要

微小RNA-155调控147个参与癌症相关通路的靶基因的表达，且其表达水平在乳腺癌中呈上调。因此，有必要研究微小RNA-155在乳腺癌早期诊断和预后方面的价值。在此，我们提出一种基于碲化镉量子点（CdTe QDs）不同热响应聚集/解聚的新型“光移”光谱法，用于检测微小RNA。该方法在热处理后，以单链DNA或DNA/RNA异源双链体为模板。在此方法中，加入DNA/RNA异源双链体后，CdTe QDs由于与双链核酸的强相互作用而强烈聚集，导致荧光猝灭。通过应用解链温度（ ），DNA/RNA异源双链体变性且两条链解离，使量子点解聚，有效地转变为量子点的荧光发射。这些过程通过原子力显微镜（AFM）和荧光光谱进行了研究。所提出的方法还用于测定从人乳腺癌SK-BR-3细胞和正常人胚肾细胞系（HEK 293）中提取的总RNA中的miR-155。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/2512/9085396/a45f446f7afa/c8ra04257h-f5.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/2512/9085396/04b571735bcb/c8ra04257h-s1.jpg
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https://cdn.ncbi.nlm.nih.gov/pmc/blobs/2512/9085396/5f86bbb28133/c8ra04257h-f1.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/2512/9085396/48c816b8b67a/c8ra04257h-f2.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/2512/9085396/f079bc693555/c8ra04257h-f3.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/2512/9085396/8e4dccaf9fda/c8ra04257h-f4.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/2512/9085396/a45f446f7afa/c8ra04257h-f5.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/2512/9085396/04b571735bcb/c8ra04257h-s1.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/2512/9085396/69e96fc2975b/c8ra04257h-s2.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/2512/9085396/920c6a29f340/c8ra04257h-s3.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/2512/9085396/5f86bbb28133/c8ra04257h-f1.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/2512/9085396/48c816b8b67a/c8ra04257h-f2.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/2512/9085396/f079bc693555/c8ra04257h-f3.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/2512/9085396/8e4dccaf9fda/c8ra04257h-f4.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/2512/9085396/a45f446f7afa/c8ra04257h-f5.jpg

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