新型二硫化钼荧光传感器检测乳腺癌相关miRNA

doi:10.3969/j.issn.1673-8640.2017.05.018

检验医学 ›› 2017, Vol. 32 ›› Issue (5): 427-430.DOI: 10.3969/j.issn.1673-8640.2017.05.018

• 技术研究与评价·论著 • 上一篇下一篇

新型二硫化钼荧光传感器检测乳腺癌相关miRNA

蔡冰洁¹, 张国军², 李艳¹

1.武汉大学人民医院检验科,湖北武汉 430060
2.湖北中医药大学,湖北武汉 430065

收稿日期:2016-03-01 出版日期:2017-05-20 发布日期:2017-06-06
作者简介:null
作者简介：蔡冰洁,女,1989年生,硕士,主要从事纳米生物传感技术研究。
基金资助:
国家自然科学基金项目（21475034）

MoS₂-based fluorescence sensor for the detection of breast cancer-related microRNA

CAI Bingjie¹, ZHANG Guojun², LI Yan¹

1. Department of Clinical Laboratory,Renmin Hospital of Wuhan University,Wuhan 430060,Hubei,China
2. Hubei University of Chinese Medicine,Wuhan 430065,Hubei,China

Received:2016-03-01 Online:2017-05-20 Published:2017-06-06

摘要/Abstract

摘要：

目的建立一种快速简便、高敏感性、高特异性的基于新型纳米材料二硫化钼的荧光传感器用于乳腺癌相关微小RNA（miRNA）检测的方法。方法在荧光标记的DNA探针体系中加入新型纳米材料二硫化钼构建一种荧光传感器,然后在荧光传感器中分别加入非互补miRNA、单碱基错配的miRNA、不同浓度的互补miRNA与荧光标记的DNA探针进行杂交,通过检测杂交前后荧光信号的变化来检测miRNA。结果检测只需40 min即可完成,对互补miRNA的检出限可达pmol/L级,且能够将互补、非互补、单碱基错配的miRNA区分开来。结论该新型的纳米材料二硫化钼荧光传感器分析速度快、敏感性和特异性高,建立的检测方法可应用于乳腺癌相关miRNA的快速检测,是一种非常有潜力的检测方法。

关键词: 二硫化钼, 荧光传感器, 乳腺癌, 微小RNA

Abstract:

Objective To establish a fast,convenient,sensitive and specific fluorescence sensor based on novel nanomaterial MoS₂ to detect breast cancer-related microRNA（miRNA）. Methodse The fluorescence sensor was fabricated by mixing novel nanomaterial MoS₂ with fluorescent dye-labeled DNA probe. Noncomplementary miRNA,one-base mismatched miRNA and different concentrations of complementary miRNA were introduced to fluorescence sensor for hybridization with DNA probe. The miRNA was detected through monitoring fluorescence signal change before and after DNA probe-miRNA hybridization. Results The pmol/L level of complementary miRNA could be detected within 40 min,and fluorescence sensor could discriminate complementary miRNA from noncomplementary miRNA and one-base mismatched miRNA. Conclusions The novel nanomaterial MoS₂-based fluorescence sensor has the advantages of fast,high sensitivity and specificity,and it is potential for the detection of breast cancer-related miRNA.

Key words: MoS2, Fluorescence sensor, Breast cancer, MicroRNA

中图分类号:

R446.1

蔡冰洁, 张国军, 李艳. 新型二硫化钼荧光传感器检测乳腺癌相关miRNA[J]. 检验医学, 2017, 32(5): 427-430.

CAI Bingjie, ZHANG Guojun, LI Yan. MoS₂-based fluorescence sensor for the detection of breast cancer-related microRNA[J]. Laboratory Medicine, 2017, 32(5): 427-430.

图/表 5

表1 荧光标记的探针DNA及miRNA序列

核酸	序列（5'~3'）	碱基数
荧光标记DNA探针	TCAACATCAGTCTGATAAGCTA-FAM	22
miRNA-21序列	UAGCUUAUCAGACUGAUGUUGA	22
单碱基错配miRNA	UAGCUUAUCAGACUGAAGUUGA	22
非互补miRNA	AGAGGAUGUUAGUAAGGUGGUU	22

图1 新型纳米材料二硫化钼荧光传感器检测miRNA的原理示意图

图2 方法可行性验证荧光光谱图

图3 荧光信号随反应时间变化监测图

图4 不同浓度miRNA-21 加入体系反应后得到的标准荧光曲线（a）及线性曲线（b）

参考文献 14

[1]	石建伟,唐智柳,蔡美玉,等. 2008~2012年我国女性乳腺癌流行状况的系统性综述[J]. 中国妇幼保健,2014,29(10):1622-1626.
[2]	魏旭倩,王伟祥,王学锋,等. 外周血CEA mRNA、Survivin mRNA、MMP-2 mRNA和TIMP-2 mRNA水平与乳腺癌的关系[J]. 检验医学,2011,26(2):94-99.
[3]	GAO J,ZHANG Q,XU J,et al.Clinical significance of serum miR-21 in breast cancer compared with CA153 and CEA[J]. Chin J Cancer Res,2013,25(6):743-748.
[4]	王卉,刘水逸,吴唐维,等. miR-21 在乳腺癌中的表达及其对乳腺癌侵袭转移的影响[J]. 中华检验医学杂志,2015,38(3):186-190.
[5]	PETROVIĆ N,MANDUŠIĆ V,STANOJEVIĆ B,et al. The difference in miR-21 expression levels between invasive and non-invasive breast cancers emphasizes its role in breast cancer invasion[J]. Med Oncol,2014,31(3):867.
[6]	KUMAR S,KEERTHANA R,PAZHANIMUTHU A,et al.Overexpression of circulating miRNA-21 and miRNA-146a in plasma samples of breast cancer patients[J]. Indian J Biochem Biophys,2013,50(3):210-214.
[7]	LIU J,LU Y.A DNAzyme catalytic beacon sensor for paramagnetic Cu2+ ions in aqueous solution with high sensitivity and selectivity[J]. J Am Chem Soc,2007,129(32):9838-9839.
[8]	ZHU C,ZENG Z,LI H,et al.Single-layer MoS2-based nanoprobes for homogeneous detection of biomolecules[J]. J Am Chem Soc,2013,135(16):5998-6001.
[9]	RAFIEE-POUR H A,BEHPOUR M,KESHAVARZ M. A novel label-free electrochemical miRNA biosensor using methylene blue as redox indicator:application to breast cancer biomarker miRNA-21[J]. Biosens Bioelectron,2016,77:202-207.
[10]	CROCE C M.Causes and consequences of microRNA dysregulation in cancer[J]. Nat Rev Genet,2009,10(10):704-714.
[11]	BOERI M,VERRI C,CONTE D,et al.MicroRNA signatures in tissues and plasma predict development and prognosis of computed tomography detected lung cancer[J]. Proc Natl Acad Sci U S A,2011,108(9):3713-3718.
[12]	IORIO M V,CROCE C M.MicroRNA dysregulation in cancer:diagnostics,monitoring and therapeutics. A comprehensive review[J]. EMBO Mol Med,2012,4(3):143-159.
[13]	GUO S,YANG F,ZHANG Y,et al.Amplified fluorescence sensing of miRNA by combination of graphene oxide with duplex-specific nuclease[J]. Anal Methods,2014,6(11):3598.
[14]	XI Q,ZHOU D M,KAN Y Y,et al.Highly sensitive and selective strategy for microRNA detection based on WS2 nanosheet mediated fluorescence quenching and duplex-specific nuclease signal amplification[J]. Anal Chem,2014,86(3):1361-1365.

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新型二硫化钼荧光传感器检测乳腺癌相关miRNA

MoS₂-based fluorescence sensor for the detection of breast cancer-related microRNA

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新型二硫化钼荧光传感器检测乳腺癌相关miRNA

MoS2-based fluorescence sensor for the detection of breast cancer-related microRNA

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MoS₂-based fluorescence sensor for the detection of breast cancer-related microRNA