MICROCHIMICA ACTA
·
2014
·Vol. 182
·期 1
·241-248
·
IF 3.719
Wang Mo,Yang Zhiqing,Guo Yunlong,Wang Xinxu,Yin Huanshun,Ai Shiyun
We demonstrate a photo-electrochemical biosensor for the sensitive and specific detection of microRNA using Bi 2 S 3 nanorods as a photoactive material and streptavidin as the unit that inhibits photocurrent. Bi 2 S 3 nanorods were synthesized hydrothermally in organic phase and displayed excellent light-to-current conversion efficiency. The Bi 2 S 3 was deposited on an indium tin oxide (ITO) slice and then modified with gold nanoparticles onto which biotinylated hairpin probe DNA was deposited as a monolayer. Following hybridization between the biotinylated probe DNA and the target microRNA, the stem-loop structure of the probe DNA was unfolded and the biotin directed outwards into the solution. Streptavidin was then added to bind to biotin via the strong streptavidin-biotin interactions. This causes the photocurrent of the modified ITO to decrease due to steric hindrance that blocks the transfer of electrons from added ascorbic acid to the surface of the electrode. The method has a detection limit as low as 3.5 fM of microRNA and can excellently discriminate even singly mismatched microRNA. The method was successfully applied to investigate the effect of abscisic acid on the expression level of microRNA-159a in seeds of Arabidopsis thaliana . We conclude that the assay presented here has a large potential as a method for quantification of microRNA and for studying the epigenetic regulation of flowering plants.
The photocurrent generation and decrease mechanism of the PEC biosensor.
研究领域:
生物传感器
·
光化学
·
纳米技术
·
分子生物学
·
植物科学
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Oncology Letters
·
2021
·Vol. 21
·期 3
·1-1
·
IF 2.967
Huizhe Zheng,Bin Yan,Qi Wu,Jingli Zhang
Chemotherapy drug 5‑fluorouracil (5‑FU) is the first‑line treatment for colorectal cancer (CRC); however, 5‑FU resistance decreases CRC therapeutic efficiency. A previous study revealed that microRNA (miR)‑9‑5p serves an antitumor effect in CRC. However, the effect of miR‑9‑5p in CRC chemoresistance remains unknown. In the present study, two CRC cell lines, including HT‑29 and HCT‑116 cells, were used to investigate the impact of miR‑9‑5p in overcoming 5‑FU resistance. The results revealed that treatment with 5‑FU decreased CRC cell viability and upregulated miR‑9‑5p expression in both CRC cells. Knockdown of miR‑9‑5p decreased HCT‑116 cell sensitivity to 5‑FU and inhibited apoptosis. By contrast, miR‑9‑5p overexpression enhanced the sensitivity of HT‑29 cells to 5‑FU and induced apoptosis. Additionally, it was confirmed that miR‑9‑5p directly targeted high mobility group A2 (HMGA2). HMGA2 overexpression reversed miR‑9‑5p‑induced HT‑29 apoptosis. The present study indicated that miR‑9‑5p enhanced the sensitivity of CRC cells to 5‑FU via downregulating HMGA2 expression.
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ANALYTICAL CHEMISTRY
·
2021
·
IF 6.986
Jiamin Wu,Xia Zhou,Ping Li,Xiaoling Lin,Jinhua Wang,Ziwei Hu,Pengcheng Zhang,Dong Chen,Huaihong Cai,Reinhard Niessner,Christoph Haisch,Pinghua Sun,Yun Zheng,Zhengjin Jiang,Haibo Zhou
Sensitive and simultaneous detection of multiple cancer-related biomarkers in serum is essential for diagnosis, therapy, prognosis, and staging of cancer. Herein, we proposed a magnetically assisted sandwich-type surface-enhanced Raman scattering (SERS)-based biosensor for ultrasensitive and multiplex detection of three hepatocellular carcinoma-related microRNA (miRNA) biomarkers. The biosensor consists of an SERS tag (probe DNA-conjugated DNA-engineered fractal gold nanoparticles, F-AuNPs) and a magnetic capture substrate (capture DNA-conjugated Ag-coated magnetic nanoparticles, AgMNPs). The proposed strategy achieved simultaneous and sensitive detection of three miRNAs (miRNA-122, miRNA-223, and miRNA-21), and the limits of detection of the three miRNAs in human serum are 349 aM for miRNA-122, 374 aM for miRNA-223, and 311 aM for miRNA-21. High selectivity and accuracy of the SERS biosensor were proved by practical analysis in human serum. Moreover, the biosensor exhibited good practicability in multiplex detection of three miRNAs in 92 clinical sera from AFP-negative patients, patients before and after hepatectomy, recurred and relapse-free patients after hepatectomy, and hepatocellular carcinoma patients at distinct Barcelona clinic liver cancer stages. The experiment results demonstrate that our SERS-based assay is a promising candidate in clinical application and exhibited potential for the prediction, diagnosis, monitoring, and staging of cancers.
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MICROCHIMICA ACTA
·
2021
·Vol. 188
·期 1
·1-12
·
IF 5.833
Yang Lili,Wang Jiasheng,Lü Haitao,Hui Ni
A dual-sensing platform is proposed based on multi-walled carbon nanotubes/Prussian blue–functionalized polypyrrole nanowire array (PPY/MWCNTs/PB). Highly aligned PPY nanowire arrays were electrochemically prepared on the surface of glassy carbon electrodes, which were doped with MWCNTs/PB nanocomposites. The nanomaterial combines the characteristics of the PPY nanowires (high conductivity and large specific surface area) and MWCNTs/PB (excellent catalytic performance and intrinsic redox activity). Owing to the nanowire microstructure and outstanding electrical properties, the PPY/MWCNTs/PB nanowire arrays show excellent electrocatalysis of the reduction of hydrogen peroxide and facilitate the construction of a high-performance biosensing platform for microRNA (miRNA). A linear relationship between analytical signal and concentration of hydrogen peroxide and miRNA was obtained in the range 5 to 503 µM (1.4–5.1 mM) and 0.1 pM to 1 nM, and detection limits of 1.7 μM and 33.4 fM, respectively. This new supersensitive sensing platform has broad application prospects of biomolecule and other analyte determination in drug, biomedical, plant protection, and environmental analysis. Graphical abstract
Prussian blue/multi-walled carbon nanotubes functionalized polypyrrole nanowire arrays (PPY/MWCNTs/PB) were prepared by a facile one-step electrochemical method. PPY/MWCNTs/PB nanowire arrays show excellent electrocatalysis of the reduction of H 2 O 2 and facilitate the construction of a high-performance biosensing platform for microRNA.
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ENVIRONMENTAL SCIENCE AND POLLUTION RESEARCH
·
2020
·Vol. 27
·期 14
·17290-17302
·
IF 3.056
Ma Lin,Yu Haiyang,Wang Xu,Li Dan,Zhang Yumin,Pei Xiucong,Duan Zhiwen,Ma Mingyue
The effect of prenatal bisphenol A (BPA) exposure is increasingly concerned. We investigated the effect of maternal BPA exposure during pregnancy on male offspring and its potential mechanism. Thirty pregnant Sprague Dawley (SD) rats were randomly divided into exposed and control groups. At PND56, the number of sperm, luteinizing hormone, and testosterone in the BPA-exposed group decreased, and testicular tissue structure was damaged in offsprings. At GD20, the miRNA profile in the testis of male offspring was examined and the expression levels of 28 deregulated miRNAs were validated by qRT-PCR. We found that miR-361-5p , miR-203a-3p , and miR-19b-2-5p had significantly different expression levels in the testis. These results suggest that maternal exposure to BPA can lead to differential changes in progeny miRNAs, which will provide direction for future in-depth mechanisms of reproductive injury.
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TALANTA
·
2021
·
IF 6.057
Yuanyuan Yao,Hongding Zhang,Tongtong Tian,Yixin Liu,Rendan Zhu,Ji Ji,Baohong Liu
With the emergence of microRNA (miRNA) as a key player in early clinical disease diagnosis, development of rapidly sensitive and quantitative miRNA detection methods are imperative. Herein, a label-free SERS assay coupled with duplex-specific nuclease (DSN) signal amplification strategy was proposed for facilely ultrasensitive and quantitative analysis of miRNA-21. Firstly, magnetic beads assembled with excessive capture DNA were utilized to hybridize the target miRNA-21. These DNA-RNA heteroduplexes were cleaved by DSN to generate small nucleotide fragments into the supernatant and the miRNA-21 released and rehybridized another DNA, going to the next DSN cycle. Consequently, numerous of small nucleotide fragments of capture DNA were released from magnetic beads and the miRNA-21 signal was transferred and amplified by the SERS signals of total phosphate backbones which are abundant in nucleotide. Furthermore, iodide-modified Ag nanoparticles (AgINPs) was employed to generate a strong and reproducible SERS signal. The proposed method displayed excellent performance for miRNA-21 detection with the linear range from 0.33 fM to 3.3 pM, and a lower detection limit of 42 aM. Moreover, this strategy exhibited effectively base discrimination capability and was successfully applied for monitoring the expression levels of miRNA-21 in different cancer cell lines and human serum.
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SENSORS AND ACTUATORS B-CHEMICAL
·
2018
·
IF 5.667
Weipan Peng,Qian Zhao,Jiafang Piao,Miao Zhao,Yiwen Huang,Bo Zhang,Weichen Gao,Dianming Zhou,Guiming Shu,Xiaoqun Gong,Jin Chang
The detection and analysis of microRNA (miRNA) can provide critical information for biomedical research and clinical diagnosis. In this paper, we constructed a simple method to achieve ultra-sensitive and rapid detection of miRNA based on the double-specific nuclease (DSN) assisting target recovery and horseradish peroxidase (HRP) cascading signal amplification. In the existence of target microRNA-21 (miR-21), the DSN enzyme specifically cut the single-stranded DNA (ssDNA) hybridized with the target miRNA but still kept the target miRNA intact, so there were more free HRP released from the polystyrene microparticles-ssDNA-HRP conjugates (PS-ssDNA-HRP) because of the target recycle and signal amplification. The free HRP in the supernatant were collected to react with the substrate 3,3',5,5'-tetramethylbenzidine (TMB). The color and absorbance changes of HRP-TMB system further resulted in qualitative screening by naked eyes and quantitative detection by the microplate reader with the detection limit as low as 3.1 fM. This assay protocol not only showed high sensitivity and selectivity, but also well distinguished the difference between the miRNA family members, as well as the simple operation, fast detection, naked eye qualitative and microplate reader quantification. It has great application value in the biomedical research and clinical diagnosis.
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ANALYTICA CHIMICA ACTA
·
2018
·
IF 5.123
Fengzhou Xu,Lan Luo,Hui Shi,Xiaoxiao He,Yanli Lei,Jinlu Tang,Dinggeng He,Zhenzhen Qiao,Kemin Wang
Poly(thymine)-hosted copper nanoparticles (poly T-CuNPs) have emerged as a promising label-free fluorophore for bioanalysis, but its application in RNA-related studies is still rarely explored. Herein, by utilizing duplex-specific nuclease (DSN) as a convertor to integrate target recycling mechanism into terminal deoxynucleotidyl transferase (TdT)-mediated superlong poly T-CuNPs platform, a specific and sensitive method for microRNA detection has been developed. In this strategy, a 3′-phosphorylated DNA probe can hybridize with target RNA and then be cut by DSN to produce 3′-hydroxylated fragments, which can be further tailed by TdT with superlong poly T for fluorescent CuNPs synthesis. As proof of concept, an analysis of let-7d was achieved with a good linear correlation between 20 and 1000 pM (R 2 = 0.9965) and a detection limit of 20 pM. Moreover, both homologous and heterologous microRNAs were also effectively discriminated. This strategy might pave a brand-new way for designing label-free and sensitive microRNA assays.
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SENSORS AND ACTUATORS B-CHEMICAL
·
2017
·
IF 5.401
Bing Wang,Yu-Xiang Dong,Yu-Ling Wang,Jun-Tao Cao,Shu-Hui Ma,Yan-Ming Liu
A novel and simple photoelectrochemical (PEC) biosensing method for microRNA-21 (miRNA-21) detection is reported based on energy transfer (ET) between CdS:Mn doped structure (CdS:Mn) and Au nanoparticles (AuNPs). In this protocol, TiO 2 -CdS:Mn hybrid structure was used as a sensing platform for hairpin DNA immobilization. In the absence of miRNA-21, the immobilized DNA was in the hairpin form. In this state, the photocurrent of the electrode was greatly depressed, due to the effective ET effect produced by short interparticle distance between CdS:Mn and AuNPs. In the presence of miRNA-21, the hairpin DNA hybridized with miRNA-21 and changed into a more rigid, rodlike double helix, which forced the AuNPs away from the electrode surface, leading to obvious recover of photocurrent because of the vanished damping effect. Integrating the fine PEC performance of TiO 2 -CdS:Mn hybrid structure with the significant ET effect between CdS:Mn and AuNPs, the sensitive detection of miRNA-21 was realized in a linear range of 1.0 fM to 10.0 pM with a low detection limit of 0.5 fM. This method might be aussichtsreich for the detection of miRNAs and other biomarkers.
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JOURNAL OF CELLULAR BIOCHEMISTRY
·
2018
·Vol. 120
·期 6
·9147-9158
·
IF 2.959
Ya-Jun Song,Jun-Ying Cao,Zhuang Jin,Wen-Gang Hu,Rong-Hua Wu,Lu-Hai Tian,Bo Yang,Jin Wang,Ya Xiao,Chi-Bing Huang
Interstitial cystitis (IC) is a heterogeneous syndrome with unknown etiology, and microRNAs (miRs) were found to be involved in IC. In our study, we aim to explore the role of miR-132 in the inflammatory response and detrusor fibrosis in IC through the Janus kinase-signal transducer and activator of transcription (JAK-STAT) signaling pathway in rat models. A rat model of IC was established and treated with the miR-132 mimic, miR-132 inhibitor, and/or JAK-STAT signaling pathway inhibitor AG490. Enzyme-linked immunosorbent assay was applied to measure the expression of interleukin (IL)-6, IL-10, interferon-γ (IFN-γ), and tumor necrosis factor-α (TNF-α), and intercellular adhesion molecule-1 (ICAM-1). The urodynamic test was performed to assess urodynamic parameters, and reverse transcription quantitative polymerase chain reaction and Western blot analysis for the expression of miR-132, STAT4, suppressors of cytokine signaling 3 (SOCS3), JAK2, vascular endothelial growth factor (VEGF), IFN-γ, and TNF-α. IC rats treated with miR-132 inhibitor and AG490 had decreased collagen fiber, inflammatory cell infiltration, and mast cells, lower expression of IL-6, IL-10, IFN-γ, TNF-α, ICAM-1, collagens I and III, and alleviated urodynamic parameters and decreased expression of STAT4, VEGF, JAK2, IFN-γ, TNF-α, and increased expression of SOCS3. Taken together, our data indicate that downregulation of miR-132 alleviates inflammatory response and detrusor fibrosis in IC via the inhibition of the JAK-STAT signaling pathway.
研究领域:
泌尿学
·
分子生物学
·
药理学
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ACS Sensors
·
2024
·
IF 8.2
Rongrong Huang,Peipei Wan,Shengjie Hu,Chenyang Zhang,Wenjun Miao
MicroRNAs (miRNAs) in dermal interstitial fluid (ISF) have recently been recognized as clinically promising biomarkers for the diagnosis and prognosis of cancer. However, the detection poses significant challenges, primarily due to the low abundance of miRNAs and the limitations of current sampling techniques. To address this issue, we develop novel porous microneedles (PMNs) array-based sensor composed of poly(vinyl alcohol) porous hydrogel and DNA-templated silver nanoclusters (AgNCs) to facilitate the enrichment and highly sensitive detection of ISF miRNA. Leveraging the capillary action facilitated by its unique porous structure and the swelling properties of the hydrogel, the PMNs array can efficiently extract 2.7 ± 0.3 mg of ISF within 5 min. Additionally, the interconnected pores within the PMNs array contribute to an increased specific surface area, thereby offering a convenient platform for the decoration of DNA-templated AgNCs. The immobilized large amount of AgNCs effectively capture the target miRNA from the extracted ISF, resulting in miRNA-induced fluorescence quenching of AgNCs. Subsequently, the introduction of the duplex-specific nuclease leads to the cleavage of DNA in DNA-RNA heteroduplexes, which release miRNA to interact with other AgNCs. This process of target recycling triggers a further reduction in fluorescence intensity, thereby enabling sensitive detection of the low-abundant miRNA down to 1.6 pM. Both in vitro and in vivo experiments validate the efficacy of the AgNCs immobilized PMNs array for the detection of miRNA biomarkers in ISF within minutes. These results indicate that the proposed PMNs array-based sensor holds great potential for the development of noninvasive personalized diagnostic strategies.
研究领域:
生物医学工程
·
分子诊断
·
纳米技术
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Journal of Analytical Science and Technology
·
2026
·Vol. 17
·期 1
·12
·
IF 2.4
Chen Lingling,Chen Chen
Non-small cell lung cancer (NSCLC) is a leading cause of global cancer mortality, with a poor five-year survival rate primarily attributed to late diagnosis and therapeutic resistance. Emerging evidence highlights the critical role of microRNAs (miRNAs) in NSCLC pathogenesis, progression, and prognosis. However, conventional polymerase chain reaction (PCR)-based miRNA detection methods often lack sufficient sensitivity and robustness. This study presents a novel, self-assembled bio-sensing platform for the highly sensitive analysis of circulating NSCLC-associated miRNAs. The platform integrates a target-primed hybridization chain reaction (THSP) with cooperative self-primed circular amplification, utilizing a Variable Hairpin Probe (VHP) biosensor coupled with the SYBR Green I fluorescent dye system. Upon specific recognition of target miRNA-21, the system initiates a cascade amplification, minimizing nonspecific background. The activated reaction generates abundant double-stranded DNA products, which bind SYBR Green I to produce a strong fluorescent signal without requiring expensive labeled probes. This straightforward method achieves a detection limit of 1.33 fM, significantly enhancing the sensitivity and practicality. It offers a promising strategy for developing highly specific nucleic acid analysis tools, advancing non-invasive miRNA detection toward precise and personalized NSCLC management.
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