BIOSENSORS & BIOELECTRONICS
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2019
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IF 9.518
Lili Zhou,Ting Wang,Yan Bai,Yi Li,Juhui Qiu,Wen Yu,Sheng Zhang
s Rapid and efficient detection of tumor marker at the early stages is one of the crucial challenges in cancer diagnostics and therapy. In this study, an ultrasensitive electrochemical biosensor was fabricated by dual-amplified strategy for the detection of ultra-trace microRNA-141 (miRNA-141). Firstly, two split sequences contained G-quadruplex were connected by click chemistry-mediated nucleic acid strands self-assembly and the obtained complete G-quadruplex was complementary with miRNA-141 to formed DNA-RNA hybrid duplexes. Subsequently, the formed DNA-RNA hybrid duplexes were specifically recognized by duplex-specific nuclease (DSN), and the DNA part of the duplexes were cleaved and the miRNA-141 were released to trigger next cycle, which acquired a primal signal amplification by enzyme-assisted target recycling (EATR). Moreover, amino and thiol group multi-labeled functionalized fullerene nanoparticles (FC60) with a larger surface active sites and better biocompatibility, were designed rationally to modify the Au electrodes, which produced multiply-enhanced amplified signal. This dual-amplified sensing system exhibited a remarkable analytical performance for the detection of miRNA-141 in concentrations ranging from 0.1 pM to 100 nM and the detection limit of 7.78 fM was obtained. Compared with the biosensor with single amplification strategy such as EATR, this electrochemical biosensor based on dual-amplified strategy exhibited an excellent discrimination capability and higher analytical performance. Therefore, this electrochemical biosensor might hold a great potential for further applications in biomedical research and early clinical diagnosis.
研究领域:
生物医学工程
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分子诊断学
·
癌症早期检测
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高影响因子
MICROCHIMICA ACTA
·
2018
·Vol. 185
·期 2
·1-8
·
IF 5.705
Jian Yannan,Wang He,Lan Feifei,Liang Linlin,Ren Na,Liu Haiyun,Ge Shenguang,Yu Jinghua
An electrochemiluminescence (ECL) biosensor is described for the detection of microRNA (miRNA-155) based on tris(bipyridine)ruthenium(II) functionalized metal organic framework (RuMOF) materials. The material was prepared by a solvothermal method and was found to be stable even in acidic solution. However, it is selectively and sensitively disassembled by Hg(II) ions, resulting in the release of large quantities of Ru(II)(bpy) 3 ions, which produces a strong ECL signal. In view of the ion-selective disassembly and release and strand displacement process, an ultrasensitive ECL sensing method was established for detection of microRNAs. In the presence of the target, the hairpin structure of H1 can open and hybridize with the hairpin probe H2 to form a more stable H1-H2 duplex structure than the H1-target hybrid. The target of hybridization to H1 was immediately freed from the structure and the released target re-entered the new hairpin assembly target recovery process. The remaining H2 single fragment can bind to the I-RuMOFs-conjugates. The more hairpin probes H1, the more I-RuMOFs-conjugates load the DNA fragments, leading to the signal amplification. The method works in the 0.8 f. to 1.0 nM miRNA-155 concentration range and has a detection limit of 0.3 fM. The assay is sensitive, fairly specific and remarkably stable. In our perception, it offers an attractive tool for the sensitive detection of microRNAs in clinical samples.
An electrochemiluminescence (ECL) based biosensor is described for the detection of microRNA (miRNA-155) based on the use of a metal organic framework functionalized with ruthenium(II)tris(bipyridine) that was deposited on a glassy carbon electrode (GCE) modified with gold nanoparticles.
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BIOSENSORS & BIOELECTRONICS
·
2023
·
IF 12.6
Chunyan Wang,Yaxing Zhang,Chang Liu,Siyu Gou,Shanjin Hu,Weiwei Guo
Point-of-care testing (POCT) platforms for microRNA (miRNA) detection have attracted considerable attention in recent years, due to the increasingly important role of miRNA as biomarkers for the diagnosis of many diseases, such as cancers. However, several limitations such as the requirement of enzyme-related amplification system, expensive preservation cost, sophisticated analysis instruments and tedious operations of conventional miRNA biosensing devices severely hinder their widespread applications. In this work, a portable and smart colorimetric analysis platform was developed by employing the ultrathin DNA-gold nanoparticle (AuNP) hybrid hydrogel film as the signaling unit and the enzyme-free entropy-driven dynamic DNA network (EDN) as the signal converter and amplification unit. By programming the DNA sequences of the EDN, the EDN could respond to a specific miRNA, with miRNA-155 or miRNA-21 as the model target, and release a converter DNA with amplified concentration to further trigger the release of AuNPs from the hydrogel film as a colorimetric signal output. To avoid the use of sophisticated spectral instruments, digital analysis based on primary three-color channel (R/G/B) was further introduced by using user-friendly camera and image processing software, and a detection limit at pM level was achieved. Moreover, by introducing H 2 O 2 -mediated AuNPs enlargement procedure in the colorimetric analysis platform, the detection limit for miRNA target could further be enhanced to fM level. The POCT platform is also portable and storable with a good storage stability for at least 45 days, suggesting its great potential in practical diagnosis applications.
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Bioengineered
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2021
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IF 3.269
Haixia Liu,Mei Song,Xiaoyan Sun,Xin Zhang,Huayan Miao,Yankui Wang
Cervical cancer (CC) is the fourth most common cancers among women worldwide. T-box transcription factor 1 (TBX1), a member of the T-box family, has anti-tumor effects in some types of cancer, but its role in CC is yet unknown. The aim of this study is to investigate the functions and underlying mechanisms of TBX1 in CC. Online database UALCAN showed that TBX1 was down-regulated in CC tissues compared with normal tissues and patients with lower TBX1 expression level had a poor prognosis. TBX1 overexpression significantly decreased the proliferation, migration, and invasion of Hela and SiHa cells. Conversely, cell apoptosis and chemosensitivity to cisplatin were promoted in TBX1-overexpressing CC cells. Moreover, up-regulation of TBX1 inhibited both AKT and MAPK signaling pathways. Furthermore, dual luciferase report assay indicated that TBX1 could directly bind to miR-6727-5p. In addition, TBX1 expression was inhibited by miR-6727-5p mimic and up-regulated by miR-6727-5p inhibitor. Knockdown of TBX1 reversed the inhibitory effect of the miR-6727-5p inhibitor on CC cells. This study demonstrates that TBX1, a target gene of miR-6727-5p, acts as a tumor suppressor in CC, indicating that TBX1 may be a new target for CC therapy.
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Journal of Oncology
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2022
·
IF 4.501
Weiwei Wang,Lipei Wu,Jiale Tian,Wenhui Yan,Chunrun Qi,Wuchao Liu,Shihai Xuan,Anquan Shang
Application of extracellular vesicles (EVs) for cancer treatment has been well-documented. We probed into the potential role of cervical cancer cells-secreted EVs by transferring miR-146a-5p in cervical cancer. After characterization of miR-146a-5p expression in clinical cervical cancer tissue samples, gain- and loss-of-function experiments were implemented to test the effect of miR-146a-5p on the invasion, epithelial-mesenchymal transition (EMT), and anoikis in cervical cancer cells. EVs were isolated from high-metastatic cervical cancer cells, after which their effects on the malignant behaviors of low-metastatic cervical cancer cells were assessed in a co-culture system. Luciferase assay was implemented to validate the putative binding relationship between miR-146a-5p and WWC2, followed by further investigation of downstream pathway (Hippo-YAP). Finally, nude mouse lung metastasis model was developed for in vivo validation. miR-146a-5p was elevated in cervical cancer tissues and high miR-146a-5p expression promoted the metastatic potential of cervical cancer cells through enhancing their invasiveness and anoikis resistance, and inducing EMT. Furthermore, miR-146a-5p carried by EVs secreted by highly metastatic cervical cancer cells could promote the metastasis of low-metastatic cervical cancer cells. Mechanistically, miR-146a-5p targeted WWC2 to activate YAP, by which it inhibited the phosphorylation of cofilin, and promoted the process of cofilin-mediated depolymerization of F-actin to G-actin. In vivo data demonstrated that EVs-carried miR-146a-5p promoted tumor metastasis through the WWC2/YAP axis. Cancer-derived EVs delivered pro-metastatic miR-146a-5p to regulate the actin dynamics in cervical cancer, thereby leading to cancer metastasis. This experiment highlighted an appealing therapeutic modality for cervical cancer.
研究领域:
肿瘤学
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分子生物学
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细胞生物学
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高影响因子
Analytical Methods
·
2019
·Vol. 11
·期 32
·4137-4145
·
IF 2.378
Feng Cui,Qianying Qiu,Gang Peng,Xiaoyan Li,Xi Liu,Xiaojun Chen
The microRNA family shows great promise for the diagnosis and prediction of different cancers as biomarkers, in which miRNA-155 (miR155) is recognized as a biomarker for lung cancer and thus has drawn much attention. Therefore, the development of specific and accurate methods to detect miR155 has become a potential research hotspot. In this work, we presented a fluorescence resonance energy transfer (FRET) technique by utilizing core–shell CdSeTe/ZnS quantum dots (QDs) as a signal reporter and a target recycling strategy as a signal amplifier for the detection of miR155. In the presence of miR155, part of the hairpin probe (HP) immobilized onto the surface of gold nanoparticles (AuNPs)@SiO2, could hybridize with it. Then, numerous H1-AuNPs@SiO2 could be produced with the aid of exonuclease III. With the linkage of the assistant chain A2, H1-AuNPs@SiO2 combined with A1-QDs and then the fluorescence (FL) signal of QDs was quenched by AuNPs@SiO2via FRET. The more miR155 present in solution, the weaker the detected FL signal. A linear response range of 0.01 fM to 1 nM and low detection limit of 3 aM were obtained using the proposed method. Furthermore, this method showed high specificity and satisfactory recovery between 93 and 105% in real human serum samples, illustrating its great potential for practical applications.
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TALANTA
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2021
·
IF 6.057
Cai-Xia Shi,Yan-Zi Li,Qing Liu,Zeng-Ping Chen,Sha-Sha Li,Ru-Qin Yu
Biosensors based on various spectroscopic techniques discriminate the target microRNA (miRNA) from non-target ones with single nucleotide polymorphisms (SNPs) according to the differences in signal intensities which can be caused by other factors besides SNPs. As a result, they are liable to produce false positive results. Herein, we report an attempt to develop a false-positive resistance, sensitive and reliable mass spectrometric platform for miRNA detection. In the proposed platform, the qualitative and quantitative information of the target miRNA was obtained through analyzing mass spectral responses of the multiply charged ions of the residual fragments of the probe DNA produced during exonuclease III assisted signal amplification reaction using an advanced data analysis method. The proposed platform could achieve sensitive and accurate quantitative results for the target miRNA (e.g., miRNA-141) in complex medium with a detection limit of about 1 pM, and unambiguously identify non-target miRNAs with SNPs based on the length distribution patterns of residual fragments of probe DNA. The findings obtained in this study might open an avenue for the design of new miRNA detection methods based on mass spectrometry in combination with various nuclease assisted signal amplification strategies.
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ELECTROCHIMICA ACTA
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2020
·
IF 6.215
Xiangyu Ma,Kun Qian,Onome Ejeromedoghene,Martha Kandawa-Schulz,Yihong Wang
MicroRNAs (miRNAs) are becoming biomarkers for the clinical diagnosis of many diseases. However, their intrinsic properties such as low enrichment and short sequence restrict their applications. Here, we designed an ultrasensitive dual signal amplification electrochemical biosensor based on sodium alginate-polypyrrole/Au nanoparticles (SA-PPy/AuNPs) nanocomposite for the detection of miRNAs. Firstly, a planar intercalated copper(II) complex was synthesized to be the signal molecule. Then, catalytic hairpin assembly (CHA) reaction and catalytic reaction between ferric ion and copper ion were combined as a dual signal amplification to improve the sensitivity of the biosensor. In the presence of target miRNAs, multiple double helix structure formed for the CHA reaction was triggered, which immobilized more copper(II) complex on the surface of the working electrode and a strong signal was obtained in the square wave voltammetry (SWV) test. Also, the signal can be re-amplified again in the presence of ferric ion in the electrolyte. Under the optimal conditions, based on these strategies, a sensitive electrochemical biosensor was constructed for the detection of miRNAs in the range of 1 fM to 1 nM and the limit of detection was as low as 0.34 fM. Furthermore, the developed biosensor with excellent selectivity and reproducibility tested on human serum samples shows broad application prospects in the early stage of tumor diagnosis.
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高影响因子
MEDICAL SCIENCE MONITOR
·
2020
·
IF 1.918
Xiaolin Pu,Hua Jiang,Wei Li,Lin Xu,Lin Wang,Yongqian Shu
Background The coatomer protein complex subunit beta 2 (COPB2) gene is upregulated and promotes cell proliferation in some cancer cells. This study aimed to investigate the role of microRNA (miRNA) targeting by COPB2 gene expression in human lung adenocarcinoma cell lines, including NCI-H1975 cells. Material/Methods COPB2 expression in normal human bronchial epithelial cells and lung adenocarcinoma cells was measured by quantitative reverse transcription-polymerase chain reaction (qRT-PCR) and Western blot. NCI-H1975 human lung adenocarcinoma cells were transfected with short-interfering COPB2 (siCOPB2). Cell apoptosis and cell proliferation were evaluated by flow cytometry and Cell Counting Kit-8 (CCK-8) assays, respectively. The transwell assay evaluated cell migration. Targeting of miR-335-3p by COPB2 was predicted using TargetScan 7.2 and verified using a dual-luciferase reporter assay in NCI-H1975 cells. MiR-335-3p mimics were transfected into NCI-H1975 cells. The further functional analysis included detection of protein expression for cyclin D1, tissue inhibitor matrix metalloproteinase-1 (TIMP-1), matrix metallopeptidase 9 (MMP9), Bcl-2, and Bax, to verify the role of miR-335-3p targeting by COPB2 in lung adenocarcinoma cells. Results COPB2 was upregulated in lung adenocarcinoma cells and was a direct target of miR-335-3p mimics. COPB2 knockdown promoted cell apoptosis, inhibited cell migration and proliferation in NCI-H1975 cells. The effects of COPB2 knockdown on NCI-H1975 cells were increased by miR-335-3p mimics, which also further reduced the expression levels of cyclin D1, MMP9, and Bcl-2 and further increased TIMP-1 and Bax by siCOPB2. Conclusions This study showed that COPB2 was the functional target of miR-335-3p in NCI-H1975 human adenocarcinoma cells.
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INTERNATIONAL JOURNAL OF MOLECULAR MEDICINE
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2016
·Vol. 38
·期 6
·1871-1878
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IF 2.348
Xusheng Yuan,Junjun Chen,Min Dai
Paeonol (2'-hydroxy-4'-methoxyacetophenone) is an active component isolated from the root of Paeonia Suffruticosa Andrews. We previously found that paeonol inhibited vascular cell adhesion molecule-1 (VCAM‑1) expression, and thus may be useful for the prevention and treatment of rabbit atherosclerosis (AS); however, the underlying mechanisms are not yet well known. Recently, microRNAs (miRNAs or miRs) have been reported to play an important role in the pathogenesis of AS. In the present study, we examined the effects of paeonol on miRNA-126 (miR‑126) expression, and its ability to inhibit monocyte adhesion to oxidized low-density lipoprotein (ox-LDL)-injured vascular endothelial cells (VECs). VECs were isolated from the rat thoracic aorta and stimulated with ox-LDL (20 mg/l) in the presence of paeonol. We found that miR‑126 had a lower expression in the ox-LDL-injured VECs, and VCAM‑1 was identified as a target gene of miR‑126. Furthermore, paeonol promoted miR‑126 expression and suppressed VCAM‑1 expression at the mRNA and protein level. It also inhibited monocyte adhesion to ox-LDL-injured VECs through the promotion of miR‑126 expression. Furthermore, it was demonstrated that paeonol blocked the activation of the phosphatidylinositol 3-kinase (PI3K)/protein kinase B (Akt)/nuclear factor-κB (NF-κB) signaling pathway by promoting miR-126 expression. Taken together, and to the best of our knowledge, the findings of this study provide the first evidence that paeonol promotes miR‑126 expression to inhibit monocyte adhesion to ox-LDL-injured VECs and block the activation of the PI3K/Akt/NF-κB signaling pathway. Our data suggest that miR‑126 plays a crucial role in vascular inflammation and may be an important therapeutic target in the treatment of AS with the use of paeonol.
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SENSORS AND ACTUATORS B-CHEMICAL
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2022
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IF 9.221
Yu-Yang Wei,Shao-Hua Wu,Qi-Meng Wang,Jian-Jun Sun
A novel split-type and signal-on photoelectrochemical (PEC) biosensor was proposed based on double-strand specific nuclease (DSN)-assisted recycling amplification coupled with photodeposited Ag enhanced BiVO 4 photoanode for sensitive detection of microRNA (miRNA)− 155. In the presence of target miRNA, the capture DNA (cDNA) modified with silver nanocube（AgNC）would hybridize with the miRNA, then DSN-assisted cycle amplification was triggered, leading to the release of AgNCs. The released AgNCs were acidolyzed and photodeposited on the BiVO 4 electrode. Compared with the BiVO 4 electrode, an enhanced photocurrent could be achieved at this obtained BiVO 4 /Ag electrode. And the morphology change from worm-like nanoparticles to nanosheets after Ag photodeposition was observed for the first time. This PEC signal enhancement was mainly contributed to the synergistic action of localized surface plasmon resonance (LSPR) effect of Ag, Schottky junction formed between BiVO 4 and Ag, and the formation of BiVO 4 nanosheets with larger specific surface area. The PEC signal enhancement was positively related to the miRNA-155 concentration, so this method could be used to sensitively and selectively detect miRNA-155. A limit of detection of 8 aM could be achieved. This strategy does not require layer-by-layer modification of the electrode, so it is easy to operate, and owns good stability.
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ANALYTICA CHIMICA ACTA
·
2022
·
IF 6.911
Yang-Yang Hou,Wan-Zhen Xie,Xuecai Tan,Ke-Jing Huang,Jing Xu
A highly sensitivity self-powered biosensor is developed based on T7 exonuclease (T7 Exo) and 3D DNA walker induced rolling circle amplification (RCA) for electrochemical/colorimetric dual-mode detection of microRNA-21 (miRNA-21) with improved reliability. Taking its advantage of fascinating properties, such as high structure defects and good conductivity , graphdiyne is prepared and used to prepare high-performance enzyme biofuel cell. T7 Exo-assisted 3D DNA walker target recognition triggers RCA reaction to obtain a significantly amplified signal response. A capacitor is integrated to the enzyme biofuel cell to further amplify the electrochemical output signal of the self-powered biosensor. In detection system, glucose oxidase catalyzes glucose oxidation to produce hydrogen peroxide, and 3,3′,5,5′-tetramethylbenzidine (TMB) is then catalyzed to generate colored products, so as to achieve the colorimetric detection of the target. Analysis signals of diverse modes are recorded independently. Consequently, detection of microRNA with improved reliability and wider signal response range are achieved by electrochemical/colorimetric dual-mode with detection limits of 0.15 and 33 fM ( S/N = 3) respectively. In addition, the proposed self-powered biosensor successfully applied for the detection of miRNA-21 in human serum samples, confirming its practical applicability in clinical diagnosis. It is powerfully anticipated the proposed self-powered biosensor possesses great potential to be applied to other biomedical domains.
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