Journal of Inflammation Research
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2023
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IF 4.5
Dongsheng Zhu,Zhitao Zhu,Han Qi
Objective Long non-coding RNA plays an important role in osteogenic differentiation. Nuclear enriched abundant transcript 1 (NEAT1) has been revealed to promote osteogenic differentiation in human bone marrow mesenchymal stem cells (hBMSCs), but the underlying regulatory mechanism remains unknown in acute suppurative osteomyelitis of children.Methods Osteogenic medium (OM) was used to induce osteogenic differentiation. Quantitative real-time PCR and Western blotting were used to evaluate gene expression. The effects of NEAT1, microRNA 339–5p (miR-339-5p), and salmonella pathogenicity island 1 (SPI1) on osteogenic differentiation were assessed in vitro using alizarin red S staining assays and alkaline phosphatase activity. Interactions between NEAT1, miR-339-5p, and SPI1 were identified using immunoprecipitation, luciferase reporter assays, and chromatin immunoprecipitation.Results During osteogenic differentiation, expression of NEAT1 was up-regulated in hBMSCs, and miR-339-5p level was down during osteogenic differentiation. Knockdown of NEAT1 reduced the osteogenic differentiation of hBMSCs, and down-regulation of miR-339-5p may counteract the effect of NEAT1 silencing. SPI1 was a target of miR-339-5p by luciferase reporter assay and was also a transcription factor of NEAT1 by chromatin immunoprecipitation. A positive NEAT1-miR-339-5p-SPI1 feedback loop was found to be present during osteogenic differentiation in hBMSCs.Conclusion It was the first study to reveal that the NEAT1-miR-339-5p-SPI1 feedback loop can promote osteogenic differentiation in hBMSCs and shed a new light on the role of NEAT1 during osteogenic differentiation.
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ANALYTICA CHIMICA ACTA
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2023
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IF 6.2
Yuan Su,Zilian Jiang,Yahui Wang,Hongfang Zhang
The interesting adsorption affinity of two-dimensional nanosheets to single stranded over double stranded nucleic acids have stimulated the exploration of these materials in biosensing. Herein, MoS 2 nanosheets decorated anodic aluminum oxide (AAO) membrane was simply prepared by suction filtration. The MoS 2 /AAO hybrid membrane was initially applied to the electrochemical detection of microRNA using let-7a as the model. When let-7a was incubated with its complementary DNA , double stranded DNA-RNA formed and which displayed weak adsorption capability to the hybrid membrane. And thus the steric effect combining the electrostatic repulsion of the backbone phosphate of nucleic acids for [Fe(CN) 6 ] 3− transport across the hybrid membrane varied with the concentration of let-7a. In this way, a label-free electrochemical detection method for microRNA was established by monitoring the change of the redox current of [Fe(CN) 6 ] 3− . To further improve the detection sensitivity of the method, we proposed two separate strategies focusing on the amplification of the target-induced steric hindrance with DNA nanostructure and the magnification of the electrode sensitivity for [Fe(CN) 6 ] 3− by electrode modification. By using the two strategies, the hybrid membrane based-detection method exhibited broad linear range, low detection limit and good selectivity as well as reproducibility. Therefore, this study provided a proof-of-concept for the application of two-dimensional material to nucleic acids detection.
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Journal of Thoracic Disease
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2023
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IF 2.5
Dianbao Zhang,Yukun Yang,Yixin Kang,Dongjie Xie,Xianfen Zhang,Jiqing Hao
Background An increasing amount of evidence has confirmed that the altered expression of microRNAs (miRNAs) is critical to the mechanism underlying primary and even acquired resistance to tyrosine kinase inhibitors (TKIs). However, studies on the linkage between the altered miRNAs expression and osimertinib resistance are few, and the effect of miRNAs in this context is still unclear. In the light of this, we hypothesized that the differential expression of multiple miRNAs is the driver in the osimertinib resistance process. Thus, the aim of our study was to find differentially expressed miRNAs in non-small cell lung cancer cells resistant to osimertinib. Methods An AZD9291(Osimertinib)-resistant cell line model was constructed, and the differential miRNAs between epidermal growth factor receptor (EGFR)-sensitive cell lines A549 and H1975 and the corresponding drug-resistant cell lines were identified via biosynthesis analysis. Results In the A549 osimertinib-resistant cell line, 93 miRNAs were upregulated and 94 miRNAs were downregulated. In the H1975 osimertinib-resistant cell line, 124 miRNAs were upregulated and 53 miRNAs were downregulated. Finally, 7 significantly different miRNAs were screened using Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analysis. Conclusions This study on the mechanism of target therapy in lung cancer systematically and comprehensively examined the miRNAs involved in osimertinib resistance. It was found that miR-708-5p, miR-708-3p, miR-10395-3p, miR-7704 miR-34a-5p, miR-19b-1-5p, and miR-219a-5p may play key roles in osimertinib resistance.
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ANALYTICAL CHEMISTRY
·
2019
·
IF 6.35
Yang Li,Cheng Zhi Huang,Yuan Fang Li
High sensitivity and accuracy are two key issues that are critical for electrochemiluminescence (ECL) detection, especially for low-abundance nucleic acid detection. However, research on the construction of biosensors has mainly been through a step-by-step approach, which will increase the systematic error and affect the accuracy of the detection. Here we propose a novel strategy of introduction of a branched hybridization chain reaction (bHCR) circuit to a terbium(II) organic gel (TOG) modified electrode in one step to achieve both sensitive detection and simplified modification steps. The sensitivity of the biosensor was elevated by the cascade bHCR circuit that was activated by miRNA-141 and operated like a molecular machine to form hyperbranched DNA nanostructures. Benefiting from molecular programming, the obtained nanostructures carried a large number of dopamine molecules, which can effectively quench the ECL signal of emitters and achieve a low limit of detection (0.18 fM). Impressively, the proposed one-step approach was almost the easiest way to modify nucleic acids to electrodes. In this way, the introduction of a high-molecular-weight DNA structure in one step avoided the errors that may result from the stepwise modification of low-molecular-weight nucleic sequences into the electrode. Considering the accessible operation, favorable performance, and high universality of this strategy, this work may be used to analyze other microRNAs and further clinical diagnosis.
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TALANTA
·
2019
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IF 4.916
Feng Zhang,Shuang Wang,Jian feng,Rong Zou,Ling Xiang,Changqun Cai
A molecular beacons (MBs) loaded on molybdenum disulfide (MoS 2 ) nanosheets as fluorescence probes for sensitive and versatile detection of microRNAs (miRNAs) through hybridization chain reaction (HCR) has been designed. MoS 2 was used as a adsorbent to capture the MBs and a selective fluorescence quencher to reduce the background signal. In the absence of miRNAs, HCR could not be triggered due to the stability of MB probes. The probes attached to the MoS 2 surface, efficiently quenching fluorescence of the G-quadruplex/Thioflavin T . However, the presence of target miRNAs triggers the HCR process to generate large amount of HCR products. Meanwhile, the HCR products of long nanowires chain with abundant G-quadruplexes could not be adsorbed on the surface of MoS 2 , and therefore detach from the MoS 2 . Consequently, Thioflavin T could be embedded in G-quadruplexes and produced strong fluorescence signal. This fluorescence emission signal could achieve detection of miRNA as low as 4.2 pM and a wide linear ranges from 0.1 to 100 nM. In addition, a versatile fluorescence probe has been developed for detection of miRNA-21 by changing the miRNA-recognition domain of MB. Thus, the fluorescent probe would be a potential alternative tool for biomedical research and clinical molecular diagnostics.
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ANALYTICAL CHEMISTRY
·
2019
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IF 6.35
Huan Gao,Junfang Zhang,Ying Liu,Wenwen Tu,Tianxiang Wei,Zhihui Dai
Development of sensitive detection methods for microRNAs has realistic application value for early clinical analysis and accurate diagnosis. In this study, a triple-helix molecular switch electrochemiluminescence resonance energy transfer (ECL-RET) nanoamplifier was designed to construct an electrochemiluminescence (ECL) biosensor for microRNA determination. The newly synthesized S-doped Lu2O3, which shows a 3 times better ECL performance than Lu2O3, was chosen as the donor in this ECL-RET system. Accordingly, Ag2S quantum dots (QDs) were used as the matched acceptor. They exhibited overlapping spectra and efficient energy transfer between each other. Furthermore, using a triple-helix switch structure with an improved quenching effect for signal amplification and a nano-DNA walker transformational system as a powerful method for target amplification, a supernanoamplifier was achieved. As a consequence, the proposed ECL biosensor for microRNA-141 detection exhibited good analytical performance with a low detection limit (16 aM). The sensor not only led to the development of a novel ECL-RET pair but also provided a strong nanoamplifier for biosensing platform construction, which may offer some new considerations regarding material design, signal amplification, and show promising application value in biomedical and clinical diagnosis.
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Frontiers in Cardiovascular Medicine
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2022
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IF 5.846
Hongge Fa,Dandan Xiao,Wenguang Chang,Lin Ding,Lanting Yang,Yu Wang,Mengyu Wang,Jianxun Wang
Objective Many studies have reported that microRNAs (miRs) are involved in the regulation of doxorubicin (DOX)-induced cardiotoxicity. MiR-194-5p has been reported significantly upregulated in patients with myocardial infarction; however, its role in myocardial diseases is still unclear. Various stimuluses can trigger the endoplasmic reticulum (ER) stress and it may activate the apoptosis signals eventually. This study aims to explore the regulatory role of miR-194-5p in DOX-induced ER stress and cardiomyocyte apoptosis. Methods H9c2 was treated with 2 μM DOX to induce apoptosis, which is to stimulate the DOX-induced cardiotoxicity model. The expression of miR-194-5p was detected by quantitative real-time PCR (qRT-PCR); the interaction between miR-194-5p and P21-activated kinase 2 (PAK2) was tested by dual luciferase reporter assay; terminal deoxynucleotidyl transferase dUTP nick-end labeling (TUNEL) assay and caspase-3/7 activity were used to assess apoptosis; trypan blue staining was applied to measure cell death; Western blotting was performed to detect protein expressions; and ER-related factors splicing X-box binding protein 1 (XBP1s) was detected by polyacrylamide gel electrophoresis and immunofluorescence to verify the activation of ER stress. Results MiR-194-5p was upregulated in cardiomyocytes and mouse heart tissue with DOX treatment, while the protein level of PAK2 was downregulated. PAK2 was predicted as the target of miR-194-5p; hence, dual luciferase reporter assay indicated that miR-194-5p directly interacted with PAK2 and inhibited its expression. TUNEL assay, caspase-3/7 activity test, and trypan blue stain results showed that either inhibition of miR-194-5p or overexpression of PAK2 reduced DOX-induced cardiomyocyte apoptosis. Silencing of miR-194-5p also improved DOX-induced cardiac dysfunction. In addition, DOX could induce ER stress in H9c2, which led to XBP1 and caspase-12 activation. The expression level of XBP1s with DOX treatment increased first then decreased. Overexpression of XBP1s suppressed DOX-induced caspase-3/7 activity elevation as well as the expression of cleaved caspase-12, which protected cardiomyocyte from apoptosis. Additionally, the activation of XBP1s was regulated by miR-194-5p and PAK2. Conclusion Our findings revealed that silencing miR-194-5p could alleviate DOX-induced cardiotoxicity via PAK2 and XBP1s in vitro and in vivo . Thus, the novel miR-194-5p/PAK2/XBP1s axis might be the potential prevention/treatment targets for cancer patients receiving DOX treatment.
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LANGMUIR
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2022
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IF 4.331
Fa-Ting Xie,Yu-Long Li,Tong Yang,Yun-Hui Yang,Rong Hu
Electrochemical nanotags with strong signal input are necessary for a ratiometric electrochemical sensor to overcome the drawbacks of inaccurate detection results. In this paper, the metal–organic framework (MOF) UiO-66 was utilized as an electrochemical signal tag. A stable and strong current response at +0.9 V can be detected in neutral conditions. MicroRNA (miRNA) was employed as the model analyte. Herein, an enzyme-free DNA-walker-based ultrasensitive ratiometric electrochemical biosensor in combination with Zr MOF (UiO-66) signal tags to detect miRNA was demonstrated. In the presence of miRNA, the autonomous walker movement can be initiated by miRNA, leading to the release of biotin-modified fragments. Thus, streptavidin-labeled UiO-66 nanomaterials were not bound to the electrode, generating a low signal response of UiO-66 at +0.9 V. However, the current signal of electrolyte solution as reference at +0.2 V was increased due to the enhancement of electrode conductivity. This ratiometic sensor demonstrated high sensitivity, selectivity, and reproducibility. It can eliminate the disturbance of environmental factors and basic electrode characteristics, providing more accurate signals. A limit of detection (LOD) of 0.17 fM was achieved. Moreover, the method was also used to detect miRNA-21 spiked in real serum samples.
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ELECTROCHIMICA ACTA
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2022
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IF 7.336
Qianqing Wu,Zhenhui Li,Qianwei Liang,Rongkai Ye,Shuzhou Guo,Xiaobing Zeng,Jianqiang Hu,Aiqing Li
MicroRNAs (miRNAs) are considered as potential biomarkers for early diagnosis and prognostic assessment of diabetic nephropathy. In this work, an electrochemical biosensor for ultrasensitive detection of miRNA-377 was constructed based on MXene-Au nanocomposites and G-quadruplex nano-amplification strategy. As a promising nanocarrier, taking advantage of leveraging synergistic effects between Au nanoparticles (AuNPs) and MXene nanosheet, MXene-Au nanocomposites exhibited excellent electronic conductivity and provided massive active sites for DNA capture probe immobilization by Au-S bonds. AuNPs modified with Guanine-rich sequence DNA detection probes were designed as signal amplification nano-labels. Specifically, by inducing the transition of Guanine-rich detection probes to G-quadruplex, the strong affinity interaction between methylene blue and G-quadruplex could not only reflect trace concentration information of miRNA-377, but also lead to the further enhancement of electrochemical signal (2.7-fold). As a result, this newly designed biosensor exhibited superior sensing performance with a wide linear range from 10 aM to 100 pM and the limit of detection was as low as 1.35 aM. Compared with biosensors based on other nanocomposites and reported miRNA-377 biosensors, the proposed sensing platform did not require any thermal cycling or reverse transcription process, meeting the miRNA sensing requirements of convenient, sensitive, specific and stable. Furthermore, the as-constructed biosensor also displayed good selectivity, which was applied to accurate detection of miRNA-377 in human serum samples with satisfactory sensitivity, suggesting the biosensor system has promising applications in biological researches and early clinical diagnosis for diabetic nephropathy.
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ANALYST
·
2020
·Vol. 146
·期 6
·1924-1931
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IF 3.978
Zibo Gao,Huijie Yuan,Yanhua Mao,Lihua Ding,Clement Yaw Effah,Sitian He,Leiliang He,Li-e Liu,Songcheng Yu,Yilin Wang,Jia Wang,Yongmei Tian,Fei Yu,Hongchao Guo,Lijun Miao,Lingbo Qu,Yongjun Wu
MicroRNAs (miRNAs) encapsulated in tumor-derived exosomes are becoming ideal biomarkers for the early diagnosis and prognosis of lung cancer. However, the accuracy and sensitivity are often hampered by the extraction process of exosomal miRNA using traditional methods. Herein, this study developed a fluorogenic quantitative detection method for exosomal miRNA using the fluorescence quenching properties of molybdenum disulfide (MoS2) nanosheets and the enzyme-assisted signal amplification properties of duplex-specific nuclease (DSN). First, a fluorescently-labeled nucleic acid probe was used to hybridize the target miRNA to form a DNA/RNA hybrid structure. Under the action of the DSN, the DNA single strand in the DNA/RNA hybrid strand was selectively digested into smaller oligonucleotide fragments. At the same time, the released miRNA target triggers the next reaction cycle, so as to achieve signal amplification. Then, MoS2 was used to selectively quench the fluorescence of the undigested probe leaving the fluorescent signal of the fluorescently-labeled probe fragments. The fluorometric signals for miRNA-21 had a maximum excitation/emission wavelength of 488/518 nm. Most importantly, the biosensor was then applied for the accurate quantitative detection of miRNA-21 in exosome lysates extracted from human plasma and this method was able to successfully distinguish lung cancer patients from healthy people. This biosensor provides a simple, rapid, and a highly specific quantitative method for exosomal miRNA and has promising potential to be used in the early diagnosis of lung cancer.
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LUMINESCENCE
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2020
·Vol. 35
·期 6
·835-844
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IF 1.855
Zheng Chang,Jing Feng,Xingwang Zheng
In this paper, a convenient reverse-phase microemulsion method for the synthesis of SiO 2 nanoparticles (NPs) by simply introducing the chitosan and fluorescent dye of lucigenin during the formation reaction of SiO 2 NPs was proposed. Addition of chitosan can make the SiO 2 NPs porous, and increases lucigenin molecule incorporation into chitosan/SiO 2 NPs nanopores based on electrostatic interaction and supermolecular forces. Therefore, fluorescence quantum yield of the lucigenin/chitosan/SiO 2 composite nanoparticles was increased by introduction of chitosan and compared with lucigenin/SiO 2 NPs without chitosan. Because the number of negative charges carried when using single-stranded DNA (ssDNA) was different from that of double-stranded DNA (dsDNA), the numbers of lucigenin/chitosan/SiO 2 composite nanoparticles with positive charge adsorbed using ssDNA or dsDNA were different. Consequently, fluorescence intensity caused using ssDNA or dsDNA/miRNA was clearly discriminative. With increase in target DNA/miRNA concentration, the difference in fluorescence intensity also increased, resulting in a good linear relationship between fluorescence intensity sensitizing value and target miRNA concentrations. Therefore, a new fluorescence analysis method for direct detection of let-7a in human gastric cancer cell samples without enzyme, label free and no immobilization was established using lucigenin/chitosan/SiO 2 composite nanoparticles as a DNA hybrid indicator. The proposed method had high sensitivity and selectivity, low cost and the detection limit was 10 fM (S/ N = 3).
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ACS Sensors
·
2019
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IF 6.944
Lin Shi,Yudie Sun,Lan Mi,Tao Li
Portable chemiluminescence (CL) imaging with a smartphone has shown a great promise for point-of-care testing of diseases, especially for acute myocardial infarction (AMI), which may occur abruptly. A challenge remains how to improve the imaging sensitivity that usually is several orders of magnitude lower than those of counterpart methodologies using the sophisticated equipment. Toward this goal, here, we report the target-triggered in situ growth of AuNP@hairpin-DNA nanoprobes into spherical nucleic acid enzymes (SNAzymes), which serve as both nanolabels and amplifiers for portable CL imaging of microRNAs (miRNAs) with an ultrahigh sensitivity comparable to that of the instrumental measurement under same conditions. A G-quadruplex (G4) DNA dense layer is dynamically produced on the gold nanocore via a DNAzyme machine-driven hairpin cleaving and captures the cofactor hemin to form the SNAzymes with higher peroxidase activity and stronger nuclease resistance than the commonly used G4 DNAzymes. The matured SNAzymes are then utilized as catalytic labels in a luminol-artesunate CL system for miRNA imaging with a smartphone as the portable detector. In this way, two AMI-related miRNAs, miRNA-499 and miRNA-133a, are successfully detected in real patients’ serum with a naked eye-visualized CL change at 10 fM, showing a 5 order of magnitude improvement on the sensitivity of visualizing the same disease markers in clinical circulating blood as compared to the reported strategy. In addition, a good selectivity of our developed CL imaging platform is demonstrated. These unique features make it promising to employ this portable imaging platform for clinical AMI diagnosis.
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