MICROCHEMICAL JOURNAL
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2023
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IF 4.8
Mei Li,Qingyou Cai,Yue Meng,Weihong Zheng,Guojiang Mao,Hua Wang
We reported a novel assembled G-wire-induced pyrene excimer switching probe for microRNA (miRNA) amplified detection on the basis of exonuclease III (Exo III)-assisted target recycling and Fe 3 O 4 @PDA nanoparticles. The multifunctional hairpin (HP) probe recognized and effectively bound to target miRNA, which formed a 3′-blunt terminal for Exo-III digestion and miRNA regeneration to initiate another Exo III-mediated cycling cleavage. After digestion, numerous HP-caged c-myc sequences were released, which were positively related to target miRNA and folded to the parallel G-quadruplex with K + . Additionally, Fe 3 O 4 polydopamine nanoparticles (Fe 3 O 4 @PDA NPs)-based magnet separation was conducted to reduce background fluorescence signal, and parallel G-quadruplexes self-asembled into filamentous G-wires with Mg 2+ . G-wires generated from 5′-terminal pyrene-labeled c-myc sequences made close distance between two labeled pyrene molecules, and induced the “turn-on” fluorescence signal of pyrene excimer switching. Moreover, gamma-cyclodextrin (γ-CD) was added, which promoted pyrene moiety, causing signal amplification in pyrene excimer fluorescence. Therefore, miRNA was efficiently measured within the homogeneous solution. This method detected miRNA within the dynamic range of 1 pM to 5 nM, with the miR-141 limit of detection (LOD) being 99 fM. It distinguished perfect-matched and mismatched target miRNAs, and showed an accurate detection of miRNA levels in different cell lines and favorable reproducibility when analyzing miRNAs within spiked serum samples. We propose a promising method for G-wire-based biosensing in disease-related biomarkers detection.
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JOURNAL OF ELECTROANALYTICAL CHEMISTRY
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2020
·
IF 3.807
Xiao Yang,Minghui Feng,Jianfei Xia,Feifei Zhang,Zonghua Wang
A novel electrochemical biosensor based on AuNPs/Ti 3 C 2 MXene three-dimensional nanocomposite for sensitive miRNA-155 detection by exonuclease III (Exo III)-aided cascade target recycling is reported. Ti 3 C 2 MXene is synthesized and used as a support to immobilize Au nanoparticles (AuNPs) due to its special layered structure and metallic properties. The three-dimensional (3D) structure of the AuNPs/Ti 3 C 2 MXene nanocomposite for the biosensor platform leverages the integrated advantages of a large specific surface area, excellent electrical conductivity and electrocatalytic properties . AuNPs are used to immobilize capture DNA (C-DNA) by Au S chemical bonds. The C-DNA was modified with methylene blue (MB) at the 3′ end, resulting in an initial DPV electrochemical signal ( I d ). In the presence of target, miRNA-155 and C-DNA formed a double-stranded structure by complementary base pairing. Subsequently, Exo III digested the 3′ end of the C-DNA in the double-stranded structure and caused the electrochemical signal to “turn off” (the final signal denoted as I h ). By Exo III-aided cascade cleavage of the target, one miRNA-155 chain causes the release of several C-DNAs under certain conditions, which significantly amplifies the peak current difference on the fabricated biosensor (Δ I = I d ﹣ I h ). The electrochemical sensor achieved a detection limit of 0.35 fM (S/N = 3) with a linear range from 1.0 fM to 10 nM. This sensor also shows desirable stability, reproducibility and specificity.
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MICROCHIMICA ACTA
·
2020
·Vol. 187
·期 4
·1-9
·
IF 6.232
Ji Xiaoting,Wang Zhenbo,Niu Shuyan,Ding Caifeng
The present study shows that a dual-signal nanoprobe consisting of DNAzyme-functionalized porous carbon nanospheres (PCNs) responds to microRNA-21 and zinc ion (Zn 2+ ). The fluorescent probe undergoes an increase in the fluorescence intensity of fluorescein isothiocyanate (FITC) (with excitation/emission wavelengths at 488/517 nm) and the fluorescence intensity of cyanine-5 (Cy5) (with excitation/emission wavelengths at 633/670 nm) in the presence of microRNA-21 and Zn 2+ . The recognition between microRNA-21 and its complementary strand in the PCNs induces the separation of Zn 2+ -specific DNAzyme from PCNs, thus resulting in the increase of green fluorescence, and the exogenous Zn 2+ triggers the rupture of cleavage strand of DNAzyme and recovery of red fluorescence. This nanoprobe allows us to acquire in vitro the determination of microRNA-21 in the range of 2–300 nM with a detection limit of 0.57 nM and the determination of Zn 2+ in the range 2–100 nM with a detection limit of 0.43 nM, and in situ simultaneous imaging in MCF-7 breast cancer cells. Therefore, this strategy permits to obtain the expression levels of different biomarkers in living cells, providing a useful tool for diagnosis of cancers and understanding their biological process.
Schematic representation of the DNAzyme-functionalized porous carbon nanospheres for the imaging analysis of microRNA-21 and Zn2+ in living cells.
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MICROCHIMICA ACTA
·
2021
·Vol. 188
·期 8
·1-9
·
IF 5.833
Yang Jiao,Fu Shilan,Luo Fang,Guo Longhua,Qiu Bin,Lin Zhenyu
MicroRNAs can serve as biomarkers for many cancers, so it is significant to develop simple and sensitive strategies for microRNAs detection. Photoelectrochemical (PEC) detection has the advantages of simple equipment and high sensitivity. But in conventional PEC DNA sensors, tedious immobilization procedures of photoactive materials and capture probes on electrode surfaces are inevitable. To overcome those limitations, a homogeneous PEC biosensor based on target-responsive hydrogels has been developed (miRNA-155 has been chosen as a model target). PEC signal molecules (TiO 2 nanoparticles, TiO 2 NPs) were embedded in DNA hydrogels formed by hyaluronic acid sodium salt, amine-modified DNA double strands, and polyethylenimine rich in amine groups. In the presence of the target, DNA double strands in hydrogel were nicked by endonuclease and TiO 2 NPs were released to the supernate and a high PEC response was obtained when collecting the supernate for PEC test, while almost no TiO 2 NPs released in the absence of the target. Thanks to the exonuclease III and nicking endonuclease Nb.BbvCI-assisted cascaded amplification strategy, the proposed biosensor exhibits high sensitivity toward miRNA-155 with a low detection limit of 0.41 fM and a wide linear range from 1.0 fM to 100 pM. Since this method circumvents tedious electrode modification procedures, the proposed technique exhibits the advantages of simplicity and good reproducibility. Moreover, the prepared hydrogels have outstanding storage stability, so that they can be prepared in advance and shorten detection time. This biosensing platform provides a versatile strategy for the construction of homogeneous PEC biosensors for the detection of diverse targets. Graphical abstract
Photoelectrochemical detection techniques have been coupled with controlled release system to develop an immobilization-free microRNA biosensor. High sensitivity has been realized based on cascaded signal amplification strategy, and the proposed biosensor has been applied to detect the target in real sample with satisfied results. Since no tedious electrode modifications, the proposed homogeneous PEC sensor exhibits high reproducibility and good stability.
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ANALYTICAL CHEMISTRY
·
2019
·
IF 6.35
Jing Ye,Guoyong Liu,Mengxia Yan,Qiuju Zhu,Liping Zhu,Jianshe Huang,Xiurong Yang
Inspired by the coreactive activity of carbon nanodots (CDs) and branched polyethylenimine (BPEI) toward electrochemiluminescence (ECL) of Ru(bpy)32+, a highly luminescent and self-enhanced ECL nanohybrid (Ru-BCDs) was synthesized through covalently linking BPEI-coated carbon dots (BCDs) with Tris (4,4′-dicarboxylic acid-2,2′-bipyridyl) ruthenium(II) dichloride (Ru(dcbpy)32+). The composition and morphological characterization demonstrated that the spherical Ru-BCDs particles with 12.1 ± 1.4 nm diameter were obtained. The enhanced ECL property of Ru-BCDs was proved to originate from the dual coreactive contribution of BPEI and CDs as coreactants as well as the intramolecular electron transfer process, which could shorten the electron transfer path and minimize energy loss. A carbon nitride nanosheet (CNN) was utilized to stabilize the Ru-BCDs-modified glassy carbon electrode, which greatly improved the stability of solid-state ECL. By utilizing the affinity discrepancy of the CNN to single-stranded and double-stranded nucleic acids, a label-free and signal-on ECL biosensor was constructed for the determination of microRNA-133a (miR-133a), a potential biomarker of acute myocardial infarction. The designed biosensor exhibited good performance of miR-133a detection with a detection limit of 60 fM and could be used for the detection of real human serum with satisfactory results. The self-enhanced ECL nanohybrid with distinguished ECL efficiency holds a promising prospect in biosensing and bioimaging applications.
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JOURNAL OF MOLECULAR NEUROSCIENCE
·
2019
·Vol. 68
·期 2
·181-190
·
IF 2.577
Yan Penghui,Wu Xuejian,Liu Xiaokang,Cai Yingchun,Shao Chenglong,Zhu Guangduo
Microglial activation and inflammatory response played an important role in the secondary injury of spinal cord injury (SCI). Several microRNAs were associated with this procedure, but the underlying molecular mechanism was poorly understood. Sprague-Dawley (SD) rats were divided into four groups: SCI group ( n = 7), agomiR-325-3p group ( n = 7), and their control groups. Expression of miR-325-3p and proteins in epidermal growth factor receptor (EGFR)/mitogen-activated protein kinase (MAPK) signaling pathway was evaluated in microglia from SCI rats and primary microglia/BV2 cells activated by lipopolysaccharide (LPS). Concentrations of interleukin-1β (IL-1β) and tumor necrosis factor α (TNF-α) in supernatants were measured by ELISA. Low expression of miR-325-3p and activation of EGFR/MAPK was observed in microglia of SCI and LPS-induced primary microglia. Overexpression of miR-325-3p in LPS-induced BV2 cells inhibited microglial activation and release of TNF-α and IL-1β. Luciferase reporter assay confirmed that miR-325-3p negatively regulated EGFR by targeting its 3′-untranslated regions. Additionally, agomiR-325-3p inhibited the activation of microglia and EGFR/MAPK, alleviating the inflammatory response. These results indicated that miR-325-3p attenuated secondary injury after SCI through inhibition of EGFR/MAPK signaling pathway, the microglial activation, and the release of inflammatory cytokines, suggesting that miR-325-3p may be employed as a therapeutic target for SCI.
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INTERNATIONAL JOURNAL OF BIOLOGICAL MACROMOLECULES
·
2025
·
IF 8.5
Xiaomei Mu,Shixiu Xiao,Shulin Zhao,Jianniao Tian
In this work, a fast signal amplification system mediated by self-replicating catalytic hairpin self-assembly (SCHA) was established for microRNA-155 using near-infrared DNA-Ag Nanoclusters (DNA-AgNCs) as fluorescence signal output. Among them, two fission target-like DNA sequences are merged into two hairpin DNA H1 and H2, and the AgNCs template sequence is designed at the sticky end of H1 and H2. The target can be recycled in the system to form a double-stranded DNA structure (H1–H2), which will detach the H1/H2-AgNCs from the surface of the polypyrrole nanoparticles (PPy NPs) and cause the near-infrared fluorescence signal of DNA-AgNCs to be restored. At this point, the two-split target-like DNA sequences will be reassembled to initiator DNA. The acquired replicas can also be recycled as a brand-new activation unit to initiate the SCHA response, resulting in rapid replication of the target/triggered DNA, accompanied by the generation of higher fluorescence signals. This autocatalytic signal amplification approach has been successfully applicable to fast signal amplification, enzyme-free and label-free for microRNA-155 assay in biological samples, and the detection limit (LOD) is 240 fM (S/N = 3). At the same time, this SCHA system can realize intracellular microRNA fluorescence imaging, which presents a promising approach to developing advanced molecular diagnostic tools.
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JOURNAL OF CELLULAR AND MOLECULAR MEDICINE
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2021
·Vol. 25
·期 8
·3744-3753
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IF 5.31
Zhong-Sheng Xiao,Lei Zhao,Xiao-Ning Zhang,Han-Xian Li,Zhi-Hui Yin
It has been reported that rs67085638 in long non-coding RNAs (lncRNA)-CCAT1 was associated with the risk of tumorigenesis. Also, CCAT1 could affect chemoresistance of cancer cells to paclitaxel (PTX) via regulating miR-24-3p and FSCN1 expression. In this study, we aimed to investigate the effect of rs67085638 on the expression of CCAT1/miR-24-3p/FSCN1 and the response of colon cancer to the treatment of PTX. 48 colon cancer patients were recruited and grouped by their genotypes of rs67085638 polymorphism as a CC group (N = 28) and a CT group (N = 20). PCR analysis, IHC assay and Western blot, TUNEL assay and flow cytometry were conducted. LncRNA-CCAT1 and FSCN1 mRNA/protein were overexpressed, whereas miR-24-3p was down-regulated in the CT-genotyped patients and cells compared with those in the CC-genotyped patients and cells. The survival of colon cancer cells was decreased, whereas the apoptosis of colon cancer cells was increased by PTX treatment in a dose-dependent manner. MiR-24-3p was validated to target lncRNA-CCAT1 and FSCN1 mRNA, and the overexpression of CCAT1 could reduce the expression of miR-24-3p although elevating the expression of FSCN1. Knockdown of lncRNA-CCAT1 partly reversed the suppressed growth of CT-genotyped tumours. And the knockdown of lncRNA-CCAT1 partly reversed the dysregulation of lncRNA-CCAT1 and FSCN1 mRNA/protein in rs67085638-CT + NC shRNA mice. The findings of this study demonstrated that the presence of the minor allele of rs67085638 increased the expression of CCAT1 and accordingly enhanced the resistance to PTX. Down-regulation of CCAT1 significantly re-stored the sensitivity to PTX of colon cancer cells.
研究领域:
癌症生物学
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分子生物学
·
遗传学
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Molecular Therapy-Nucleic Acids
·
2023
·
IF 8.8
Chen Jun Guo,Xiu Li Cao,Yu Fei Zhang,Kang Yi Yue,Jing Han,Hong Yan,Hua Han,Min Hua Zheng
Inherited and age-related retinal degenerations are the commonest causes of blindness without effective treatments. Retinal progenitor cells (RPCs), which have the multipotency to differentiate into various retinal cell types, are regarded as a promising source of cell transplantation therapy for retinal degenerative diseases. However, the self-limited expansion of RPCs causes difficulty in cell source supply and restrict its clinical treatment. In this work, we found that inhibition of microRNA-449a (miR-449a) in RPCs can promote proliferation and inhibit apoptosis of RPCs, partially through upregulating Notch signaling. Further optimization of transduction miR-449a inhibitor into RPCs by endothelial cell-derived exosomes can promote the survival of RPCs transplanted in vivo and reduce cell apoptosis in retinal degeneration mouse models. In summary, these studies have shown that exosome-miR-449a inhibitor can effectively promote the expansion of RPCs in vitro and enhance transplanted RPCs survival in vivo , which might provide a novel intervention strategy for retinal degenerations in the future.
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Evidence-based Complementary and Alternative Medicine
·
2023
·
IF 2.65
Hui Wang,HaiTang Ren
The action mechanism in which circular RNA (circ) SMARCA5 targeted nasopharyngeal carcinoma (NPC) cell proliferation, migration, invasion, and apoptosis via microRNA (miR)-582-3p/phosphatase and tensin homolog (PTEN) axis was explored. The examination was performed via reverse transcription-quantitative polymerase chain reaction (RT-qPCR), discovering that circSMARCA5 was elevated while miR-582-3p was silenced in NPC tissues and cells. E-cadherin and N-cadherin were detected. The results illustrated transfection with si-circSMARCA5 or miR-582-3p-mimic was available to repress cancer cell advancement, and E-cadherin was augmented. Transfection with pcDNA 3.1-circSMARCA5 or miR-582-3p-inhibitor was available to accelerate cancer cell advancement, and N-cadherin was augmented. MiR-582-3p-inhibitor blocked the suppression of si-circSMARCA5 on NPC. The si-PTEN blocked the malignant behavior of pcDNA 3.1-circSMARCA5 against NPC. The binding sites between circSMARCA5 and miR-582-3p and between miR-582-3p and PTEN were verified. Linear analysis results illuminated the expression pattern of circSMARCA5 was opposite to miR-582-3p, while the expression pattern of circSMARCA5 was positively associated with PTEN. In brief, the results of the research clarified circSMARCA5 modulated NPC cells’ vital movement via the miR-582-3p/PTEN molecular axis.
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Drug Design Development and Therapy
·
2022
·
IF 4.319
Shurong Duan,Fei Wang,Jingwei Cao,Chunyan Wang
Introduction Intracerebral hemorrhage (ICH) is a devastating type of stroke with high mortality, and the effective therapies for ICH remain to be explored. Exosomes (Exos) have been found to play important roles in cell communication by transferring molecules, including microRNAs (miRNAs/miRs). MiRNAs are critical regulators of genes involved in many various biological processes and have been demonstrated to aggravate or alleviate brain damages induced by ICH. The aim of the present study was to investigate the effect of Exos derived from miR-146a-5p-enriched bone marrow mesenchymal stem cells (BMSCs-miR-146a-5p-Exos) on experimental ICH.Methods ICH was induced in adult male Sprague-Dawley rats by an intrastriatal injection of collagenase type IV. At 24 h after surgery, Exos were administrated. For detecting apoptotic cells, TUNEL staining was performed using an in situ Cell Death Detection Kit. Fluoro-Jade B staining was performed to detect degenerating neurons. Immunofluorescence assay was performed to detect the expression of myeloperoxidase (MPO) and OX-42. The binding of miR-146a-5p and its target genes was confirmed by luciferase reporter assay.Results At 24 h after surgery, BMSCs-miR-146a-5p-Exos administration significantly improved neurological function, reduced apoptotic and degenerative neurons, and inhibited inflammatory response. Furthermore, miR-146a-5p-enriched Exos obviously inhibited the M1 polarization of microglia after ICH in rats, accompanied by the reduced expression of pro-inflammatory mediators releasing by M1 microglia including inducible nitric oxide synthase (iNOS), cyclooxygenase-2 (COX-2) and monocyte chemoattractant protein-1 (MCP-1). Finally, we observed that miR-146a-5p directly targeted interleukin-1 receptor-associated kinase1 (IRAK1) and nuclear factor of activated T cells 5 (NFAT5), which contributed to the inflammation response and the polarization of M1 microglia/macrophages.Conclusion We demonstrated that miR-146a-5p-riched BMSCs-Exos could offer neuroprotection and functional improvements after ICH through reducing neuronal apoptosis, and inflammation associated with the inhibition of microglial M1 polarization by downregulating the expression of IRAK1 and NFAT5.
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JOURNAL OF EXPERIMENTAL & CLINICAL CANCER RESEARCH
·
2022
·Vol. 41
·期 1
·1-15
·
IF 12.658
Cao Yi,Tu Yi,Xiong Jianbo,Tan Shengxing,Luo Lianghua,Wu Ahao,Shu Xufeng,Jie Zhigang,Li Zhengrong
Background Extracellular vesicles (EVs) derived from tumor-associated macrophages are implicated in the progression and metastasis of gastric cancer (GC) via the transfer of molecular cargo RNAs. We aimed to decipher the impact of microRNA (miR)-15b-5p transferred by M2 macrophage-derived EVs in the metastasis of GC.Methods Expression of miR-15b-5p was assessed and the downstream genes of miR-15b-5p were analyzed. GC cells were subjected to gain- and loss-of function experiments for miR-15b-5p, BRMS1, and DAPK1. M2 macrophage-derived EVs were extracted, identified, and subjected to co-culture with GC cells and their biological behaviors were analyzed. A lung metastasis model in nude mice was established to determine the effects of miR-15b-5p on tumor metastasis in vivo.Results miR-15b-5p was upregulated in GC tissues and cells as well as in M2 macrophage-derived EVs. miR-15b-5p promoted the proliferative and invasive potentials, and epithelial-mesenchymal transition (EMT) of GC cells. M2 macrophage-derived EVs could transfer miR-15b-5p into GC cells where it targeted BRMS1 by binding to its 3’UTR. BRMS1 was enriched in the DAPK1 promoter region and promoted its transcription, thereby arresting the proliferative and invasive potentials, and EMT of GC cells. In vivo experiments demonstrated that orthotopic implantation of miR-15b-5p overexpressing GC cells in nude mice displayed led to enhanced tumor metastasis by inhibiting the BRMS1/DAPK1 axis.Conclusion sOverall, miR-15b-5p delivered by M2 macrophage-derived EVs constitutes a molecular mechanism implicated in the metastasis of GC, and may thus be considered as a novel therapeutic target for its treatment.
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