PeerJ
·
2021
·
IF 2.984
Wenwen Wang,Wenwen Zhang,Yuanjing Hu
Background Chemotherapy resistance, especially platinum resistance, is the main cause of poor prognosis of ovarian cancer. It is of great urgency to find molecular markers and mechanism related to platinum resistance in ovarian cancer. Methods One mRNA dataset ( GSE28739 ) and one miRNA dataset ( GSE25202 ) were acquired from Gene Expression Omnibus (GEO) database. The GEO2R tool was used to screen out differentially expressed genes (DEGs) and differentially expressed miRNAs (DE-miRNAs) between platinum-resistant and platinum-sensitive ovarian cancer patients. Gene Ontology (GO) function and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analysis for DEGs were performed using the DAVID to present the most visibly enriched pathways. Protein–protein interaction (PPI) of these DEGs was constructed based on the information of the STRING database. Hub genes related to platinum resistance were visualized by Cytoscape software. Then, we chose seven interested hub genes to further validate using qRT-PCR in A2780 ovarian cancer cell lines. And, at last, the TF-miRNA-target genes regulatory network was predicted and constructed using miRNet software. Results A total of 63 upregulated DEGs, 124 downregulated DEGs, four upregulated miRNAs and six downregulated miRNAs were identified. From the PPI network, the top 10 hub genes were identified, which were associated with platinum resistance. Our further qRT-PCR showed that seven hub genes (BUB1, KIF2C, NUP43, NDC80, NUF2, CCNB2 and CENPN) were differentially expressed in platinum-resistant ovarian cancer cells. Furthermore, the upstream transcription factors (TF) for upregulated DE-miRNAs were SMAD4, NFKB1, SMAD3, TP53 and HNF4A. Three overlapping downstream target genes (KIF2C, STAT3 and BUB1) were identified by miRNet, which was regulated by hsa-miR-494. Conclusions The TF-miRNA–mRNA regulatory pairs, that is TF (SMAD4, NFKB1 and SMAD3)-miR-494-target genes (KIF2C, STAT3 and BUB1), were established. In conclusion, the present study is of great significance to find the key genes of platinum resistance in ovarian cancer. Further study is needed to identify the mechanism of these genes in ovarian cancer.
研究领域:
肿瘤学
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分子生物学
·
基因组学
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DOI
PubMed
高影响因子
JOURNAL OF PHARMACEUTICAL AND BIOMEDICAL ANALYSIS
·
2020
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IF 3.209
Shuyu Mei,Bingru Liu,Xiaoli Xiong,Xu Hun
PtCuCo trimetallic alloys (PtCuCo-TAs) are synthesized by one-step reduction. The chemiluminescence (CL) properties of PtCuCo-TAs are studied systemically. PtCuCo-TAs show good catalyzing for luminol-H 2 O 2 system. A CL platform is developed for the detection of miRNA-21 using PtCuCo-TAs as nanozyme catalyzer. In the CL detection platform, H1 (Hairpin DNA1) is immobilized onto magnetic beads (MBs) firstly. In the presence of miRNA-21, H1 is opened. H2 (Hairpin DNA2) then hybridizes with H1. Meanwhile, a “cleat” in the end of miRNA-21 with a fewer bases complementary is formed to prevent miRNA-21 dissociating from H1. This miRNA-21 hybridizes to another H1. When cpDNA-PtCuCo-TAs which consisted with cDNA (Complementary strand of probe DNA) and pDNA-PtCuCo-TAs (PtCuCo-TAs labeled with probe DNA) are added, the ssDNA region of H1 reacts with the toehold domain of probe DNA and cDNA is released resulting pDNA-PtCuCo-TAs being captured. With this process repeatedly, a lot of pDNA-PtCuCo-TAs are captured onto MBs. After separation and washing, the precipitate and H 2 O 2 are put into the 96-well and luminol solution is injected. The CL signal is produced by PtCuCo-TAs catalyzing luminol-H 2 O 2 system. The amount of miRNA-21 is detected with CL signal. This CL platform performs with limit of detection 0.167 fM and has good selectivity over other RNA.
研究领域:
生物化学
·
分析化学
·
分子生物学
✓ 已验证
产品关联
DOI
PubMed
高影响因子
TALANTA
·
2023
·
IF 6.1
Dan Zhao,Jiutang Tang,Qin Tan,Xiaohong Xie,Xin Zhao,Dingpei Xing
Constructing an ultrasensitive CRISPR/Cas-based biosensing strategy is highly significant for the detection of trace targets. Here we presented a dual-amplified biosensing method based on CRISPR/Cas13a-triggered Cas12a, namely, Cas13a-12a amplification. As proof-of-principle, the developed strategy was used for miRNA-155 detection. The target bound to the Cas13a-crRNA complex and activated the cleavage activity of Cas13a for cleaving uracil ribonucleotides (rU) in the bulge structure of blocker strand (BS), resulting in the release of primer strand (PS) from the BS modified on magnetic beads. Then, the released PS activated the cleavage activity of Cas12a to cleave single-strand DNA reporter probes, producing a significantly increased fluorescent signal. The detection limit of the Cas13a-12a amplification using synthetic miRNA-155 was as low as 0.35 fM, which was much lower than that of the only Cas13a-based assay. The applied performance of this amplification strategy was verified by accurately quantifying miRNA-155 expression levels in different cancer patients. Therefore, the developed strategy offers a supersensitive and highly specific miRNAs sensing platform for clinical application.
研究领域:
分子生物学
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医学诊断
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生物技术
✓ 已验证
产品关联
DOI
PubMed
高影响因子
ANALYTICA CHIMICA ACTA
·
2025
·
IF 6
Fei Ding,Xiaomin Hang,Sen Tian,Wei Cao,Jingwen Wu,Li Wang
Primer exchange reaction (PER) is a novel and simple nucleic acid-templated extension technique that has recently attracted much attention in the field of biosensing. However, current PER reactions have shown relatively slow rates and low amplification performances, resulting in long assay times and limited detection sensitivities. Here we report a nicking endonuclease-mediated PER reaction (named NEPER) that rapidly releases amplified DNA products by adding a nicking endonuclease to hydrolyze the hybridized double-stranded DNA (dsDNA), and consequently has a maximum speed that is thirty orders of magnitude greater than the maximum for conventional PER. We further combined a CRISPR/Cas12a signal readout technique and developed a cascade NEPER-CRISPR/Cas12a method that can detect miRNA-155 with a limit of detection (LOD) down to 3.1 fM. We also show that the NEPER-CRISPR/Cas12a can be used to detect targets in serum samples.
研究领域:
分子生物学
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生物传感
·
诊断学
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产品关联
DOI
PubMed
高影响因子
APPLIED BIOCHEMISTRY AND BIOTECHNOLOGY
·
2025
·
IF 3.3
Zhang Xiuju,Nie Xiaowei,Yu Wenxiao,Du Guanchao,Liu Shaoming,Song Yewen
MicroRNA (miRNA) has emerged as a potential diagnostic marker for malignancies, such as prostate cancer; yet, few analytical approaches have been established for the simple and effective identification and quantification of miRNA. We depict here a fluorescent dumbbell probe utilizing DNA-silver nanoclusters (DNA-AgNCs) to facilitate the simple and label-free quantification of miRNA. The fluorescent probe was methodically engineered using a double-flapped dumbbell structure, wherein the 5′ flap was synthesized using DNA-AgNCs, and the 3′ flap was extended by a G-rich sequence. The DNA scaffold rigidifies the DNA-AgNCs and the G-rich sequence, bringing them into close contact, which leads to enhanced fluorescence due to the activation of DNA-AgNCs by the G-rich sequence. Upon the introduction of miRNA, the 5′ flap of the probe unfolds, liberating the G-rich sequence from the probe, disrupting the closeness between DNA-AgNCs and the G-rich region, and resulting in decreased fluorescence. The proposed method, by incorporating target recycling and polymerase/endonuclease-assisted cycles, enables sensitive detection of miRNA with a detection limit of 4.2 fM and demonstrates high specificity for monitoring target miRNA in clinical samples, offering a robust platform for miRNA monitoring and disease diagnosis. Graphical
研究领域:
生物化学
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分子生物学
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医学诊断
✓ 已验证
产品关联
DOI
PubMed
高影响因子
Biosensors-Basel
·
2023
·Vol. 13
·期 9
·854
·
IF 5.4
Yang Qing,Haobin Fang,Yuxing Yang,Yazhen Liao,Haiyu Li,Zhencui Wang,Jie Du
At present, a large number of studies have demonstrated that miRNAs can be used as biological indicators for the diagnosis and treatment of diseases such as tumours and cancer, so it is important to develop a new miRNA detection platform. In this work, miRNA-122 is used as the basis for targeting detection agents. We have designed an unlabelled DNA1 that undergoes partial hybridisation and has a 20 T base long strand. The fluorescent signal in this experiment is derived from copper nanoclusters (CuNCs) generated on the circular T-long strand of DNA1. At the same time, DNA1 is able to react with miRNA-122 and achieve hydrolysis of the part bound to miRNA-122 via the action of nucleic acid exonuclease III (Exo III), leaving a part of the DNA, called DNA3, while releasing miRNA-122 to participate in the next reaction, thus achieving circular amplification. DNA3 is able to react with DNA2, which is bound to streptavidin magnetic beads (SIBs) and separated from the reaction solution via the application of a magnetic field. Overall, this is a fluorescence signal reduction experiment, and the strength of the fluorescence signal from the copper nanoclusters can determine whether the target miRNA-122 is present or not. The degree of fluorescence reduction indicates how much DNA1, and thus the amount of target miRNA-122, has been hydrolysed. By evaluating the variations in the fluorescence signal under optimised conditions, we discovered that this method has good sensitivity, with a detection limit as low as 0.46 nM, better than many other previous works on fluorescence signal-based biosensors for miRNA detection. This technique offers high discrimination and selectivity and can serve as a persuasive reference for early diagnosis.Keywords:exonuclease III;miRNA-122;CuNCs;fluorescence reduction
研究领域:
分子生物学
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生物医学工程
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产品关联
DOI
PubMed
高影响因子
Chemosensors
·
2025
·Vol. 13
·期 4
·123
·
IF 3.7
Yanfei Zhang,Yiling Zhang,Runqi Ouyang,Zong Dai,Si-Yang Liu
MircroRNA (miRNA) exhibits abnormal expression in many cancer diseases, and the detection and analysis of miRNA are significant for the early diagnosis of diseases and research on miRNA functions. In this work, we construct a UV-triggered DNAzyme (UTD) nanosensor for the early detection of miRNA in tumor cells. As the nanodevice was delivered into cells and irradiated by UV light, the controllable imaging of miRNA in living cells was achieved. This method effectively avoids false signal issues, providing a new strategy for high-spatiotemporal-resolution imaging of miRNA in living cells.
研究领域:
分子生物学
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医学诊断
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生物技术
✓ 已验证
产品关联
DOI
高影响因子
Journal of Analytical Science and Technology
·
2024
·Vol. 15
·期 1
·1-8
·
IF 2.5
Li Yunxiao,Wang Qiuxia,Wang Yali
The abnormally expressed microRNAs (miRNAs) serve as crucial indicators for disease diagnosis and are strongly associated with the progression of various diseases. The quantification of miRNAs is highly significant for the clinical diagnosis and treatment of various types of cancers. This study utilizes the CRISPR/Cas13a system, which combines CRISPR RNA (crRNA) and CRISPR-associated Cas13a, to recognize the miRNA directly and specifically, thus activating the trans-cleavage activity of Cas13a. By integrating the CRISPR-Cas13a system with an exonuclease-iii (Exo-iii)-assisted chain cleavage of silver ions (Ag+)-aptamer and an Ag+ -based color reaction, a method for the detection of miRNA that is specific and sensitive is developed. This approach demonstrated exceptional sensitivity in detecting miRNA, with a low detection limit of 5.12 fM. Moreover, the suggested method was effectively utilized to precisely measure the amount of miRNA in intricate biological samples, demonstrating its practical applicability. Furthermore, due to its elevated sensitivity and simple probe design and fluorophore labeling, the suggested colorimetric technique holds great potential for use in clinical diagnostics.
研究领域:
分子生物学
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医学诊断
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生物技术
✓ 已验证
产品关联
DOI
高影响因子
ACS Applied Materials & Interfaces
·
2023
·
IF 9.5
Shaoqiong Feng,Hanjun Chen,Ziao Hu,Tingting Wu,Zhihong Liu
MicroRNA (miRNA) is a promising biomarker for the diagnosis, monitoring, and prognostic evaluation of diseases, especially cancer. The existing miRNA detection methods usually need external instruments for quantitative signal output, limiting their practical applications in point-of-care (POC) settings. Here, we propose a distance-based biosensor through a responsive hydrogel, in combination with a CRISPR/Cas12a system and target-triggered strand displacement amplification (SDA) reaction for visual quantitative and sensitive measurement of miRNA. The target miRNA is first converted into plenty of double-stranded DNA (dsDNA) via target-triggered SDA reaction. Then, the dsDNA products trigger the collateral cleavage activity of CRISPR/Cas12a, leading to the release of trypsin from magnetic beads (MBs). The released trypsin can hydrolyze gelatin, and hence the permeability of gelatin-treated filter paper is increased, resulting in a visible distance signal on a cotton thread. Using this system, the concentration of the target miRNA can be quantified visually without any assistance of instruments, and a detection limit of 6.28 pM is obtained. In addition, the target miRNA in human serum samples and cell lysates can also be detected accurately. Owing to the characteristics of simplicity, sensitivity, specificity, and portability, the proposed biosensor provides a new tool for miRNA detection and holds great promise in POC applications.
研究领域:
生物技术
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分子诊断
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医学检验
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DOI
PubMed
高影响因子
FEBS LETTERS
·
2015
·
IF 3.169
Chen Xie,Wei Chen,Mengdie Zhang,Qiuxian Cai,Weiyi Xu,Xiaodi Li,Songshan Jiang
Despite extensive investigation into the role of let-7 miRNAs in pathological tumor processes, their involvement in the DNA damage response remains unclear. Here we show that most let-7 family members down-regulate MDM4 expression via binding to MDM4 mRNA at a conserved DNA sequence. Expression of exogenous let-7 miRNA mimics decreased MDM4 protein but not mRNA levels. Several DNA damage reagents increased let-7 expression, thereby decreasing MDM4 protein levels in glioma cells. Inhibition of endogenous let-7 with antisense RNAs rescued MDM4 protein levels with or without MG132, a proteasome-dependent degradation inhibitor. An MDM4 mutation identified in a glioma patient was associated with loss of the putative MDM4 target site. Therefore, let-7 binding to MDM4 is implicated in the DNA damage response.
研究领域:
癌症生物学
·
分子生物学
·
遗传学
✓ 已验证
DOI
PubMed
高影响因子
TALANTA
·
2025
·
IF 6.1
Xiaofang Liu,Qun Wang,Huan Liu,Yong Zhang,Mei Yang,Danqun Huo,Changjun Hou
Timely detection of breast cancer not only positively influences patient treatment and life quality, but also possesses far-reaching implications for public health and medical research. And miRNAs, as emerging breast cancer-related biomarkers, demonstrate heightened sensitivity to disease progression. In this study, the same precursor was utilized to prepare micromaterials with large and small cavities, this was the safeguard for sensing probes and signals. And the signal amplification process was accomplished with strand displacement assistance by target excitation. Encouragingly, it was successfully and accurately detected miRNA-21 and miRNA-155 within 10 fmol/L - 10 nmol/L. Due to the unique sensing design, the signal can be read out not only by small fluorescent devices, but also by portable glucometers. The biosensor features several excellent working performances and illuminates resource-poor areas by equipping the most common glucometers. So, it is a sensitive, portable and promising biosensor.
研究领域:
分子生物学
·
医学检测
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生物传感技术
✓ 已验证
产品关联
DOI
PubMed
高影响因子
JOURNAL OF WUHAN UNIVERSITY OF TECHNOLOGY-MATERIALS SCIENCE EDITION
·
2024
·Vol. 39
·期 4
·1048-1052
·
IF 1.3
Yu Dan,Ye Liyuan,Li Binbin,Mou Fangzhi,Yin Yixia
pH-responsive charge reversal loaded miRNA nanocomposite was prepared by electrostatic self-assembly. The morphology, particle size and zeta potential of the nanocomposites were analyzed by transmission electron microscopy and dynamic light scattering. The synthesis of the polymer was analyzed by 1 H-NMR. The zeta-potential changes and cellular uptake effects of the nanocomplexes under different pH environments were investigated. The experimental results show that the surface morphology of the nanocomposite is spherical, and the average particle size is about 135 nm. As the pH value of the solution gradually decreases, the surface charge of the nanocomposite reverses from negative charge to positive charge (from −9.4 to +17.1 mV). Cellular uptake mediated by pH-responsive nanocomposite is selective for tumor cells, and the cellular uptake effect in tumor cells at pH 6.5 was approximately 3 times higher than that at pH 7.4. This pH responsive charge reversal nanocomposite has promising application prospects for gene delivery in the weak acid environment of tumors.
研究领域:
生物材料学
·
分子生物学
·
药物递送系统
✓ 已验证
产品关联
DOI
高影响因子
危险品化学品经营许可证（带存储）