Molecular Medicine Reports
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2021
·Vol. 25
·期 2
·1-13
·
IF 2.952
Hongbin Peng,Pingjiang Ge
It has been reported that long non‑coding RNA HLA complex group 18 (HCG18) is involved in the progression of cancer, acting as an oncogenic gene. The aim of the present study was to investigate the mechanism underlying the action of HCG18 in laryngeal and hypopharyngeal squamous cell carcinoma (LHSCC). The expression levels of HCG18, microRNA (miR)‑133b and fibroblast growth factor receptor 1 (FGFR1) in LHSCC tissues and transfected LHSCC cells were evaluated by reverse transcription‑quantitative PCR or immunohistochemistry. The viability, migration and invasion of transfected LHSCC cells were detected by Cell Counting Kit‑8, wound healing and Transwell assays, respectively. The targeting relationships of HCG18, miR‑133b and FGFR1 were predicted by bioinformatics analysis and confirmed using a dual‑luciferase reporter assay. Moreover, the expression levels of FGFR1, phosphorylated (p)‑PI3K, PI3K, p‑AKT, AKT, p53, Bax and Bcl‑2 in transfected LHSCC cells were measured by western blotting. It was found that the expression levels of HCG18 and FGFR1 were upregulated, but those of miR‑133b were downregulated in LHSCC tissues. Short hairpin RNA (sh) HCG18 and miR‑133b mimic inhibited LHSCC cell viability, while enhancing miR‑133b expression. HCG18 could competitively bind with miR‑133b. Moreover, the miR‑133b inhibitor promoted cell viability, migration, invasion and the expression levels of Bcl‑2, p‑PI3K and p‑AKT, but inhibited the expression levels of p53 and Bax, which were abrogated by shHCG18. miR‑133b could competitively bind with FGFR1, and the miR‑133b mimic decreased the expression level of FGFR1 in transfected LHSCC cells. shFGFR1 promoted the expression levels of p53 and Bax, while inhibiting viability, migration, invasion and Bcl‑2, p‑PI3K and p‑AKT expression in LHSCC cells. In conclusion, the current results indicated that HCG18 facilitated the progression of LHSCC by upregulating FGFR1 via miR‑133b. The present study evaluated the mechanism with regards to the action of HCG18 in LHSCC, and these experimental results may provide novel evidence for targeted therapy of LHSCC.
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TALANTA
·
2023
·
IF 6.1
Gefei Huang,Feng Zhang,Dongying Xie,Yiming Ma,Pengxi Wang,Guodong Cao,Leijian Chen,Siyi Lin,Zhongying Zhao,Zongwei Cai
Although next-generation sequencing technology has been used to delineate RNA modifications in recent years, the paucity of appropriate converting reactions or specific antibodies impedes the accurate characterization and quantification of numerous RNA modifications, especially when these modifications demonstrate wide variations across developmental stages and cell types. In this study, we developed a high-throughput analytical platform coupling ultra-performance liquid chromatograph (UPLC) with complementary mass spectrometry (MS) to identify and quantify RNA modifications in both synthetic and biological samples. Sixty-four types of RNA modifications, including positional isomers and hypermodified ribonucleosides , were successfully monitored within a 16-min single run of UPLC–MS. Two independent methods to cross-validate the purity of RNA extracted from Caenorhabditis elegans ( C. elegans ) were developed using the coexisting C. elegans and Escherichia coli ( E. coli ) as a surveillance system. To test the validity of the method, we investigated the RNA modification landscape of three model organisms, C. elegans , E. coli , and Arabidopsis thaliana ( A. thaliana ). Both the identity and molarity of modified ribonucleosides markedly varied among the species. Moreover, our platform is not only useful for exploring the dynamics of RNA modifications in response to environmental cues (e.g., cold shock) but can also help with the identification of RNA-modifying enzymes in genetic studies. Cumulatively, our method presents a novel platform for the comprehensive analysis of RNA modifications, which will be of benefit to both analytical chemists involved in biomarker discovery and biologists conducting functional studies of RNA modifications.
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BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS
·
2018
·
IF 2.559
Li-Xin Tang,Guo-Hua Chen,Huan Li,Ping He,Yan Zhang,Xue-Wen Xu
Long noncoding RNA (OGFRP1) has been reported to be involved in the progression of non-small cell lung cancer (NSCLC). However, the expression pattern, functions and molecular mechanisms of OGFRP1 in NSCLC remains unclear. In the present study, we found that OGFRP1 expression was significantly up-regulated in both NSCLC tissues and cell lines, and the upregulation of OGFRP1 expression is a powerful predictor of advanced clinical stage, lymph nodes metastasis and poor prognosis for NSCLC patients. Loss-of-function assay indicated that knockdown of OGFRP1 inhibited proliferation, migration and invasion, and induced apoptosis in vitro. Mechanistically, OGFRP1 could directly bind to miR-124-3p and effectively act as a competing endogenous RNA (ceRNA) for miR-124-3p to promote the expression of the target gene LYPD3. Taken together, OGFRP1 contributed to progression of NSCLC at least partly through upregulating LYPD3 expression by sponging miR-124-3p, indicating that OGFRP1 may be a novel prognostic biomarker and therapeutic target in NSCLC.
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INTERNATIONAL IMMUNOPHARMACOLOGY
·
2024
·
IF 4.8
Zhongkai Zhao,Zeyu Miao,Yuyang Hou,Yifan Zhong,Xiaorong Zhang,Xuedong Fang
Background A newly identified type of cell death due to intracellular copper accumulation is known as cuproptosis and RNA methylation is a post-transcriptional modification mechanism, both of which perform vital roles in the immune microenvironment of colorectal cancer (CRC), but the link between the two needs more research. Methods TCGA database provided RNA-seq data and details clinically of CRC samples. Cuproptosis-related RNA methylation regulators (CRRMRs) were identified by correlation analysis. We screened 6 CRRMRs for prognostic model construction by employing LASSO-Cox regression analysis and calculated risk scores by CRRMRs (CuMS). GSE39582 and GSE38832 cohort were used as external validation sets. This research concentrated on the connection between the prognostic model and somatic mutation, anti-cancer drug sensitivity, immune infiltration, immune checkpoint expression. In addition, we investigated the differential expression of YTHDC2 in epithelial cell subpopulations by single-cell analysis with GSE166555, calculated cuproptosis scores and performed pathway enrichment. In vitro experiments were performed to explore the consequences of knockdown of YTHDC2 on CRC cell proliferation and migration, as well as changes in CRC cell viability in response to elesclomol after knockdown of YTHDC2. In vivo experiments, we constructed the cell line-derived xenograft model to further validate the results of the in vitro experiments. Results The prognosis of CRC can be predicted by CuMS, which GSE39582 and GSE38832 confirmed. Two CuMS groups showed different tumor mutation burden (TMB) and immune infiltration. CuMS was connected to emerging immune checkpoints CD47 and PVR, therefore, it can be clinically complementary to TMB and microsatellite instability (MSI) status. In single-cell analysis, a subpopulation of epithelial cells with high YTHDC2 expression had a high cuproptosis score. In vitro experiments, knocking down YTHDC2 promoted cell proliferation and migration in CRC, and weaken the inhibitory effect of elesclomol and elesclomol-Cu on cell viability, which in vivo experiments validated. Conclusion We developed a prognostic model constructed by 6 CRRMRs to assess overall survival and immune microenvironment of CRC patients. YTHDC2 might regulate cuproptosis in multiple ways.
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MICROCHEMICAL JOURNAL
·
2024
·
IF 4.9
Yuanxun Gong,Jiayi Zhang,Huiying Liang,Yanling Wang,Zichun Song,Jihua Wei,Qianli Tang,Kai Zhang,Xianjiu Liao
Matrix metalloproteinases (MMPs) are critical for the degradation of extracellular matrix components, which is essential for processes such as tissue remodeling, wound healing, and angiogenesis. MMP-2, in particular, degrades type IV collagen and is associated with diseases such as cancer and cardiovascular conditions, making it a significant biomarker. This study presents a novel electrochemical biosensor for the detection of MMP-2. The biosensor combines the specificity of a programmable RNA detection system with the amplification capabilities of T7 RNA polymerase and CRISPR/Cas12a. In the absence of MMP-2, Peptide Nucleic Acid (PNA) adsorbs on the DNA template, preventing RNA amplification and resulting in a low electrochemical signal. When MMP-2 is present, it cleaves a specific peptide, releasing the DNA template and enabling RNA synthesis. This activation leads to the cleavage of single-stranded DNA probes, increasing the electrochemical signal. The biosensor demonstrates high sensitivity with a detection limit of 0.496 fM for MMP-2 and excellent specificity, effectively distinguishing MMP-2 from other proteins. It successfully detected MMP-2 in biological samples, such as liver cancer cell supernatants, indicating its potential for clinical diagnostics. This innovative biosensor offers a powerful tool for the early detection and monitoring of diseases associated with MMP-2 activity.
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ANALYTICAL CHEMISTRY
·
2025
·
IF 6.7
Yuting Li,Yunqing Tang,Lu Qian,Yanqin Zhang,Yuewen Zhai,Jinnan Zhang,Yu Li,Yanfang Deng,Juequan Li,Yuhang Li,Bing Xue,Yunlong Liu,Siwen Li
RNA in small extracellular vesicles (sEVs) from plasma is a reliable biomarker for disease diagnosis. However, the cumbersome steps and operator dependency for extraction and analysis of sEVs RNA in plasma have limited their application. Here, we developed a multisatellites@planets molecular imprinting detection platform for sEVs capture and RNA analyzation in a one-step process directly from a small amount of plasma. In this platform, magnetic beads as “planet” templates were utilized for size recognition of sEVs, and Au nanoparticles as “multi-satellites” templates were used to immobilize liposome probes for sEVs grasp and RNA detection through membrane fusion. As for the detection accuracy, the results from our platform demonstrate a high level of concordance with qRT-PCR (R2 value of 0.9937). Moreover, we successfully used 10 μL of plasma to analyze sEVs RNA, effectively differentiating healthy donors and lung cancer patients with an accuracy of 91.18% (combined MALAT1 and miR-21). This novel platform provides a pretreatment-free, user-friendly, highly accurate, and sensitive strategy for detecting the RNA of sEVs in plasma, indicating its potential capability for disease diagnosis.
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Research
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2025
·
IF 10.7
Han Wang,Zhouyi Gan,Yueyue Wang,Dingmeng Hu,Lexiang Zhang,Fangfu Ye,Ping Duan
Endometriosis is marked by the ectopic growth, spread, and invasion of endometrial tissue beyond the uterus, resulting in recurrent bleeding, pain, reproductive challenges, and the formation of nodules or masses. Despite advancements in detection methods like ultrasound and laparoscopy, these techniques remain limited by low specificity and invasiveness, underscoring the need for a highly specific, noninvasive in vitro diagnostic method. This study investigates the potential of using menstrual blood as a noninvasive diagnostic sample for endometriosis by targeting genetic and inflammatory markers associated with endometriosis lesions. A novel digital droplet enzyme-linked immunosorbent assay (ddELISA) was developed, leveraging SiO 2 nanoparticles for the femtomolar-sensitive detection of inflammatory cytokines (OPN, IL-10, IL-6) in menstrual blood. Single-cell RNA sequencing revealed differentiation patterns across endometrial tissues and menstrual blood, affirming that menstrual blood replicates key inflammatory and immune properties of endometriosis. Furthermore, endometriosis menstrual blood endometrial cells derived from human menstrual blood displayed similar properties to endometrial stromal cells in endometriosis lesions, validating menstrual blood as a suitable in vitro diagnostic sample. In contrast to traditional ELISA, ddELISA supports multi-target detection with enhanced sensitivity and reduced processing time, allowing precise biomarker analysis from minimal sample volumes. Our ddELISA-based approach shows promise as a rapid, accessible, and accurate diagnostic tool for endometriosis, with potential for practical clinical application.
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Frontiers in Physiology
·
2017
·
IF 4.134
Jiang Jianru,Xu Yili,Lin Xinda
Ovarian development plays an important role in the life history of insects and is crucial for control of the insect population. The metamorphosis of an insect is precisely regulated by the interaction of the juvenile hormone and ecdysone. To understand the role of NlBr and NlKr-h1 in ovary development, we used RNA interference (RNAi) to down-regulate the expression of Broad-Complex (Br) and Krüppel homolog 1 (Kr-h1), two important down-stream transcription factors of juvenile hormone and ecdysone signaling. We further investigated their effects on metamorphosis and ovary development. The results showed that both NlBr and NlKr-h1 are induced by ecdysone. The down-regulation of NlBr and NlKr-h1 alone or together by RNAi is more effective than the topical application of ecdysone on the number of ovarioles, suggesting the necessity of NlBr and NlKr-h1 in determining the number of ovarioles. The ovarian grade was significantly increased/decreased by the topical application of ecdysone and down-regulation of NlBr and NlKr-h1. The pre-oviposition period was also increased. When NlBr and NlKr-h1 were down-regulated together, the ovary grade was not significantly different compared to the control (dsGFP), indicating that the development of the ovary is under the control of both NlBr and NlKr-h1. The interaction between the NlBr and NlKr-h1 on the number of ovarioles and the development of the ovary indicates cross-talk between both juvenile hormone and ecdysone signaling at the transcription level in the brown planthopper. Both genes are nuclear transcription factors and may regulate signaling via down-stream genes. These results would help to both enhance the current understanding of the regulatory mechanism of the interaction between juvenile hormone and ecdysone signaling pathways during ovarian development and to design chemicals to control pests.
研究领域:
昆虫学
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分子生物学
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ACS Nano
·
2025
·
IF 16
Kai Yu,Lidan Fu,Yu Chao,Xiaodong Zeng,Yonggang Zhang,Yuanyuan Chen,Jialu Gao,Binchun Lu,Hua Zhu,Lijuan Gu,Xiaoxing Xiong,Zhenhua Hu,Xuechuan Hong,Yuling Xiao
Small interfering RNA (siRNA) targeting the NOD-like receptor family pyrin domain-containing 3 (NLRP3) inflammasome has emerged as a promising therapeutic strategy to mitigate infarct volume and brain injury following ischemic stroke. However, the clinical translation of siRNA-based therapies is significantly hampered by the formidable blood-brain barrier (BBB), which restricts drug penetration into the central nervous system. To address this challenge, we have developed an innovative long-circulating near-infrared II (NIR-II) nanoparticle platform YWFC NPs, which is meticulously engineered to enhance BBB transcytosis and enable efficient delivery of siRNA targeting NLRP3 (siNLRP3@YWFC NPs) in preclinical models of ischemic stroke. Furthermore, we integrated advanced deep learning neural network algorithms to optimize in vivo NIR-II imaging of the cerebral infarct penumbra, achieving an improved signal-to-background ratio at 72 h poststroke. In vivo studies employing middle cerebral artery occlusion (MCAO) mouse models demonstrated that image-guided therapy with siNLRP3@YWFC NPs, guided by prolonged NIR-II imaging, resulted in significant therapeutic benefits.
研究领域:
神经科学
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药理学
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医学影像学
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MICROCHIMICA ACTA
·
2020
·Vol. 187
·期 4
·1-9
·
IF 6.232
Cao Xiaodong,Zhang Kairui,Yan Wuwen,Xia Zihao,He Shudong,Xu Xuan,Ye Yongkang,Wei Zhaojun,Liu Songqin
A selective and sensitive fluorescence biosensor is described for determination of microRNA-167 using fluorescent resonant energy transfer (FRET) strategy. The FRET system comprises carbon dots (CDs, donor) labeled with probe DNA (pDNA) and polydopamine (PDA)-coated Fe 3 O 4 nanoparticles (Fe 3 O 4 @PDA NPs, acceptor). The CDs-pDNA can be absorbed onto the surface of Fe 3 O 4 @PDA NPs because of the strong π interaction between pDNA and PDA. With the enhanced adsorption ability of Fe 3 O 4 @PDA NPs by Ca 2+ , the fluorescence intensity of CDs at 445 nm (excitation at 360 nm) is quenched. In presence of microRNA-167, the hybridized complex of CDs-pDNA-microRNA-167 will be released from the surface of Fe 3 O 4 @PDA NPs due to the weak π interaction of the complex and PDA. This results in the fluorescence recovery of CDs. By application of twice-magnetic separation, the biosensor shows a wide linear range of 0.5–100 nM to microRNA-167 with a 76 pM detection limit. The method was applied to the determination of microRNA-167 in samples of total microRNA extractions from A. thaliana seedlings, and the recoveries ranged from 96.4 to 98.3%.
研究领域:
生物化学
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分子生物学
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SENSORS AND ACTUATORS B-CHEMICAL
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2021
·
IF 7.46
Xingjie Wu,Hong Yang,Wei Li,Ling Tao,Qingteng Wang,Yingzu Jin,Xuecheng Sun,Xiangchun Shen
The prompt diagnosis of acute myocardial infarction (AMI) for patients with acute ischemic-type chest pain is critical for timely AMI treatment and lowering its casualty rate. In this article, we developed a multiparametric detection technique for AMI related microRNAs (miRNAs), which is based on the polymerization induced fluorescence-off (PIFO) signal amplification technique. PIFO can be implemented on a specially designed microfluidic chip equipping a fluorescence sensor, which is able to profile 3 miRNAs simultaneously in 45 min with a detection limit of 6100 miRNAs copies. Comparing with other polymerization-based signal amplification techniques, PIFO is rapid, oxygen-independent, multiparametric, and can be performed in ambient environment. The miniaturized PIFO device can be further applied to analyze AMI related miRNAs in clinical serum samples, and displays excellent performance in distinguishing AMI patients from patients with other cardiovascular diseases. Moreover, PIFO shows a high accuracy for AMI diagnosis among patients with low hypersensitivity troponin I level (< 0.1 ng mL −1 ) as a result of the early and fast respond to AMI onset by miRNAs.
研究领域:
医学诊断
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分子生物学
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生物传感器
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Journal of Translational Medicine
·
2024
·Vol. 22
·期 1
·1-19
·
IF 6.1
Xu Haotian,Zheng Yue,Wu Jiaxi,Zhang Ruirui,Zhao Qingyun,Chen Sixian,Peng Wenyi,Cai Dunyu,Gao Yihong,Chen Xingcai,Li Deqing,yuan Shengyi,Li Gang,Nan Aruo
Lung cancer is the primary cause of cancer-related death worldwide, and its global incidence and mortality rates remain high. The differential expression of circular RNAs (circRNAs) can affect the development of cancer, but the mechanisms by which circRNAs regulate lung cancer progression remain unclear. In this study, we identified circSORBS1, a circRNA that has not been previously described in lung cancer and is significantly underexpressed in lung cancer tissues, blood and cell lines, and the low expression of circSORBS1 correlated with tumour grade and prognosis. In vitro and in vivo functional experiments revealed that circSORBS1 overexpression inhibited cell proliferation and migration while enhancing apoptosis. Mechanistically, circSORBS1 acts as a sponge for miR-6779-5p, indirectly inhibiting RUFY3 mRNA degradation. Simultaneously, it binds to RUFY3 mRNA to enhance its stability. This dual regulatory mechanism leads to an increase in RUFY3 protein levels, which ultimately activates the YWHAE/BAD/BCL2 apoptotic signalling pathway and suppresses lung cancer progression. Our findings not only increase the knowledge about the regulatory pattern of circRNA expression but also provide new insights into the mechanisms by which circRNAs regulate lung cancer development.
研究领域:
肿瘤学
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分子生物学
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癌症生物学
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基因组学
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