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MnO2 Nanosheet-mediated Ratiometric Fluorescence Biosensor for MicroRNA Detection and Imaging in Living Cells
摘要： MicroRNA (miRNA) plays significant roles in cell proliferation, differentiation and apoptosis, and has been considered to be valuable biomarker for cancer. Accurate and sensitive detection of miRNA is crucially significant for cancer diagnosis and treatment. Here, a MnO2 nanosheet-mediated ratiometric fluorescence biosensor was designed for miRNA detection and imaging in living cells. It contained MnO2 nanosheets acting as DNA carrier, and fluorescent donor (FAM)-labeled hairpin H1 (recognition probe) and fluorescent acceptor (TAMRA)-labeled hairpin H2 (amplification probe). When the biosensor entered cell by endocytosis, MnO2 nanosheets were degraded to Mn2+ via intracellular glutathione (GSH) and the adsorbed hairpins H1 and H2 were released. The intracellular target miRNA-21 hybridized with the recognition unit of H1 to initiate catalyzed hairpin assembly (CHA) and a large amount of H1-H2 duplexes were produced. This brought fluorescent donor FAM and fluorescent acceptor TAMRA into close proximity to produce fluorescence resonance energy transfer (FRET), inducing a ratiometric fluorescent response (donor signal decreased and acceptor signal enhanced) for miRNA-21 detection. Furthermore, this method could be applied to differentiate the expression levels of miRNA-21 in HeLa, HepG-2 and L02 cells. These results indicated that the proposed method possessed great potential in the early diagnosis of miRNA-related diseases.
关键词： MicroRNA detection,MnO2 nanosheets,Ratiometric,Cell imaging
更新于2025-11-21 11:24:58
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Cationic porphyrins with large side arm substituents as resonance light scattering ratiometric probes for specific recognition of nucleic acid G-quadruplexes
摘要： Specific G-quadruplex-probing is crucial for both biological sciences and biosensing applications. Most reported probes are focused on fluorescent or colorimetric recognition of G-quadruplexes. Herein, for the first time, we reported a new specific G-quadruplex-probing technique—resonance light scattering (RLS)-based ratiometric recognition. To achieve the RLS probing of G-quadruplexes in the important physiological pH range of 7.4-6.0, four water soluble cationic porphyrin derivatives, including an unreported octa-cationic porphyrin, with large side arm substituents were synthesized and developed as RLS probes. These RLS probes were demonstrated to work well for ratiometric recognition of G-quadruplexes with high specificity against single- and double-stranded DNAs, including long double-stranded ones. The working mechanism was speculated to be based on the RLS signal changes caused by porphyrin protonation that was promoted by the end-stacking of porphyrins on G-quadruplexes. This work adds an important member in G-quadruplex probe family, thus providing a useful tool for studies on G-quadruplex-related events concerning G-quadruplex formation, destruction and changes in size, shape and aggregation. As a proof-of-concept example of applications, the RLS probes were demonstrated to work well for label-free and sequence-specific sensing of microRNA. This work also provides a simple and useful way for the preparation of cationic porphyrins with high charges.
关键词： G-quadruplex,ratiometric recognition,microRNA sensing,resonance light scattering,cationic porphyrin
更新于2025-11-19 16:56:35
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Photoelectrochemical biosensor for microRNA detection based on a MoS2/g-C3N4/black TiO2 heterojunction with Histostar@AuNPs for signal amplification
摘要： Herein, a novel photoelectrochemical (PEC) biosensor was developed for the ultrasensitive detection of microRNA-396a based on a MoS2/g-C3N4/black TiO2 heterojunction as the photoactive material and gold nanoparticles carrying Histostar antibodies (Histostar@AuNPs) for signal amplification. Briefly, MoS2/g-C3N4/black TiO2 was deposited on an indium tin oxide (ITO) electrode surface, after which gold nanoparticles (AuNPs) and probe DNA were assembled on the modified electrode. Hybridization with miRNA-396a resulted in a rigid DNA:RNA hybrid being formed, which was recognized by the S9.6 antibody. The captured antibody can further conjugate with the secondary IgG antibodies of Histostar@AuNPs, thereby leading to the immobilization of horse radish peroxidase (HRP). In the presence of HRP, the oxidation of 4-chloro-1-naphthol (4-CN) by H2O2 was accelerated, producing the insoluble product benzo-4-chlorohexadienone on the electrode surface and causing a significant decrease in the photocurrent. The developed biosensor could detect miRNA-396a at concentrations from 0.5 fM to 5000 fM, with a detection limit of 0.13 fM. Further, the proposed method can also be used to investigate the effect of heavy metal ions on the expression level of miRNAs. Results suggest that the biosensor developed herein offers a promising platform for the ultrasensitive detection of miRNA.
关键词： S9.6 antibody,Histostar@AuNPs,MicroRNA detection,MoS2/g-C3N4/black TiO2 heterojunction,Photoelectrochemical biosensor
更新于2025-11-14 17:04:02
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A novel photoelectrochemical biosensor for the sensitive detection of dual microRNAs using molybdenum carbide nanotubes as nanocarriers and energy transfer between CQDs and AuNPs
摘要： Herein, a novel photoelectrochemical (PEC) biosensor was developed for the ultrasensitive detection of dual microRNAs (miRNAs), with the detection being based on energy transfer (ET) between carbon quantum dots (CQDs) and gold nanoparticles (AuNPs). The PEC platform consisted of a CQDs@Mo2C nanotube modified ITO electrode. Two hairpin probes (H1 and H2) carrying the Au NPs were used “switch off” and “switch on” the PEC signal of the CQDs, with a close approach of the tagged AuNPs to the CQDs quenching the PEC signal. The introduction of different miRNAs (miRNA-159b and miRNA-166a) altered the interparticle distance between the AuNPs and CQDs, thereby affecting the intensity of the PEC response. This approach allowed the highly sensitive detection of both miRNA-159b and miRNA-166a. The linear range of the biosensor for miRNA-159b and miRNA-166a detection were 0.5–5000 fM, with low detection limits of 0.15 fM and 0.21 fM, respectively. To our knowledge, this is the first reported CQDs-based ET biosensor for the PEC detection of dual miRNAs. Results suggest that this approach offers a promising platform for the ultrasensitive detection of multiple miRNAs.
关键词： MicroRNA detection,AuNPs,Photoelectrochemistry,Energy transfer,CQDs@Mo2C
更新于2025-11-14 17:03:37
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Morpholino Oligonucleotide Cross-Linked Hydrogels as Portable Optical Oligonucleotide Biosensors
摘要： Morpholino Oligonucleotides (MOs), an uncharged DNA analogue, are functionalized with an acrylamide moiety and incorporated into polymer hydrogels as responsive crosslinks for microRNA sequence detection. The MO crosslinks can be selectively cleaved by a short target analyte single-stranded DNA (ssDNA) sequence based on microRNA, inducing a distinct swelling response measured optically. The MO crosslinks offer significant improvement over DNA based systems through improved thermal stability, no salt requirement and 1000-fold improved sensitivity over a comparative biosensor, facilitating a wider range of sensing conditions. Analysis was also achieved using a mobile phone camera, demonstrating portability.
关键词： microRNA,Biosensor,Oligonucleotide sensor,Morpholino oligonucleotide,Hydrogel
更新于2025-09-23 15:23:52
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Baicalin protects human retinal pigment epithelial cell lines against high glucose-induced cell injury by up-regulation of microRNA-145
摘要： Background: Diabetic retinopathy (DR) is a common complication of diabetes mellitus, which is a major reason of blindness. Baicalin (BAI) is a flavonoid extracted from Scutellaria baicalensis, whose pharmacological characterizes have been widely reported in various diseases. However, it remains unclear the effect of BAI on DR. The study aimed to confirm the protective effect of BAI on DR. Methods: ARPE-19 cells and HRMECs were exposed to the high glucose (HG) environment to construct a cell injury model. After treatment with HG and BAI, cell viability, apoptosis, inflammatory cytokines and ROS generations were determined in ARPE-19 cells and HRMECs. Subsequently, miR-145 inhibitor and its negative control were transfected into ARPE-19 cells, and the regulatory effects on HG-and BAI-co-treated cells were detected. NF-κB and p38MAPK signaling pathways were finally examined to state the underling mechanism. Results: HG treatment significantly induced ARPE-19 cells and HRMECs injury in vitro. BAI significantly promoted cell proliferation, reduced apoptosis, as well as inhibited the release of IL-1β, IL-6, IL-8 and ROS in HG- injured ARPE-19 cells and HRMECs. Additionally, the expression level of miR-145 was up-regulated in HG-and BAI-co-treated cells. More importantly, miR-145 inhibition reversed the protective effect of BAI on HG- injured ARPE-19 cells. Besides, we observed that BAI inhibited the activation of NF-κB and p38MAPK pathways by up-regulating miR-145. Conclusions: Results demonstrated that BAI exhibited the protective effect against HG-induced cell injury by up-regulation of miR-145.
关键词： microRNA-145,diabetic retinopathy,p38MAPK,NF-κB,baicalin
更新于2025-09-23 15:23:52
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Identification and imaging of miR-155 in the early screening of lung cancer by targeted delivery of octreotide-conjugated chitosan-molecular beacon nanoparticles
摘要： Lung cancer is still the most common cancer globally. Early screening remains the key to improve the prognosis of patients. There is currently a lack of specific and sensitive methods for early screening of lung cancer. In recent years, studies have found that microRNA plays an important role in the occurrence and development of lung cancer and become a biological target in the early diagnosis of lung cancer. In this study, lung cancer cells, subcutaneous xenografts of lung cancer in nude mice, and Lox-Stop-lox K-ras G12D transgenic mice were used as models. The transgenic mice displayed the dynamic processes from normal lung tissue to atypical hyperplasia, adenomas, carcinoma in situ and lung adenocarcinoma. It was found that miR-155 and somatostatin receptor 2 (SSTR2) were expressed in all the disease stages of transgenic mice. Through molecular beacon (MB) technology and nanotechnology, chitosan-molecular beacon (CS-MB) nanoparticles and targeted octreotide (OCT) were conjugated and synthesized. The octreotide-conjugated chitosan-molecular beacon nanoparticles (CS-MB-OCT) can specifically bind to SSTR2 expressed by the lung cancer cells to achieve the goal of identification of lung cancer cells and imaging miR-155 in vivo and in vitro. Fluorescence imaging at different disease stages of lung cancer in Lox-Stop-lox K-ras G12D transgenic mice was performed, and could dynamically monitor the occurrence and development of lung cancer by different fluorescence intensity ranges. The current research, in turn, provides new idea, new method, and new technology for the early screening of lung cancer.
关键词： chitosan nanoparticles,molecular imaging,molecular beacon,Lung cancer,microRNA-155
更新于2025-09-23 15:23:52
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MicroRNA-382 inhibits cell proliferation and invasion of retinoblastoma by targeting BDNF-mediated PI3K/AKT signalling pathway
摘要： It has previously been demonstrated that multiple microRNAs (miRNAs or miRs) are aberrantly expressed in retinoblastoma (RB) and contribute to RB initiation and progression. miR-382 has been revealed to be aberrantly expressed and therefore exhibits a key role in the progression of various types of cancer. However, the expression pattern, functional roles and underlying molecular mechanism of miR-382 in RB remain unknown. The present study investigated the expression levels of miR-382 and its effects on RB cells and the underlying regulatory mechanism of its action. It was demonstrated that miR-382 was downregulated in RB tissues and cell lines. Upregulation of miR-382 inhibited RB cell proliferation and invasion in vitro. Additionally, brain-derived neurotrophic factor (BDNF) was identified as a novel target of miR-382 in RB. BDNF was upregulated in RB tissues and negatively associated with miR-382 expression levels. Furthermore, BDNF overexpression rescued the tumour-suppressing effects on RB cells induced by miR-382. miR-382 inactivated the phosphoinositide 3-kinase/protein kinase B (PI3K/AKT) signalling pathway in RB. These findings suggested that miR-382 serves as a tumour suppressor in RB, in part, by targeting the BDNF-mediated PI3K/AKT signalling pathway. The results of the present study suggest a potential therapeutic strategy for treating RB patients in the future.
关键词： PI3K/AKT,microRNA-382,brain-derived neurotrophic factor,retinoblastoma
更新于2025-09-23 15:22:29
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An Enzyme-Free MicroRNA Assay Based-on Fluorescence Counting of Click Chemical Ligation-Illuminated Magnetic Nanoparticles with Total Internal Reflection Fluorescence Microscopy
摘要： MicroRNAs (miRNAs) have been considered as promising cancer biomarkers. However, the simple but sensitive detection of low levels of miRNAs in biological samples still remains challenging. Herein, we wish to report an entirely enzyme-free, simple and highly sensitive miRNA assay based on the counting of cycling click chemical ligation (3CL)-illuminated fluorescent magnetic nanoparticles (MNPs) with a total internal reflection fluorescence microscopy (TIRFM). In this strategy, each miRNA molecule can trigger many cycles of click chemical ligation reactions to produce plentiful ligated oligonucleotides (ODNs) with both 5’-biotin and 3’-fluorophore, resulting in efficient signal amplification. It is worth noting that only the ligated ODNs can bring fluorophores onto streptavidin-functionalized MNPs (STV-MNPs). Notably, merely 10 fluorescent molecules on each 50 nm MNP can make it bright enough to be clearly visualized by the TIRFM, which can significantly improve the detection sensitivity for miRNA. Through fluorescence counting of individual MNPs and integrating their fluorescence intensities, the amount of target miRNA can be quantitatively determined. This miRNA assay can be accomplished in a mix-and-read manner just by simply mixing the enzyme-free 3CL reaction system with the MNPs before TIRFM imaging, which avoids tedious immobilization, washing and purification steps. Despite the extremely simple operation, this strategy exhibits high sensitivity with a quite low detection limit of 50 fM target miRNA as well as high specificity to well discriminate miRNA sequences with a single-base variation. Furthermore, the applicability of this method in real biological samples is also verified through the accurate detection of miRNA target in cancer cells.
关键词： click chemistry,microRNA,TIRFM,magnetic nanoparticle,fluorescence counting
更新于2025-09-23 15:21:21
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MicroRNA-9 inhibits retinal neovascularization in rats with diabetic retinopathy by targeting vascular endothelial growth factor A
摘要： Diabetic retinopathy (DR) is a leading cause of adult visual impairment and loss. This study aims to explore the effects of microRNA‐9 (miR‐9) on retinal neovascularization during DR by targeting the vascular endothelial growth factor A (VEGFA). DR rat models were successfully established. Retinal microvascular endothelial cells (RMECs) of DR rats were isolated and treated with miR‐9 mimic, miR‐9 inhibitor or small interfering RNA (siRNA)‐VEGFA. The expressions of miR‐9, VEGFA, and cluster of differentiation 31 (CD31) of the rats’ tissues and cells were examined. The targeting relationship between miR‐9 and VEGFA was testified. The tubule formation, the cell proliferation and the periodic distribution and apoptosis were evaluated after transfection. In the retinal tissues of DR rats, miR‐9 expression decreased while the expression of VEGFA and CD31 increased. Notably, miR‐9 targeted and inhibited VEGFA expression. In response to the treatment of miR‐9 mimic and siRNA‐VEGFA, a reduction was identified in CD31 expression, tubule formation, and proliferation of RMECs and cell ratio in the S phase, but an increase was observed in apoptosis rate of RMECs. The treatment of miR‐9 inhibitor reversed the manifestations. Our study demonstrated that miR‐9 could inhibit retinal neovascularization of DR and tubule formation, and promote apoptosis in RMECs by targeting VEGFA.
关键词： retinal microvascular endothelial cells,diabetic retinopathy,vascular endothelial growth factor A,microRNA‐9,retinal neovascularization
更新于2025-09-23 15:21:21