Presence Of Mucin 1 Research Articles

Dual electrochemical signal “signal-on-off” sensor based on CHA-Td-HCR and CRISPR-Cas12a for MUC1 detection

Mucin 1 (MUC1) is frequently overexpressed in various cancers and is essential for early cancer detection. Current methods to detect MUC1 are expensive, time-consuming, and require skilled personnel. Therefore, developing a simple, sensitive, highly selective MUC1 detection sensor is necessary. In this study, we proposed a novel “signal-on-off” strategy that, in the presence of MUC1, synergistically integrates catalytic hairpin assembly (CHA) with DNA tetrahedron (Td)-based nonlinear hybridization chain reaction (HCR) to enhance the immobilization of electrochemically active methylene blue (MB) on magnetic nanoparticles (MNP), marking the MB signal “on”. Concurrently, the activation of CRISPR-Cas12a by isothermal amplification products triggers the cleavage of single-stranded DNA (ssDNA) at the electrode surface, resulting in a reduction of MgAl-LDH@Fc-AuFe-MIL-101 (containing ferrocene, Fc) on the electrode, presenting the “signal-off” state. Both MB and MgAl-LDH@Fc-AuFe-MIL-101 electrochemical signals were measured and analyzed. Assay parameters were optimized, and sensitivity, stability, and linear range were assessed. Across a concentration spectrum of MUC1 spanning from 10 fg/mL to 100 ng/mL, the MB and MgAl-LDH@Fc-AuFe-MIL-101 signals were calibrated with each other, demonstrating a “signal-on-off” dual electrochemical signaling pattern. This allows for the precise and quantitative detection of MUC1 in clinical samples, offering significant potential for medical diagnosis.

Colorimetric aptasensor based on magnetic beads and gold nanoparticles for detecting mucin 1

In this work, a colorimetric aptasensor based on magnetic beads (MBs), gold nanoparticles (AuNPs) and Horseradish Peroxidase (HRP) was prepared for the detection of mucin 1 (MUC1). Complementary DNA of the MUC1 aptamer (Apt) immobilized on the MBs was combined with the prepared AuNPs-Apt-HRP complex (AuNPs@Apt-HRP). In the presence of MUC1, it specifically bound to Apt, resulting in the detachment of gold nanoparticles from the MBs. After magnetic separation, AuNPs@Apt-HRP was separated into the supernatant and reacted with 3,3′,5,5′-Tetramethylbenzidine (TMB) to produce color reaction from colorless to blue. The linear range of MUC1 was from 75 to 500 μg/mL (R2 = 0.9878), and the detection limit was 41.95 μg/mL. The recovery rate of MUC1 in human serum was 99.18 %∼101.15 %. This method is simple and convenient. Moreover, it does not require complex and expensive equipment for detection of MUC1. It provides value for the development of MUC1 colorimetric sensors and a promising strategy for the determination of MUC1 in clinical diagnosis.

The novel few layered MXene/MoS2/PdPtNPs nanomaterials combined with hybrid chain reaction for electrochemical detection of mucin 1

This work has developed a novel and highly sensitive electrochemical aptasensor for the detection of mucin 1 (MUC1) based on the MXene/MoS2/PdPtNPs nanomaterials and hybridization chain reaction (HCR). MXene/MoS2/PdPtNPs nanomaterials were first prepared via a simple hydrothermal and external reduction method, which was used as the supporting substrate of the biosensor. MXene synergized with MoS2-PdPtNPs could prevent the few layered MXene from curling effectively. Besides, the proposed sensing platform could simultaneously increase the active site and enhance the electrochemical performance. In the presence of MUC1, dsDNA structural switching was induced by MUC1 binding to aptamer specifically, and most aptamers were pulled off from the electrode surface, which thus resulted in the assembly of initiator S, harpin probe 1 (H1) and harpin probe 2 (H2) on electrode surface to trigger the HCR. Since the peroxidase-like mimic manganese (III) meso-tetrakis (4-Nmethylpyridyl)-porphyrin (MnTMPyP) interacts with DNA nanoladders via groove binding, the combined PdPtNPs could result in a great current response to the hydrogen peroxide hydroquinone system. For MUC1 detection, the proposed biosensor has showed a wide linearity range of 1 pg mL−1–10 ng mL−1 and a low detection limit of 0.36 pg mL−1 (S/N = 3), which was hopeful to be applied in biotechnology and clinical diagnosis.

Irradiation Alters the Expression of MUC1, CD44 and Hyaluronan in Oral Mucosal Epithelium.

Purpose: It is well established that cancer cells exploit aberrant synthesis of mucin 1 (MUC1) and hyaluronan (HA) synthesis along with HA's physiological cell surface receptor CD44. However, their role in irradiated oral tissue has not been reported previously. We, therefore, aimed to study MUC1, CD44 and HA immunohistochemically in irradiated oral mucosa and their role in the long-term effects after radiotherapy. Materials and Methods: Oral mucosal biopsies were obtained from healthy subjects as controls and from patients after radiotherapy for head and neck cancer (irradiated group) during dental implant surgery.The presence of MUC1, CD44, and HA in oral mucosa was studied by immunohistochemical methods. The differences in the localization and intensity in the oral epithelium between control and irradiated tissue were analyzed. Results: The staining intensity of MUC1 was confined to the superficial epithelial layer, whereas HA and CD44 were found in the cell membranes in the epithelial basal and intermediate layers of control specimens. In irradiated epithelium, MUC1 staining was distributed throughout all the layers of the oral epithelium, with significant staining in the basal and intermediate layers. Accordingly, HA and CD44 staining extended to involve the superficial cells of the irradiated epithelium. The staining pattern of MUC1 and CD44 showed significant changes in irradiated samples. Conclusions: Our results showed that the staining intensities of MUC1, CD44, and HA were significantly elevated in irradiated tissue compared to controls. MUC1, CD44, and HA are important markers and take part in long-term changes in the oral mucosa after radiotherapy.

A portable multicolor aptasensor for MUC1 detection based on enzyme-mediated cascade reaction

Mucin 1 (MUC1) is an important tumor biomarker, and the determination of MUC1 is of great significance in clinical diagnostics, prognosis assessment and therapy monitoring. Developing point-of-care testing method of MUC1 is necessary in resource-limited settings. Herein, a visual multicolor aptasensor for MUC1 detection was designed based on an alkaline phosphatase (ALP)-triggered cascade reaction. In the presence of MUC1, ALP-decorated complementary DNA and MUC1 aptamer on magnetic beads were dissociated due to the recognition between MUC1 and aptamer. With the assistance of a magnet, ALP-decorated complementary DNA was separated and collected to catalyze the hydrolysis of l-ascorbic acid-2-phosphate generating l-ascorbic acid, which etched the manganese dioxide nanosheets to mediate the oxidation reaction of 3,3′,5,5′-tetramethylbenzidine sulfate, resulting in different products with distinct visible multicolor. The naked eye detection of MUC1 was realized and the detection limit was estimated to be 0.18 U mL−1 by using the colorimetric method. This proposed aptasensor possesses the merit of vivid multicolor change, and provides a new way for MUC1 detection by naked eyes.

Highly sensitive detection of MUC1 by microchip electrophoresis combining with target recycling amplification and strand displacement amplification

Mucin 1 (MUC1) is usually overexpressed in a variety of malignant tumors, and quantitative analysis of MUC1 plays an important role in the early diagnosis of cancer. In this work, a highly sensitive MUC1 assay was developed by integrating microchip electrophoresis (MCE) with target recycling amplification (TRA) and strand displacement amplification (SDA). Specifically, the presence of MUC1 can trigger the exposure of the designed hairpin probe (HP) to initiate SDA and an amplified amount of ssDNA is produced finally. The amount of these ssDNA can be detected by MCE, then the concentration of MUC1 can be obtained through the correlation between MUC1 concentration and ssDNA concentration. The experimental results show that the MCE signal had a good linear relationship with MUC1 concentration in the range of 1.0 pg/mL - 1.0 × 103 pg/mL with a low limit of detection of 0.23 pg/mL under the optimal conditions (S/N = 3). Additionally, the assay had been successfully applied to detect MUC1 in biological samples with satisfactory results, providing an alternative assay for the detection of other tumor markers owing to the high sensitivity, high selectivity, simple operation and low sample consumption.

Tetraphenylethylene-Functionalized Metal-Organic Frameworks with Strong Aggregation-Induced Electrochemiluminescence for Ultrasensitive Analysis through a Multiple Convertible Resonance Energy Transfer System.

Since aggregation-induced electrochemiluminescence (AIECL) combined the merits of aggregation-induced emission (AIE) and electrochemiluminescence (ECL), it has become a research hotspot recently. Herein, novel kinds of functional metal-organic frameworks (MOFs) with strong AIECL were reported through doping tetraphenylethylene (TPE) into UiO-66. Due to the porosity and highly ordered topological structure that caused the confinement effect of MOFs, the molecular motion of TPE was effectively limited within UiO-66, resulting in strong AIE. Meanwhile, the large specific surface area and porous structure of UiO-66 allowed TPE to react with coreactants more effectively, which was beneficial to ECL. Thus, the TPE-functionalized UiO-66 (TPE-UiO-66) showed excellent AIECL performance surprisingly. Inspired by this, a multiple convertible ECL resonance energy transfer (ECL-RET) system was constructed through a DNA Y structure that regulated the distance between the energy donor (TPE-UiO-66) and different energy acceptors (gold nanoparticles and Adriamycin). Furthermore, an ultrasensitive ECL biosensor for the detection of Mucin 1 (MUC1) was developed through the introduction of the novel ECL-RET system. In the presence of MUC1, the DNA Y structure was constructed, keeping the gold nanoparticles (AuNPs) away from TPE-UiO-66. Then, Adriamycin (Dox) could be embedded in the DNA Y structure and act as an energy acceptor to receive the energy of TPE-UiO-66, which made the biosensor produce a strong ECL response. As expected, the developed ECL biosensor exhibited superior detection performance for MUC1. This work provided a novel way to realize AIECL and board the application of AIECL in analytical chemistry.

MUC1 detection and in situ imaging method based on aptamer conformational switch and hybridization chain reaction

Mucin 1 (MUC1) overexpression in tumor cells is related to various cancers, including breast, stomach, and lung cancer. MUC1 detection and imaging are important for cancer localization in tissue sections to support histopathological diagnosis. In this study, we developed a simple, enzyme-free MUC1 detection and in situ imaging method. Three hairpin probes, Apt-trigger, HP1-FAM, and HP2, were designed for MUC1 recognition and hybridization chain reaction (HCR). The Apt-trigger probe was composed of two sequences: the MUC1 aptamer and HCR trigger sequence. The 5’ end of the HP1-FAM probe was modified with a FAM signal molecule. In the presence of MUC1, the aptamer sequence is activated and bound to MUC1, which opens the hairpin structure. Then, the trigger sequence gets exposed and, complementary to HP1-FAM, triggers a continuous HCR process. This method was successfully used to detect MUC1 of 200 pM–25 nM and MUC1 in situ imaging in specific cells, such as human breast carcinoma (MCF-7) and human colon cancer (HT-29) cells.

A photocurrent-polarity-switching biosensor for highly selective assay of mucin 1 based on target-induced hemin transfer from ZrO2 hollow spheres to G-quadruplex nanowires

Herein, a photocurrent polarity switching platform for highly selective assay of mucin 1 (MUC1) was developed based on target-induced hemin transfer from ZrO2 hollow spheres (ZrO2 HSs) to G-quadruplex nanowires (G wires). In this system, SiO2 spheres were used as templates to synthesize the uniform and mesoporous ZrO2 HSs. As nanocontainers, ZrO2 HSs could load hemin in its cavity via pores. Then, the aptamers of MUC1, as bio-gates, blocked the pores of ZrO2 HSs based on the specific binding of Zr4+ and the phosphate groups of aptamer. In the presence of MUC1, the aptamer could specifically recognize and bind with MUC1, and then leave away from the surface of ZrO2 HSs, which resulted in the opening of the bio-gates and releasing of hemin. Assisted with the G wires formed on the Au NPs/In2S3/ITO, the released hemin was captured on the electrode through the formation of hemin/G-quadruplex structure, leading to the switch of the photocurrent polarity of the electrode from anodic photocurrent to cathodic photocurrent. The proposed photoelectrochemical biosensor showed outstanding performance for MUC1 assay with high selectivity, wide linear response range (1 fg mL−1 -10 ng mL−1) and lower detection limit (0.48 fg mL−1). And the strategy could be easily extended to a triple-mode detection of MUC1 because the hemin/G-quadruplex structure was widely used in electrochemical and colorimetric methods as a hydrogen peroxide mimetic enzyme, which might provide wide applications in biological or clinical studies.

Multiple signal amplification via coupling DNAzyme with strand displacement reaction for sensitive colorimetric analysis of MUC1

Mucin 1 (MUC1) is abnormally expressed in tumor tissues, and quantitative analysis of MUC1 is thus beneficial to the prevention and early diagnosis of cancer. In this work, by intergrating DNAzyme with strand displacement reaction (SDR), we have fabricated a highly sensitive MUC1 colorimetric assay based on a multiple signal amplification strategy. Specifically, the presence of MUC1 can induce the exposure of the toehold region pre-locked in the aptamer probe (AP) to initiate SDR, liberating the MUC1/AP complex for recycling (cycle I) and forming the dimer-like DNAzyme. The formed DNAzyme can then cyclically cleave the substrate signal probe (cycle II), triggering the formation of G-quadruplex sequence and the new active enzyme sequence that can also cleave the signal probe (cycle III). Such target-induced triple amplification reaction can generate massive G-quadruplex sequences, which leads to a dramatically enhanced colorimetric signal for sensitive detection of MUC1 even at an ultra-low level of 35 pM. The colorimetric strategy also holds a high selectivity to distinguish MUC1 from other interfering proteins and can be successfully applied to the detection of MUC1 in human serum as well as cancer cells, presenting great potential in biosensing and clinical diagnostics.

Aptamer-functionalized polydiacetylene liposomes act as a fluorescent sensor for sensitive detection of MUC1 and targeted imaging of cancer cells

Mucin 1 (MUC1) is an important biomarker that has been shown to participate in the metastasis and invasion of various types of cancers. Simple yet sensitive methods for the detection of MUC1 in liquid and in situ imaging of MUC1 at cellular level are crucial for early diagnosis of cancers as well as monitoring their progression. Herein, a polydiacetylene (PDA) liposome-based sensing system functionalized with Cy3-labeled MUC1 binding aptamer (Cy3-Apt) was developed as a fluorescence “turn-on” nanosensor for the detection of MUC1. In the sensing system, the fluorescence of Cy3-Apt could be directly quenched due to the energy transfer between the fluorogen and PDA conjugated backbone. In the presence of MUC1, the recognition of MUC1 with its aptamer induces the structure switching of both PDA liposomes and the aptamer, recovering the red fluorescence. Sensitive and selective analysis of MUC1 in aqueous media could be achieved with a limit of detection around 0.8 nM. The feasibility of the nanosensor was further demonstrated by mapping the spatial expression of MUC1 in cancer cells with excellent sensitivity.

Enhanced photoelectrochemical sensing for MUC1 detection based on TiO2/CdS:Eu/CdS cosensitized structure

A novel ultrasensitive photoelectrochemical aptasensing was developed to detect mucin 1 (MUC1) based on TiO2/CdS:Eu nanocomposite. The TiO2/CdS:Eu nanocomposite modified fluorine-doped tin oxide (FTO) electrode was prepared by successive ionic layer adsorption and reaction technique (SILAR) and then was employed as matrix for immobilization of SH-DNA (DNA1). After 6-mercaptohexanol (MCH) blocked unbound sites of the electrode surface, the MUC1 aptamer(DNA2) was introduced, allowing MUC1 aptamer to hybridize with DNA1. Subsequently, CdS-COOH QDs as signal amplification elements were successively labeled on the terminal of MUC1 aptamer. In this case, the target MUC1 detection was based upon the photocurrent change originated from quantities change of the signal amplification elements CdS. In the presence of MUC1, the MUC1 aptamer was specifically bound with MUC1, and meanwhile MUC1/DNA2/CdS was released from the electrode surface, resulting in significantly decreased photocurrent intensity. The novel ultrasensitive photoelectrochemical aptasensing presented a highly selective, sensitive and reproductive detection of MUC1 from 1 nM to 5 μM with a lower detection limit of 0.3 nM. At the same time, it also provided a good specificity, reproducibility, stability and promising strategy to detect a variety of other protein targets for early clinical diagnosis of different diseases.

Mucin 1, a signal transduction membrane-bound mucin, is present in human disc tissue and is downregulated in vitro by exposure to IL-1\xdf or TNF-\u03b1

BackgroundBack pain and disc degeneration have a growing socioeconomic healthcare impact. Mucin 1 (MUC1) is a transmembrane glycoprotein whose extracellular and intracellular domains participate in cellular signaling. Little is currently known about the presence or role of MUC1 in human disc degeneration.MethodsIn this IRB-approved research study, 29 human disc specimens were analyzed for MUC1 immunohistochemical localization and gene expression, and annulus fibrosus (annulus) cells were also isolated and cultured in 3D. Microarray analysis assessed expression levels of MUC1 in healthy and degenerated disc tissue and in cells exposed to proinflammatory cytokines (IL-1ß or TNF-α).ResultsMUC1 was shown to be present in annulus cells at the protein level using immunochemistry, and its expression was significantly upregulated in annulus tissue from more degenerated grade V discs compared to healthier grade I-II discs (p = 0.02). A significant positive correlation was present between the percentage of MUC1-positive cells and disc grade (p = 0.009). MUC1 expression in annulus cells cultured in 3D was also analyzed following exposure to IL-1ß or TNF-α; exposure produced significant MUC1 downregulation (p = 0.0006).ConclusionsHere we present the first data for the constitutive presence of MUC1 in the human disc, and its altered expression during disc degeneration. MUC1 may have an important role in disc aging and degeneration by acting as a regulator in the hypoxic environment, helping disc cells to survive under hypoxic conditions by stabilization and by activation of HIF-1α as previously recognized in pancreatic cancer cells.

SERS- and luminescence-active Au-Au-UCNP trimers for attomolar detection of two cancer biomarkers.

Alpha-fetoprotein (AFP) and mucoprotein1 (mucin-1) are two important disease biomarkers. Self-assembled gold nanoparticles (AuNPs) and upconversion nanoparticle (Au-Au-UCNP) trimers based on aptamers were developed for the ultrasensitive detection of AFP and mucin-1. The Au-Au-UCNP trimers produced ideal optical signals, with prominent Raman enhancement and fluorescence quenching effects. The surface-enhanced Raman scattering (SERS) intensity decreased in the presence of mucin-1 and the luminescence intensity increased in the presence of AFP. A limit of detection (LOD) of 4.1 aM and a wide linear range of 0.01-10 fM for the detection of mucin-1 were obtained with this SERS-encoded sensing system. Using the luminescence-encoded sensing system, a LOD of 0.059 aM and a wide linear range of 1-100 aM for the detection of AFP were obtained. These LODs are the lowest values reported so far. This approach has the advantage of detecting two disease biomarkers simultaneously.

Fluorescence aggregation assay for the protein biomarker mucin 1 using carbon dot-labeled antibodies and aptamers

We have developed a sensitive fluorescence assay for the protein biomarker mucin 1 (MUC1). It is based on the aggregation of functionalized carbon dots (CDs) in the presence of MUC1. The CDs with a size of ~2 nm display strong blue-green intrinsic fluorescence. Antibodies against MUC1 and the MUC1 aptamer were covalently conjugated to the CDs, and the immunoreaction between CD-labeled antibody to MUC1, and of the CD-labeled aptamer to MUC1 resulted in the formation of a sandwich structure that is accompanied by aggregation of CDs and quenching of fluorescence. The change in fluorescence is linearly related to the MUC1 concentration in the 5 to 100 nM range, with a detection limit at 2 nM. The assay was applied to the determination of MUC1 in (spiked) real serum samples, and good recoveries (ranging from 94.2 to 98.1 %) were obtained. This assay is cost-effective and convenient. It has a wide scope in that it, conceivably, can be extended to assays for other proteins and biomarkers of clinical interest.

Expression of MUC1 mucin in human umbilical vein endothelial cells (HUVEC).

Mucin 1 (MUC1) is a membrane-bound glycoprotein that is expressed by various epithelial cell types. MUC1 functions include modulation of cell adhesion, signal transduction, lubrication and hydration of epithelial surfaces, and their protection from infection. In this study we demonstrated that MUC1 is expressed in human umbilical vein endothelial cells (HUVECs) and could be released/shed from cellular membrane. MUC1 presence in these cells was verified using three methods: Western blotting, flow cytometry and metabolic labeling. We also showed that mucin expression is stimulated by proinflammatory cytokines: about a 2-fold increase was observed after TNF-α treatment and lower after IFN-γ alone and in combination with TNF-α treatment. It can be assumed that the presence of MUC1 in endothelial cells may have an important role in the interactions with different cell types in physiological and pathological processes.

Evaluation of ocular surface disorders: a new diagnostic tool based on impression cytology and confocal laser scanning microscopy

AimTo provide a new tool for the evaluation of altered ocular surfaces by using a combination of impression cytology, laser scanning confocal microscopy and advanced image analysis.MethodsThe expression of keratin...