Detection of embryonic stem cell lysate biomarkers by surface plasmon resonance with reduced nonspecific adsorption

Tyagi, Deependra; Pérez, Javier Batista; Nand, Amita; Cheng, Zhiqiang; Wang, Peizhe; Na, Jie; Zhu, Jianming

doi:10.1016/j.ab.2014.11.001

Cited by 17 publications

(7 citation statements)

References 45 publications

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“…As the most commonly used clinical anti-glioma agent, TMZ has the issues of increased resistance and various side effects for GBM patients. 28,29 Thus, an auxiliary drug that can enhance the therapeutic effect of TMZ or reduce the dosage of TMZ will substantially improve the prognosis of GBM patients. In the current study, we further explored the anti-glioma mechanisms of CS-6 and, more importantly, clarified its chemosensitizing effect on TMZ, which will provide a theoretical basis for future clinical research on CS-6.…”

Section: Discussionmentioning

confidence: 99%

“…Our previous research suggested that bufadienolides could inhibit GBM via promoting ATP1A1 degradation. 28 Thus, we examined whether ATP1A1 impacts the synergistic effect of TMZ and CS-6. We knocked down ATP1A1 in U87 cells by using an shRNA strategy (Figure 1H); however, suppressing ATP1A1 in U87 cells did not affect the synergistic therapeutic effect of CS-6 and TMZ (Figure 1I).…”

Section: Cs-6 Improves Tmz Sensitivity In Glioma Cells Regardless Of ...mentioning

confidence: 99%

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Gamabufotalin induces a negative feedback loop connecting ATP1A3 expression and the AQP4 pathway to promote temozolomide sensitivity in glioblastoma cells by targeting the amino acid Thr794

et al. 2019

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Objectives: Temozolomide (TMZ) is one of the most commonly used clinical drugs for glioblastoma (GBM) treatment, but its drug sensitivity needs to be improved.Gamabufotalin (CS-6), the primary component of the traditional Chinese medicine "ChanSu," was shown to have strong anti-cancer activity. However, more efforts should be directed towards reducing its toxicity or effective treatment doses.Methods: Target fishing experiment, Western blotting, PCR, confocal immunofluorescence and molecular cloning techniques were performed to search for possible downstream signalling pathways. In addition, GBM xenografts were used to further determine the potential molecular mechanisms of the synergistic effects of CS-6 and TMZ in vivo. Results: Mechanistic research revealed a negative feedback loop between ATP1A3and AQP4 through which CS-6 inhibited GBM growth and mediated the synergistic treatment effect of CS-6 and TMZ. In addition, by mutating potential amino acid residues of ATP1A3, which were predicted by modelling and docking to interact with CS-6, we demonstrated that abrogating hydrogen bonding of the amino acid Thr794 interferes with the activation of ATP1A3 by CS-6 and that the Thr794Ala mutation directly affects the synergistic treatment efficacy of CS-6 and TMZ. Conclusions:As the main potential target of CS-6, ATP1A3 activation critically depends on the hydrogen bonding of Thr794 with CS-6. The combination of CS-6 and TMZ could significantly reduce the therapeutic doses and promote the anti-cancer efficacy of CS-6/TMZ monotherapy.

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Section: Discussionmentioning

confidence: 99%

Section: Cs-6 Improves Tmz Sensitivity In Glioma Cells Regardless Of ...mentioning

confidence: 99%

Gamabufotalin induces a negative feedback loop connecting ATP1A3 expression and the AQP4 pathway to promote temozolomide sensitivity in glioblastoma cells by targeting the amino acid Thr794

et al. 2019

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“…Poly (ethylene glycol) (PEG) and its derivatives are the most commonly used anti-fouling materials for biosensor construction. For example, in a study used for detecting embryonic stem cell lysate with surface plasmon resonance imaging, the surface was modified with thiolated PEG chains or three dimensional polymers containing PEG chains [ 18 ]. These surfaces combined with a good blocking solution, such as amine terminated PEG, and adequate running buffer supplemented with detergent exhibited greatly decreased non-specific binding.…”

Section: Introductionmentioning

confidence: 99%

Optimisation and Characterisation of Anti-Fouling Ternary SAM Layers for Impedance-Based Aptasensors

Miodek

Regan

Bhalla

et al. 2015

Sensors

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An aptasensor with enhanced anti-fouling properties has been developed. As a case study, the aptasensor was designed with specificity for human thrombin. The sensing platform was developed on screen printed electrodes and is composed of a self-assembled monolayer made from a ternary mixture of 15-base thiolated DNA aptamers specific for human thrombin co-immobilised with 1,6-hexanedithiol (HDT) and further passivated with 1-mercapto-6-hexanol (MCH). HDT binds to the surface by two of its thiol groups forming alkyl chain bridges and this architecture protects from non-specific attachment of molecules to the electrode surface. Using Electrochemical Impedance Spectroscopy (EIS), the aptasensor is able to detect human thrombin as variations in charge transfer resistance (Rct) upon protein binding. After exposure to a high concentration of non-specific Bovine Serum Albumin (BSA) solution, no changes in the Rct value were observed, highlighting the bio-fouling resistance of the surface generated. In this paper, we present the optimisation and characterisation of the aptasensor based on the ternary self-assembled monolayer (SAM) layer. We show that anti-fouling properties depend on the type of gold surface used for biosensor construction, which was also confirmed by contact angle measurements. We further studied the ratio between aptamers and HDT, which can determine the specificity and selectivity of the sensing layer. We also report the influence of buffer pH and temperature used for incubation of electrodes with proteins on detection and anti-fouling properties. Finally, the stability of the aptasensor was studied by storage of modified electrodes for up to 28 days in different buffers and atmospheric conditions. Aptasensors based on ternary SAM layers are highly promising for clinical applications for detection of a range of proteins in real biological samples.

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“…However, these strategies are still unable to identify direct interactions between protein targets and drug molecules. Surface plasmon resonance (SPR) biosensor technology has been widely used to characterize unmodified biological drugs and identify the macromolecular targets of small molecular agents 24 , 25 . The advantages of this approach are that there is no need for the labeling and real time monitoring of intermolecular interaction.…”

Section: Introductionmentioning

confidence: 99%

TRPM8-regulated calcium mobilization plays a critical role in synergistic chemosensitization of Borneol on Doxorubicin

Lai¹,

Liu²,

Hou³

et al. 2020

Theranostics

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Background: Lung cancer has a high mortality rate and is resistant to multiple chemotherapeutics. Natural Borneol (NB) is a monoterpenoid compound that facilitates the bioavailability of drugs. In this study, we investigated the effects of NB on chemosensitivity in the A549 human lung adenocarcinoma cell line and to elucidate therapeutic molecular target of NB. Methods: The chemosensitivity effects of NB in A549 cells were examined by MTT assay. The mechanism of NB action was evaluated using flow cytometry and Western blotting assays. Surface plasmon resonance (SPR) and LC-MS combined analysis (MS-SPRi) was performed to elucidate the candidate molecular target of NB. The chemosensitizing capacity of NB in vivo was assessed in nude mice bearing A549 tumors. Results: NB pretreatment sensitized A549 cells to low doxorubicin (DOX) dosage, leading to a 15.7% to 41.5% increase in apoptosis. This increase was correlated with ERK and AKT inactivation and activation of phospho-p38 MAPK, phospho-JNK, and phosphor-p53. Furthermore, this synergism depends on reactive oxygen species (ROS) generation. MS-SPRi analysis revealed that transient receptor potential melastatin-8 (TRPM8) is the candidate target of NB in potentiating DOX killing potency. Genetically, TRPM8 knock-down significantly suppresses the chemosensitizing effects of NB and inhibits ROS generation through restraining calcium mobilization. Moreover, pretreatment with NB synergistically enhances the anticancer effects of DOX to delay tumor progression in vivo . Conclusions: These results suggest that TRPM8 may be a valid therapeutic target in the potential application of NB, and show that NB is a chemosensitizer for lung cancer treatment.

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Detection of embryonic stem cell lysate biomarkers by surface plasmon resonance with reduced nonspecific adsorption

Cited by 17 publications

References 45 publications

Gamabufotalin induces a negative feedback loop connecting ATP1A3 expression and the AQP4 pathway to promote temozolomide sensitivity in glioblastoma cells by targeting the amino acid Thr794

Gamabufotalin induces a negative feedback loop connecting ATP1A3 expression and the AQP4 pathway to promote temozolomide sensitivity in glioblastoma cells by targeting the amino acid Thr794

Optimisation and Characterisation of Anti-Fouling Ternary SAM Layers for Impedance-Based Aptasensors

TRPM8-regulated calcium mobilization plays a critical role in synergistic chemosensitization of Borneol on Doxorubicin

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