Novel Cells-Based Electrochemical Sensor for Investigating the Interactions of Cancer Cells with Molecules and Screening Multitarget Anticancer Drugs.

A novel, effective, and label-free electrochemical sensor was constructed for investigating the interactions between cancer cells and molecules, based on targeted cancer cells immobilized on a bilayer architecture of N-doped graphene-Pt nanoparticles-chitosan (NGR-Pt-CS) and polyaniline (PANI). The interactions between folic acid (FA, positive control) and dimethyl sulfoxide (DMSO, negative control) and the choice of targeted cells, HepG2 and A549 cells, were investigated by measuring the current change of the sensor to [Fe(CN)6]3-/4- before and after interactions, and the binding constants were calculated to be 1.37 × 105 and 1.92 × 105 M-1 by sensing kinetics. Furthermore, 18 main components from Aidi injection (ADI) were studied to screen compounds that have interactions with different targeted cancer cells including HepG2 and A549 cells. The potential target groups of the interactions between screened active compounds and targeted cancer cells were analyzed through computer-aided molecular docking. In this sensing system, molecules did not require electrochemical activity, and different targeted cancer cells could be immobilized on the modified electrode surface, truly reflecting the categories and numbers of targets. Additionally, the proposed sensor specifically circumvented the current paradigm in most cells-based electrochemical sensors for screening drugs, in which the changes in cell behavior induced by drugs are monitored. This study provided a novel, simple, and generally applicable method for exploring the interaction of molecules with cancer cells and screening multitarget drugs.

Rapid screening of multi-target antitumor drugs by nonimmobilized tumor cells/tissues capillary electrophoresis.

As there are more target categories on tumor cells/tissues than on receptor-overexpressing cells, and tumor tissues can better simulate TME, we established a new method of screening multi-target antitumor drugs by nonimmobilized tumor cells/tissues capillary electrophoresis under approximately tumor physiological environment. In this method, the natural structure and active conformation of the target proteins on tumor cells/tissues can be well maintained without separation and purification. Therefore, we successfully used this method to study the interactions between the Aidi injection (ADI)/its main components and tumor cells/tissues by optimizing a series of experimental conditions, discovered seven components with binding activity to A549 cells, five of them with specific interaction to tumor tissues, and calculated the binding kinetic parameters (K, ka, kd, and k'). Then, antitumor activity assays in vitro and in vivo were carried out to discover a new drug combination with higher targeting, better pharmaceutical efficacy, and lower toxic side effects. Finally, molecular docking studies were performed to investigate the potential target groups of the interactions between the effective drug combination and A549 cells/tissues. In summary, the method was verified to be valid and feasible, and can be easily transferred to a capillary array electrophoresis for high-throughput drug screening.

Mechanisms of the active components from Korean pine nut preventing and treating d-galactose-induced aging rats.

Age-related neuronal injury and oxidative damage are the predominant factors for neurodegenerative diseases like Alzheimer's disease (AD). The aim of this study was to explore whether chronic administration of d-galactose (d-gal) can cause neuronal injury and oxidative damage, and to investigate the neuroprotective and antioxidative effects of the active components (UPNO-1) from Korean pine nut (Pinus koraiensis). Two dosing regimens were designed, one for the evaluation of preventive effects in which the rats were simultaneously administrated d-gal and UPNO-1/fishoil for 12 weeks, the other for the evaluation of therapeutic effects in which the rats were given d-gal for 8 weeks before treated with UPNO-1/selegiline for 8 weeks. The experimental results demonstrated that chronic administration of d-gal produced histopathological changes and increased neuronal apoptosis, and decreased significantly the activities of T-AOC, T-SOD and CAT. Additionally, a comprehensive metabolic profiling of d-gal-treated rats was performed for the first time to investigate the metabolic disorders in the hippocampus, cortex and plasma, and a total of 32 annotated metabolites were significantly increased or decreased in the modeled rats. Major disturbed metabolic pathways were fatty acid, glycerolphospholipid and arachidonic acid metabolic pathways. UPNO-1 significantly diminished neuronal apoptosis, ameliorated histopathological findings, and increased the activities of T-SOD and CAT but not T-AOC. Furthermore, UPNO-1 attenuated the decreased plasma levels of 3-oxooctanoic acid, l-tryptophan, 12-hydroxyheptadecanoic acid, lysophosphatidylcholine (16:0) (LPC(16:0)), LPC(18:3) and LPC(18:1) in the modeled rats. These results illustrated the mechanisms of d-gal induced neurotoxicity and oxidative stress and proved the positive effects of UPNO-1 on preventing and treating d-gal-induced-aging rats.

Nonimmobilized Biomaterial Capillary Electrophoresis for Screening Drugs Targeting Human Glucose Transporter 1.

We report an online ligand screening method that targets human glucose transporter 1 (hGlut1) under approximately physiological conditions, named nonimmobilized biomaterial capillary electrophoresis (NIBCE), and we investigated the interactions between drugs/candidate compounds and HEK293 cells, hGlut1-overexpressing HEK293 cells, non-small-cell lung cancer A549 cells, A549 tumor tissue, and normal lung tissue by simulating the interactions between drugs and moving target cells or the space-occupying tumor. NIBCE omits the trouble of isolating and purifying target receptors from cell membrane while maintaining their native conformation and binding activity. The biomaterials were intercepted by porous frits in capillary columns and cannot flow through the detection window, thereby solving the problem of interference detection, and they can be renewed any time flexibly, thus effectively maintaining their surface bioactivity. Furthermore, the binding kinetic parameters (K, ka, kd, and k') were calculated by nonlinear chromatography (NLC) theory, and competitive binding experiments, ligand docking studies, and antitumor activity assays in vitro and in vivo were performed to verify the feasibility of NIBCE.