Tumor cell membrane-coated continuous electrochemical sensor for GLUT1 inhibitor screening / 药物分析学报

Glucose transporter 1(GLUT1)overexpression in tumor cells is a potential target for drug therapy,but few studies have reported screening GLUT1 inhibitors from natural or synthetic compounds.With cur-rent analysis techniques,it is difficult to accurately monitor the GLUT1 inhibitory effect of drug molecules in real-time.We developed a cell membrane-based glucose sensor(CMGS)that integrated a hydrogel electrode with tumor cell membranes to monitor GLUT1 transmembrane transport and screen for GLUT1 inhibitors in traditional Chinese medicines(TCMs).CMGS is compatible with cell membranes of various origins,including different types of tumors and cell lines with GLUT1 expression knocked down by small interfering RNA or small molecules.Based on CMGS continuous monitoring technique,we inves-tigated the glucose transport kinetics of cell membranes with varying levels of GLUT1 expression.We used CMGS to determine the GLUT1-inhibitory effects of drug monomers with similar structures from Scutellaria baicalensis and catechins families.Results were consistent with those of the cellular glucose uptake test and molecular-docking simulation.CMGS could accurately screen drug molecules in TCMs that inhibit GLUT1,providing a new strategy for studying transmembrane protein-receptor interactions.

Advances in research and application of breast cancer organoids / 中国组织工程研究

BACKGROUND: Breast cancer is one of the most common malignant tumors in women. Its incidence rate is increasing year by year and tends to be younger. It seriously threatens women’s health. Therefore, it is particularly important to establish an ideal breast cancer model that can accurately simulate the tumor in vivo. Organoid is a new three-dimensional cultural model in vitro, which recapitulates key aspects of in vivo tissue or organ. In recent years, researches based on organoids have covered many kinds of tumors. OBJECTIVE: To review the research progress and application of breast cancer organoids, in order to provide a new research way for personalized treatment of breast cancer. METHODS: Using the key words of “organoid, breast cancer organoids, cancer organoids, mammosphere, three-dimensional culture” in English and Chinese, respectively, the first author retrieved relevant articles published from January 1980 to February 2020 in CNKI, Wanfang, and PubMed databases. The type of the article was not limited. After removal of the articles that were not related to the purpose of the study or repetitive, 66 articles were finally analyzed. RESULTS AND CONCLUSION: This review introduced organoid technology briefly and retraced the process of exploring suitable culture conditions to establish breast-cancer-origin organoids. Also, we concluded latest development of its applications and research progress. Breast cancer organoids have a wide range of application prospects in disease modeling, tumor pathogenesis, drug screening and other aspects, which provide a reliable model for breast cancer research and treatment, and in particular, open up a new perspective for personalized treatment of breast cancer.

Selective and sensitive fluorescence imaging reveals microenvironment-dependent behavior of NO modulators in the endothelial system / 药物分析学报

Nitric oxide (NO) is a second messenger playing crucial roles in the signaling of a variety of cellular functions. Due to its pathophysiological significance, various NO modulators have been developed to explore NO pathways and some have been used as therapies. These modulators are often used directly to observe pharmacological effects in cell lines, but their actual effect on intracellular NO level is seldom analyzed. Herein, facilitated by a selective and sensitive fluorescence probe, we observed that some NO modulators displayed unexpected behaviors with both NO scavenger carboxy-PTIO and endothelial nitric oxide synthase (eNOS) inhibitor N(ω)-nitro-L-arginine methyl ester (L-NAME) failing to decrease intra-cellular free NO level in EA. hy926 cells while NO donor diethylamine-NONOate (DEA?NONOate) and eNOS activator calcimycin (A23187) failing to increase free NO level in human umbilical vein endothelial cell line (HUV-EC-C), although the reagents were confirmed to work normally in the primary human umbilical vein endothelial cells (primary HUVECs) and RAW 264.7 macrophage cells. Further research suggested that these unusual behaviors might be attributed to the cellular microenvironments including both the NO synthase (NOS) level and the endogenous glutathione (GSH) level. Genetically manipulating eNOS level in both cells restores the expected response, while decreasing GSH level restores the ability of DEA?NONOate to increase NO level in HUV-EC-C. These results reveal that the cellular microenvironment has a profound impact on pharmacological effect. Our study suggests GSH as a reservoir for NO in live cells and highlights the value of chemical probes as valuable tools to reveal microenvironment-dependent pharmacological effects.

3D-printing microfludic device for breast cancer cell culture and pharmacological research / 中国药学杂志

OBJECTIVE: To develop a microfluidic mammalian cell culture device with human breast cancer cells co-cultured with extracellular matrix (ECM), reconstitute the 3-dimension human body microenvironment and preliminarily conduct drug screening on the breast cancer chip. METHODS: 3D Printing technology was utilized to build a two-layer microchip with cell culture reservoirs. The breast cancer cells (MCF7) were cultured in the matrix gel structure which mimiced the 3-dimension human body microenvironment. The cell toxicity of paclitaxel (PTX) on MCF7 cells was preliminarily observed using this microfluidic chip. With the use of AO/EB immunofluorescent staining and laser confocal microscopy, the cell death ratio induced by PTX was determined and compared with that determined by 2-dimension drug screening methods. RESULTS: MCF7 cells cultured on the chip successfully formed a 3D cavity structure in the matrix after 6-day dynamic incubation. The flow rate was 10 μL · h-1. After 24 h dynamic culture, the culture medium was changed to culture solution containing 0.02 μmol · mL-1 PTX, and the incubation was continued for 24 h. Obvious cell death was detected under laser confocal microscopy. CONCLUSION: The microfluidic chip developed in this study can successfully culture breast cancer cells in 3-dimension structure and perform drug screening, which lays a foundation for actualization of "human-on-a-chip".

Amyotrophic Lateral Sclerosis - Cell Based Therapy and Novel Therapeutic Development

Amyotrophic lateral sclerosis (ALS) is a neurodegenerative disease, characterized by the predominant loss of motor neurons (MNs) in primary motor cortex, the brainstem, and the spinal cord, causing premature death in most cases. Minimal delay of pathological development by available medicine has prompted the search for novel therapeutic treatments to cure ALS. Cell-based therapy has been proposed as an ultimate source for regeneration of MNs. Recent completion of non-autologous fetal spinal stem cell transplant to ALS patients brought renewed hope for further human trials to cure the disease. Autologous somatic stem cell-based human trials are now in track to reveal the outcome of the ongoing trials. Furthermore, induced pluripotent stem cell (iPSC)-based ALS disease drug screen and autologous cell transplant options will broaden therapeutic options. In this review paper, we discuss recent accomplishments in cell transplant treatment for ALS and future options with iPSC technology.

Correlative Effect between in vivo Hollow Fiber Assay and Xenografts Assay in Drug Screening / Journal of the Korean Cancer Association, 대한암학회지

PURPOSE: This study was carried out to assess the usage of an in vivo hollow fiber assay to screen drugs with highly predictive accuracy. MATERIALS AND METHODS: The assay systems used were the hollow fiber and xenografts assays. The hollow fiber assay was carried out with the following steps; preparation of fibers, preparation of cells, loading and implanting fibers, treatment with drugs, removal of fibers and assaying for the cell viability by the MTT assay. For the xenografts assay, cell suspensions were subcutaneously transplanted into the mice. Therapy was started when the tumor volume reached 100~200 mm3. The tumor volumes were calculated using the formula V=[length+(width)2]/2, and used for evaluating the efficacy of the drugs. The drug treatment doses used were adriamycin 2.1 mg/kg, mitomycin-C 0.25 mg/kg, 5-fluo-rouracil 24.5 mg/kg and paclitaxel 2.5 mg/kg, and administrated intravenously five times daily. RESULTS: The correlation between the xenografts and hollow fiber assays was evaluated in 20 tumor cell lines and 4 anti-cancer agents. In the 20 tumor cell lines, the overall predictive accuracy of the hollow fiber assay for sensitivity was 83%, with a predictive accuracy for resistance of 92%. CONCLUSION: The hollow fiber assay was assessed as effective in drug efficacy evaluation, and found to be compatible with that of the xenografts assay.

Screening of brain-derived neurotrophic factor-like compounds / 中国药理学通报

Aim To apply simple combined models to screen small molecule BDNF-like compounds for potential pharmacotherapy of central nerve degenerative diseases.Methods BDNF-like bioactivities were dentified with effects against apoptosis induced by serum withdrawal in turn in cultured human SH-SY5y cells,rat NIH3T3/trkB cells vs NIH3T3 cells,using MTT technique.Results More than 400 compounds were tested by using these methods.11 of them presented protective effects on apoptosis induced by serum withdrawal in SH-SY5y cells.2 of them also provided selective protection against apoptosis induced in NIH3T3/trkB cells versus in NIH3T3 cells,which were identified as BDNF-like compounds.Further study showed that these 2 compounds could withstand SH-SY5y cell injuries induced by 6-OHDA.Conclusion All the above studies provide a useful series of simple models for BDNF-like drug screen and some compound candidates for further study of neuroprotection.