Construction of Y376C-FGFR4 eukaryotic expression plasmid and its biological activity in HEK293 cell

Jing, Huang; Feng, Liuxin; An, Hongli; Zhang, Tao

doi:10.1093/abbs/gmt091

Cited by 4 publications

(2 citation statements)

References 19 publications

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“…Based on the close relationship between FGFR4 receptors and cancer, Bu et al. [159] used a previously engineered HEK293 cell transfected with the Y367C mutation [160] to produce a mutant FGFR4‐expressing CM‐coated silica particle to screen a TCM product. It is important to note that the FGFR4 Y367C mutation is a dominant oncogene in MDA‐MB453 breast cancer cells [161] and as such, it is a good model for the screening of anticancer compounds through CM ligand fishing assay.…”

Section: Resultsmentioning

confidence: 99%

Ligand fishing as a tool to screen natural products with anticancer potential

Filho

Oliveira

Leal

et al. 2023

J of Separation Science

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Cancer is the second leading cause of death in the world and its incidence is expected to increase with the aging of the world's population and globalization of risk factors. Natural products and their derivatives have provided a significant number of approved anticancer drugs and the development of robust and selective screening assays for the identification of lead anticancer natural products are essential in the challenge of developing personalized targeted therapies tailored to the genetic and molecular characteristics of tumors. To this end, a ligand fishing assay is a remarkable tool to rapidly and rigorously screen complex matrices, such as plant extracts, for the isolation and identification of specific ligands that bind to relevant pharmacological targets. In this paper, we review the application of ligand fishing with cancer‐related targets to screen natural product extracts for the isolation and identification of selective ligands. We provide critical analysis of the system configurations, targets, and key phytochemical classes related to the field of anticancer research. Based on the data collected, ligand fishing emerges as a robust and powerful screening system for the rapid discovery of new anticancer drugs from natural resources. It is currently an underexplored strategy according to its considerable potential.

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Section: Resultsmentioning

confidence: 99%

Ligand fishing as a tool to screen natural products with anticancer potential

Filho

Oliveira

Leal

et al. 2023

J of Separation Science

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“…FGFR4 cells were constructed previously in our laboratory 43 . The cell culture medium was made up of DMEM supplemented with 10% fetal bovine serum and 100 μg mL −1 of streptomycin.…”

Section: Methodsmentioning

confidence: 99%

A novel cell membrane affinity sample pretreatment technique for recognition and preconcentration of active components from traditional Chinese medicine

et al. 2017

Sci Rep

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We describe a novel biomembrane affinity sample pretreatment technique to quickly screen and preconcentrate active components from traditional Chinese medicine (TCM), which adopts cell membrane coated silica particles (CMCSPs) as affinity ligands which benefit the biomembrane’s ability to maximize simulation of drug-receptor interactions in vivo. In this study, the prepared CMCSPs formed by irreversible adsorption of fibroblast growth factor receptor 4 (FGFR4) cell membrane on the surface of silica were characterized using different spectroscopic and imaging instruments. Drug binding experiments showed the excellent adsorption rate and adsorption capacity of FGFR4/CMCSPs compared with non-coated silica particles. The FGFR4/CMCSPs were used as solid-phase extraction sorbents to pretreat the TCM Aconitum szechenyianum Gay. The resultant FGFR4/CMCSPs exhibited good performance. In addition, high selectivity and recognition ability of the FGFR4/CMCSPs were determined by selectivity experiments. Four alkaloid were screened and identified, one of these alkaloid, napellonine, showed favorable anti-tumor activity in preliminary pharmacological verification trials including cell proliferation and molecular docking assays. The proposed cell membrane affinity sample pretreatment method is a reliable, effective and time-saving method for fast screening and enriching active compounds and can be extended to pretreat other TCMs as leading compounds resources.

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Screening anti‐tumor compounds from Ligusticum wallichii using cell membrane chromatography combined with high‐performance liquid chromatography and mass spectrometry

Zhang

Ding

et al. 2015

J of Separation Science

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Tyrosine 367 Cysteine-fibroblast growth factor receptor 4 cell membrane chromatography combined with high-performance liquid chromatography and mass spectrometry was developed. Tyrosine 367 Cysteine-HEK293 cells were used as the cell membrane stationary phase. The specificity and reproducibility of the cell membrane chromatography was evaluated using 1-tert-butyl-3-{2-[4-(diethylamino)butylamino]-6-(3,5-dimethoxyphenyl)pyrido[2,3-d]pyrimidin-7-yl}urea, nimodipine and dexamethasone acetate. Then, anti-tumor components acting on Tyrosine 367 Cysteine-fibroblast growth factor receptor 4 were screened and identified from extracts of Ligusticum wallichii. Components from the extract were retained on the cell membrane chromatographic column. The retained fraction was directly eluted into high-performance liquid chromatography with mass spectrometry system for separation and identification. Finally, Levistolide A was identified as an active component from Ligusticum wallichii extracts. The 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-tetrazolium bromide-formazan colorimetric assay revealed that Levistolide A inhibits proliferation of overexpressing the mutated receptor cells with dose-dependent manner. Phosphorylation of fibroblast growth factor receptor 4 was also decrease under Levistolide A treatment. Flex dock simulation verified that Levistolide A could bind with the tyrosine kinase domain of fibroblast growth factor receptor 4. Therefore, Levistolide A screened by the cell membrane chromatography combined with high-performance liquid chromatography and mass spectrometry can arrest cell growth. In conclusion, the two-dimensional high-performance liquid chromatography method can screen and identify potential anti-tumor ingredients that specifically act on the tyrosine kinase domain of the mutated fibroblast growth factor receptor 4.

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Construction of Y376C-FGFR4 eukaryotic expression plasmid and its biological activity in HEK293 cell

Cited by 4 publications

References 19 publications

Ligand fishing as a tool to screen natural products with anticancer potential

Ligand fishing as a tool to screen natural products with anticancer potential

A novel cell membrane affinity sample pretreatment technique for recognition and preconcentration of active components from traditional Chinese medicine

Screening anti‐tumor compounds from Ligusticum wallichii using cell membrane chromatography combined with high‐performance liquid chromatography and mass spectrometry

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