title: Screening anti-tumor compounds from Ligusticum wallichii using cell membrane chromatography combined with high-performance liquid chromatography and mass spectrometry.
abstract: 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.
authors.affiliation: School of Medicine, Xi'an Jiaotong University, Xi'an, China.
authors.forename: Tao
authors.initials: T
authors.lastname: Zhang
authors.affiliation: School of Medicine, Xi'an Jiaotong University, Xi'an, China.
authors.forename: Yuanyuan
authors.initials: Y
authors.lastname: Ding
authors.affiliation: Center for Translational Medicine, The First Affiliated Hospital, Xian Jiaotong University College of Medicine, Xi'an, China.
authors.forename: Hongli
authors.initials: H
authors.lastname: An
authors.affiliation: Medical Laboratory, Second Affiliated Hospital, School of Medicine, Xi'an Jiaotong University, Xi'an, China.
authors.forename: Liuxin
authors.initials: L
authors.lastname: Feng
authors.affiliation: School of Medicine, Xi'an Jiaotong University, Xi'an, China.
authors.forename: Sicen
authors.initials: S
authors.lastname: Wang
keywords: Cell membrane chromatography; Fibroblast growth factor receptor 4; High-performance liquid chromatography with mass spectrometry; Levistolide A; Mutations