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

Tao Zhang; Yuanyuan Ding; Hongli An; Liuxin Feng; Sicen Wang

doi:10.1002/jssc.201500488

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

J Sep Sci. 2015 Sep;38(18):3247-3253. doi: 10.1002/jssc.201500488.

Authors

Tao Zhang¹, Yuanyuan Ding¹, Hongli An², Liuxin Feng³, Sicen Wang¹

Affiliations

¹ School of Medicine, Xi'an Jiaotong University, Xi'an, China.
² Center for Translational Medicine, The First Affiliated Hospital, Xian Jiaotong University College of Medicine, Xi'an, China.
³ Medical Laboratory, Second Affiliated Hospital, School of Medicine, Xi'an Jiaotong University, Xi'an, China.

PMID: 26178081
DOI: 10.1002/jssc.201500488

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.

Keywords: Cell membrane chromatography; Fibroblast growth factor receptor 4; High-performance liquid chromatography with mass spectrometry; Levistolide A; Mutations.