Journal
JOURNAL OF CHROMATOGRAPHY B-ANALYTICAL TECHNOLOGIES IN THE BIOMEDICAL AND LIFE SCIENCES
Volume 912, Issue -, Pages 85-92Publisher
ELSEVIER SCIENCE BV
DOI: 10.1016/j.jchromb.2012.10.029
Keywords
Fibroblast growth factor receptor 4; Cell membrane chromatography; High performance liquid chromatography/mass spectrometry; Brassica albla L.; Sinapine
Funding
- National Natural Science Foundation of China [81202495]
- Program for New Century Excellent Talents in University [NCET-08-0437]
- Fundamental Research Funds for the Central University
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We investigated an analytical method for the recognition separation, and identification of active components from the traditional Chinese medicinal plant Brassica albla L. using fibroblast growth factor receptor 4 cell membrane chromatography (FGFR4/CMC) with high performance liquid chromatography/mass spectrometry (HPLC/MS). HEK293-FGFR4 cells were obtained by stable transfection of the HEK293 cell line with pcDNA3.1 vector containing the FGFR4 gene. Crude extracts of B. albla L. were firstly subjected to FGFR4/CMC column, and the retain contents on the FGFR4/CMC column were then transferred and enriched using a pre-column, and a ten port column switcher were used between FGFR4/CMC column and HPLC. The retained components on FGFR4/CMC column were then directly delivered to the HPLC/MS system for separation and identification. Gefitinib, nicotine, atenolol, and nimodipine were used in order to verify FGFR4/CMC-HPLC/MS system specificity. Subsequently, we investigated the reproducibility and reliability of the FGFR4/CMC-HPLC/MS system. Finally, sinapine was identified as an active component of B. albla L The MTT calorimetric assay revealed sinapine could inhibit the proliferation of HEK293-FGFR4 cells with dose dependence. Competitive displacement assay approved getitinib could occupy binding site of sinapine with competition way. And FleX dock simulation further exhibited sinapine and gefitinib could bind with the FGFR4 tyrosine active domain. Thus, sinapine is a potential tumor antagonist that acts on the tyrosine kinase domain. (C) 2012 Elsevier B.V. All rights reserved.
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