RESUMEN
Objective To observe the effect of the total flavonoids of Clerodendrum Bungei (TFCB) on proliferation, invasion and migration of A549 cell lines by Wnt/β-catenin signal pathway. Methods Transfecting A549 cell lines with empty vector pcDNA3.1 and β-catenin plasmid which has the mutation at S45 (S45A-β-catenin), which were intervened with TFCB. MTT assay detected cell proliferation, scratch test observed cell migration, Transwell experiment detected the cell invasion. The expression of β-catenin, E-Cadherin, vimentin, Slug, GSK-3β, and p-GSK-3β protein in each group was detected by Western blotting. Results The proliferation, invasion and migration of A549 cells were enhanced significantly after transfected with S45A-β-catenin plasmid (P < 0.05 or 0.01), along with the increasing expression of β-catenin, vimentin, and the reducing expression of E-cadherin, GSK-3β, P-GSK-3β (P < 0.01). TFCB can inhibit the proliferation, migration and invasion of A549 cells (P < 0.05), expecially in S45A-β-catenin group (P < 0.001). A549 cells transfected with empty vector had the ability of up-regulating the expression of E-cadherin, GSK-3β and P-GSK-3β, down-regulating the expression of β-catenin and vimentin with TFCB. A549 cells transfected with S45A-β-catenin plasmid had the ability of down-regulating the expression of β-catenin and vimentin with TFCB. Conclusion The mechanism of inhibiting lung cancer of TFCB maybe associate with the inhibitory expression of β-catenin and regulate the downstream factors, with view to inducing EMT by activating Wnt/β-catenin pathway.
RESUMEN
Ten compounds were isolated from the 95% aqueous EtOH extract of Clerodendrum bungei by a combination of various chromatographic techniques including column chromatography over silica gel, Sephadex LH-20, MCI, ODS, and semi-preparative HPLC. Their structures were elucidated as 11,12,16S-trihydroxy-7-oxo-17(15→16),18(4→3)-diabeo-abieta-3,8,11,13-tetraen-18-oic acid (1), 12S*,13R*-dihydroxy-9-oxo-octadeca-10(E)-enoic acid (2), clerodenoside A (3), trichotomoside (4), glycosmisic acid (5), 4'-O-methylscutellarein (6), neroplomacrol (7), butylitaconic acid (8), hexylitaconic acid (9), p-hydroxybenzonic acid (10) by their physicochemical properties and spectroscopic data. Compounds 1 and 2 are new natural products, while compounds 7-10 were obtained from the genus Clerodendrum for the first time, and compounds 3, 5, 6 were isolated from this plant for the first time.