Overexpression of FOXG1 contributes to TGF-β resistance through inhibition of p21WAF1/CIP1 expression in ovarian cancer

Background:Loss of growth inhibitory response to transforming growth factor-Β (TGF-Β) is a common feature of epithelial cancers. Recent studies have reported that genetic lesions and overexpression of oncoproteins in TGF-Β/Smads signalling cascade contribute to the TGF-Β resistance. Here, we showed that the overexpressed FOXG1 was involved in attenuating the anti-proliferative control of TGF-Β/Smads signalling in ovarian cancer.Methods:FOXG1 and p21 WAF1/CIP1 expressions were evaluated by real-time quantitative reverse-transcription polymerase chain reaction (RT-PCR), western blot and immunohistochemical analyses. The effect of FOXG1 on p21 WAF1/CIP1 transcriptional activity was examined by luciferase reporter assays. Cell lines stably expressing or short hairpin RNA interference-mediated knockdown FOXG1 were established for studying the gain-or-loss functional effects of FOXG1. XTT cell proliferation assay was used to measure cell growth of ovarian cancer cells.Results:Quantitative RT-PCR and western blot analyses showed that FOXG1 was upregulated and inversely associated with the expression levels of p21 WAF1/CIP1 in ovarian cancer. The overexpression of FOXG1 was significantly correlated with high-grade ovarian cancer (P0.025). Immunohistochemical analysis on ovarian cancer tissue array was further evidenced that FOXG1 was highly expressed and significantly correlated with high-grade ovarian cancer (P0.048). Functionally, enforced expression of FOXG1 selectively blocked the TGF-Β-induced p21 WAF1/CIP1 expressions and increased cell proliferation in ovarian cancer cells. Conversely, FOXG1 knockdown resulted in a 20-26% decrease in cell proliferation together with 16-33% increase in p21 WAF1/CIP1 expression. Notably, FOXG1 was able to inhibit the p21 WAF1/CIP1 promoter activity in a p53-independent manner by transient reporter assays.ConclusionOur results suggest that FOXG1 acts as an oncoprotein inhibiting TGF-Β-mediated anti-proliferative responses in ovarian cancer cells through suppressing p21 WAF1/CIP1 transcription. © 2009 Cancer Research UK All rights reserved.published_or_final_versio

Purification and molecular structure of two digalactosyl D-chiro-inositols and two trigalactosyl D-chiro-inositols from buckwheat seeds

Two digalactosyl D-chiro-inositols and two trigalactosyl D-chiuo-inositols, members of the fagopyritol A series and fagopyritol B series, were isolated from buckwheat (Fagopyrum esculentum Moench) seeds. Structures of the first three were determined by H-1 and C-13 NMR. Fagopyritol B2 is alpha -D-galactopyranosyl-(1->6)-alpha -D-galactopyranosyl-(1->2)-1D-chiro-inositol , and fagopyritol A2 is alpha -D-galactopyranosyl-(1->6)-alpha -D-galactopyranosyl-(1->3)-1D-chiro-inositol. Fagopyritol A3, a trigalactosyl D-chiro-inositol, is alpha -D-galactopyranosyl-(1->6)-alpha -D-galacto-pyranosyl-(1->6)-alpha -D-galactopyranosyl-(1->3)-1D-chiro-inositol. From analysis of hydrolysis products, the second trigalactosyl D-chiro-inositol, fagopyritol B3, is alpha -D-galactopyranosyl-(1->6)-alpha -D-galactopyranosyl-(1->6)-alpha -D-galactopyranosyl-(1->2)-1D-chiro-inositol. (C) 2001 Elsevier Science Ltd. All rights reserved

Fagopyritols, D-chiro-inositol, and other soluble carbohydrates in buckwheat seed milling fractions

Fagopyritols are mono-, di-, and trigalactosyl derivatives of D-chiro-inositol that accumulate in seeds of common buckwheat (Fagopyrum esculentum Moench) and may be important for seed maturation and as a dietary supplement. Fagopyritols and other soluble carbohydrates were assayed in mature greats and 11 milling fractions of common buckwheat seed. Because fagopyritols are in embryo and aleurone tissues, differences in fagopyritol concentrations reflect varying proportions of these tissues in each milling fraction. Bran milling fractions contained 6.4 g of total soluble carbohydrates per 100 g of dry weight, 55% of which was sucrose and 40% fagopyritols. Flour milling fractions had reduced fagopyritol concentration [0.7 g/100 g of dry weight total fagopyritols in the dark (Supreme) flour and 0.3 g/100 g in the light (Fancy) flours]. Fagopyritol B1 was 70% of total fagopyritols in all milling fractions. Fagopyritols were 40% of total soluble carbohydrates in greats of two cultivars of common buckwheat but 21% in groats of tartary buckwheat [Fagopyrum tataricum (L.) Gaertn.], probably a reflection of environment and genetics. A rhamnoglucoside present in tartary buckwheat was not detected in common buckwheat

Molecular structure of fagopyritol Al (O-alpha-D-galactopyranosyl-(1 -> 3)-D-chiro-inositol) by NMR

The molecular structure of fagopyritol A1, a novel galactopyranosyl cyclitol from buckwheat seeds, was determined to be O-alpha-D-galactopyranosyl-(1-->2)-D-chiro-inositol by H-1 and C-13 NMR. Fagopyritol Al is a positional isomer of fagopyritol B1 (O-alpha-D-galactopyranosyl-(1-->2)-D-chiro-inositol), representing a different series of fagopyritol oligomers. Trimethylsilyl derivatives of both compounds have similar mass spectra, but each may be identified by different abundance ratios of fragments with m/z 305/318 and 318/319. (C) 2000 Elsevier Science Ltd. All rights reserved