5,7-Dihydroxy-4'-methoxyisoflavone induces apoptosis by inhibiting the ERK and Akt pathways in human osteosarcoma cells.

Phytoestrogens are known to prevent tumor progression by inhibiting proliferation and inducing apoptosis in cancer cells. In this study we determine the effect of 5,7-dihydroxy-4'-methoxyisoflavone, a phytoestrogen, on proliferation and apoptosis in the human osteosarcoma (OS) cell line U2OS. 5,7-Dihydroxy-4'-methoxyisoflavone dose-dependently inhibited proliferation in U2OS cells, which was accompanied by an increase of early apoptotic cells. However, 5,7-dihydroxy-4'-methoxyisoflavone had little effect on the growth and apoptosis of normal human skin fibroblast (HSF) cells. This may indicate that 5,7-dihydroxy-4'-methoxyisoflavone can selectively inhibit the proliferation of cancerous cells. Meanwhile, 5,7-dihydroxy-4'-methoxyisoflavone decreased the protein levels of phosphorylated ERK and Akt. Inactivation of these pathways was confirmed by upregulation of Bax expression and downregulation of Bcl-2 expression. Phosphorylated Akt protein levels were decreased in HSF cells only at a high concentration (80 µM) of 5,7-dihydroxy-4'-methoxyisoflavone. Together, we suggest that 5,7-dihydroxy-4'-methoxyisoflavone promotes cell death of human OS cells U2OS by induction of apoptosis, which is associated with the inhibition of ERK and Akt signaling. Thus, 5,7-dihydroxy-4'-methoxyisoflavone may have less toxicity compared to normal tissue and could be a potential therapy for OS.

[Impact of width of hepatectomy margin on survival after simultaneous liver and colorectal resection for colorectal cancer liver metastasis].

OBJECTIVE: To elucidate an adequate hepatectomy margin for simultaneous liver and colorectal resection in colorectal cancer liver metastasis. METHODS: Clinical data of 39 patients, undergone simultaneous liver and colorectal resection for colorectal cancer liver metastasis from August 1994 to December 2004, were analyzed retrospectively. Two groups were divided according to the width of hepatectomy margin:less than 1 cm in group A, and equal or more than 1 cm in group B. The data were analyzed and compared between the 2 groups using Kaplan-Meier survival analysis and Log-rank test. RESULTS: There were 14 patients in group A and 25 patients in group B. No significant differences in gender, age, primary tumor invasion, lymph node metastasis, the number, distribution and size of liver metastasis, duration and blood lose of surgery were found between two groups. The median survival time was 17 months in group A, and 37 months in group B, and the overall 5-year survival rate in group B was much better than that in group A (19.8% vs 0, P<0.01). CONCLUSION: Simultaneous liver and colorectal resection in colorectal cancer liver metastasis should be performed with a hepatectomy margin equal or more than 1 cm.

Effect of biofilm formation on virulence factor secretion via the general secretory pathway in Streptococcus mutans.

OBJECTIVES: To investigate the role of SecA in protein secretion, and to evaluate the effect of biofilm formation on protein secretion in Streptococcus mutans. DESIGN: S. mutans strains UA159 and GS-5 were used in this study. Cells grown in biofilm and planktonic conditions were observed using immunogold electron microscopy. The mRNA levels of ftf, gtfB, gtfC, gtfD, Pac and secA were analysed in different growth conditions using real-time quantitative polymerase chain reaction. The levels of wall proteins and whole-cell protein extracts were examined using Western blot analysis. RESULTS: A microdomain colocalised with SecA and virulence factors such as Pac (AgI/II) and glucosyltransferase (GTF) was observed. The mRNA level of secA was upregulated in the biofilm condition. The level of protein expression of SecA and wall protein levels of GTF, fructosyltransferase (FTF) and Pac (AgI/II) in the biofilm condition were significantly higher than in the planktonic condition. CONCLUSIONS: These data suggest that S. mutans utilises the Sec pathway to secrete virulence factor proteins such as Pac (AgI/II), GTF and FTF, and protein secretion occurred at a distinct microdomain. The level of SecA, the key factor in the Sec pathway, was influenced significantly by biofilm formation in S. mutans.

Sec translocase and sortase A are colocalised in a locus in the cytoplasmic membrane of Streptococcus mutans.

INTRODUCTION: The general secretory (Sec) pathway is the main pathway for protein transport. Sortase A has been shown to anchor surface protein containing an LPXTG motif to the cell wall. In order to know the distribution pattern of the Sec translocons and sortase A in Streptococcus mutans, we explored the subcellular localisation of SecA and sortase A. METHODS: The anti-SecA antibody and anti-sortase A antibody were examined on Western blots to evaluate specificities. An indirect post-embedding immunogold method was used to determine the subcellular localisation of the Sec pathway and sortase A enzyme in S. mutans membrane. RESULTS: Immunoblotting experiments showed that the anti-SecA antibody and anti-sortase A antibody specifically recognised a single band of about 95 kDa and 27 kDa, respectively in the S. mutans GS-5 cell lysates. Immunogold electron micrographs showed that small and large gold particles clustered together at a distinct microdomain. CONCLUSIONS: These data showed for the first time that Sec translocase and sortase A colocalised to a single locus in the cytoplasmic membrane of S. mutans GS-5.

Increased expression of collagen prolyl 4-hydroxylases in Chinese patients with hereditary gingival fibromatosis.

OBJECTIVES: Hereditary gingival fibromatosis (HGF) is characterized by excess accumulation of interstitial collagen. However, until now, there has been controversy about the mechanism of collagen accumulation in HGF gingivae. The present study aimed to clarify the pathogenic mechanisms potentially involved. DESIGN: Gingival fibroblasts from three Chinese HGF patients and three healthy subjects were cultured. Cell proliferation was assessed by MTT assay. The mRNA levels of type I collagen, MMP-1, MMP-3, TIMP-1, prolyl 4-hydroxylase (P4H)alpha(I), alpha(II), alpha(III) and P4Hbeta were analyzed in gingival fibroblasts by RT-PCR. The protein production of type I collagen and P4H was examined respectively by ELISA and Western blot. RESULTS: In culture, HGF gingival fibroblasts showed similar growth characteristics to fibroblasts isolated from control gingivae. The mRNA and protein levels of type I collagen and P4Halpha in HGF fibroblasts were higher than those in controls. There were no detected differences in mRNA expression levels of MMP-1, MMP-3, TIMP-1, P4Halpha(II), alpha(III) and P4Hbeta between HGF and control fibroblasts. CONCLUSIONS: These data suggest that increased collagen post-translational modification by P4H may be one mechanism by which increased collagen accumulation occurs in some forms of HGF.