Purpose: In this study, we examined the promoter methylation status and expression of 14-3-3 and evaluated its clinical significance in epithelial ovarian cancer.Experimental Design: Twelve ovarian cancer cell lines; 2 ovarian surface epithelial cell lines; and 8 normal, 8 benign, 12 borderline, and 102 ovarian cancer tissues were examined. Methylation-specific PCR, quantitative reverse transcription-PCR, and immunohistochemistry were used to evaluate methylation status and expression of 14-3-3 gene and protein.Results: Among the 12 ovarian cancer cell lines, the presence of a methylated band was detected in seven cell lines. Median values of relative 14-3-3 gene expression in cancers with methylation (3.27) were significantly lower than those without methylation (16.4; P < 0.001). Treatment of 5-aza-2-deoxycitidine resulted in the demethylation of the promoter CpG islands and reexpression. All of the normal, benign, and borderline tissues were positive for 14-3-3 protein, and in ovarian cancer tissues, 73.5% (75 of 102) were positive for 14-3-3 protein and was almost consistent with methylation status. Negative immunoreactivity of 14-3-3 was significantly correlated with high age and serous histology, high-grade, advanced-stage residual tumor of >2 cm, high serum CA125, high Ki-67 labeling index, and positive p53 immunoreactivity. 14-3-3 immunoreactivity was significantly associated with overall survival (P ‫؍‬ 0.0058).Conclusions: Our findings suggest that 14-3-3 is inactivated mainly by aberrant DNA methylation and that it may play an important role in the pathogenesis of epithelial ovarian cancer.

Heat shock (438C, 45 min) induced transient nuclear accumulation of p53 in primary human ®broblasts without any clonogenically toxic e ects. The accumulation of p53 reached a maximal level 3*5 h after heat shock, and returned to the basal level within 12 h. Following the increase in p53 level, cell cycle arrest at G1/S was observed in normal ®broblasts, whereas neither nuclear accumulation of p53 nor cell cycle arrest were observed in HeLa cells. By comparing cell cycle patterns of heat-treated mouse cells with di erent genotypes at the p53 locus (+/+, +/7, 7/7), the observed cell cycle arrest at G1/S was demonstrated to be p53-dependent. Cell cycle arrest in normal human ®broblasts continued for nearly 24 h, resulting in a one day delay of cell growth compared with non-treated cells. Following enhancement of the p53 level, the amount of p21/WAF1/ CIP1 increased, and the high level of p21 was sustained for almost one day in a cell cycle-independent manner, suggesting the involvement of p21 in the inhibition of cell cycle progression by heat shock.

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Aims:  To identify and characterize a new adhesin‐like protein of probiotics that show specific adhesion to human blood group A and B antigens. Methods and Results:  Using the BIACORE assay, the adhesion of cell surface components obtained from four lactobacilli strains that adhered to blood group A and B antigens was tested. Their components showed a significant adhesion to A and B antigens when compared to the bovine serum albumin (BSA) control. The 1 mol l−1 GHCl fraction extracted from Lactobacillus mucosae ME‐340 contained a 29‐kDa band (Lam29) using SDS–PAGE. The N‐terminal amino acid sequence and homology analysis showed that Lam29 was 90% similar to the substrate‐binding protein of the ATP‐binding cassette (ABC) transporter from Lactobacillus fermentum IFO 3956. The complete nucleotide sequence (858 bp) of Lam29 was determined and encoded a protein of 285 amino acid residues. Phylogenetic analysis and multiple sequence alignments indicated this protein may be related to the cysteine‐binding transporter. Conclusions:  The adhesion of ME‐340 strain to blood group A and B antigens was mediated by Lam29 that is a putative component of ABC transporter as an adhesin‐like protein. Significance and Impact of the Study:  Lactobacillus mucosae ME‐340 expressing Lam29 may be useful for competitive exclusion of pathogens via blood group antigen receptors in the human gastrointestinal mucosa and in the development of new probiotic foods.

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