Expression of Heat Shock Proteins and Cytokines in Response to Ethanol Induced Damage in the Small Intestine of ICR Mice

BACKGROUND/AIMS: Ethanol administration causes intestinal epithelial cell damage by increasing intestinal permeability and the translocation of endotoxins from intestinal bacterial flora. Heat shock proteins (HSPs) are associated with recovery and protection from cell damage. The aim of the current study was to investigate differences in the expression of HSPs in the small intestine and the biochemical changes attributable to ethanol-induced intestinal damage. METHODS: Ethanol (20%) was injected intraperitoneally (2.75 g/kg, 5.5 g/kg, 8.25 g/kg) in ICR mice and the same volume of saline was administered to controls. After 1 hour, the proximal, middle, and distal segments were taken from the small intestine and the degree of damage was analyzed. In each segment, the expression of HSPs was analyzed by western blotting. The expression of inflammatory mediators including interleukin-1beta (IL-1beta), tumor necrosis factor-alpha (TNF-alpha), cyclooxygenase-2 (COX-2), and antioxidant enzyme such as glutathione-S-transferase were compared using real-time polymerase chain reaction assays. RESULTS: In the control group, HSP70 increased in all segments of small intestine. Additionally, increases in the expression of HSP40 and HSP90 in the distal regions and an increase in HSP32 in the middle regions were observed. After ethanol treatment, greater histological damage was observed in the distal small intestine and significant decreases in HSPs were observed generally. Increased expression of IL-1beta, TNF-alpha, and COX-2 was observed in small intestinal tissues exposed to ethanol-induced damage. However, there was no significant difference in the expression of an antioxidant enzyme. CONCLUSIONS: Significant differences in the expression of HSPs in different intestinal regions were observed. These differences may have been attributable to the distribution of intestinal bacteria.

Microarray analysis of gene expression in mice ovaries exposed to a 1.765 GHz microwave in utero / 대한산부인과학회지

OBJECTIVE: The aim of this study was to evaluate the effects on murine fetal ovarian gene expression of prenatal exposure to 1.765 GHz of microwave irradiation. METHODS:Ten pregnant ICR mice were divided into two groups. At 5th days after mating, dam mice were exposed to microwave (SAR: 0.38~1.71 W/kg) in the insulated cage for 8 hours each day. The remaining mice were treated in the same way. Neonatal ovaries were removed for study 7 days after delivery. Microarray analysis was performed using total RNA extracted from the removed ovaries. We investigated the differences in ovarian gene expression between the groups. SPSS 12.0 was used for statistical analysis. P<0.05 was considered to be statistically significant. RESULTS: The mean birth weight of the offspring in the irradiated group was significantly lower than that in the sham group (1.54+/-.22 g vs. 1.60+/-.21 g, P=0.012). The mean number of offspring per pregnancy in the irradiated group was significantly higher than in the sham group (13.60+/-.70 vs. 11.40+/-.17, P=0.009). We detected that in the irradiated ovaries, 14 genes were expressed at levels 2-fold higher than in the sham ovaries and 74 genes were expressed at levels 2-fold lower than in the sham ovaries. CONCLUSION: We found differences in fetal ovarian gene expression between the irradiated and sham groups. In the irradiated group, the Tnfaip8, TNFsf 12, Cfd, CCL 11, and Zfp74 genes were down-regulated and the Brd 3 gene was up-regulated.

Microarray analysis of gene expression in mice ovaries exposed to a 1.765 GHz microwave in utero / 대한산부인과학회지

OBJECTIVE: The aim of this study was to evaluate the effects on murine fetal ovarian gene expression of prenatal exposure to 1.765 GHz of microwave irradiation. METHODS:Ten pregnant ICR mice were divided into two groups. At 5th days after mating, dam mice were exposed to microwave (SAR: 0.38~1.71 W/kg) in the insulated cage for 8 hours each day. The remaining mice were treated in the same way. Neonatal ovaries were removed for study 7 days after delivery. Microarray analysis was performed using total RNA extracted from the removed ovaries. We investigated the differences in ovarian gene expression between the groups. SPSS 12.0 was used for statistical analysis. P<0.05 was considered to be statistically significant. RESULTS: The mean birth weight of the offspring in the irradiated group was significantly lower than that in the sham group (1.54+/-.22 g vs. 1.60+/-.21 g, P=0.012). The mean number of offspring per pregnancy in the irradiated group was significantly higher than in the sham group (13.60+/-.70 vs. 11.40+/-.17, P=0.009). We detected that in the irradiated ovaries, 14 genes were expressed at levels 2-fold higher than in the sham ovaries and 74 genes were expressed at levels 2-fold lower than in the sham ovaries. CONCLUSION: We found differences in fetal ovarian gene expression between the irradiated and sham groups. In the irradiated group, the Tnfaip8, TNFsf 12, Cfd, CCL 11, and Zfp74 genes were down-regulated and the Brd 3 gene was up-regulated.

Effects of the Specific COX-2 Inhibitor, Celecoxib, on Paclitaxel-Induced Apoptosis in SK-OV-3 Epithelial Ovarian Cancer Cell Line / 대한산부인과학회지

OBJECTIVE: In vitro studies have revealed that treatment of various human cancer cell lines with specific cyclooxygenase 2 (COX-2) inhibitors induces apoptotic cell death. The goal of this article is to investigate the benefits of combining COX-2 inhibitors with existing treatment modalities in the management of ovarian cancer. METHODS: In this study we sought to determine the effects of combining paclitaxel and the COX-2 inhibitor celecoxib on apoptosis of epithelial ovarian cancer (EOC) cells. SK-OV-3 cells were exposed to increasing concentrations of paclitaxel (10(-7) M, 10(-6) M and 10(-5) M) and celecoxib (10(-8) M, 10(-7) M, 10(-6) M, 10(-5) M and 10(-4) M) as well as a combination of both drugs. The activity of apoptosis was evaluated by the morphologic examination and the MTT assay. The pattern of apoptosis was also assessed by the caspase-3 activity and the fraction of cleaved PARP (poly ADP-ribose polymerase) protein. RESULTS: Single application of both drugs could significantly increase the rate of apoptosis after 24 hours of continuous exposure. But concomitant treatment of SK-OV-3 EOC cell line with paclitaxel and celecoxib resulted in marked impairment of paclitaxel-induced apoptosis. The pattern of apoptosis induced by paclitaxel on SK-OV-3 EOC cell line was caspase-3 independent. CONCLUSION: Combining COX-2 inhibitors and paclitaxel does not have an additive or synergistic tumoricidal effect. On the contrary, celecoxib treatment markedly inhibited the apoptotic effects of paclitaxel in SK-OV-3 EOC cell line.

In Vitro Responses of SK-OV-3 Ovarian Cancer Cell Lines to Tamoxifen and Celecoxib / 대한산부인과학회지

OBJECTIVE: There are some evidences that some epithelial ovarian cancer cells respond to hormonal therapy. And in vitro studies have revealed that treatment of various human cancer cell lines with selective cyclooxygenase 2 (COX-2) inhibitors induces apoptotic cell death. The goal of this article is to evaluate the effects of tamoxifen and celecoxib, a selective COX-2 inhibitor, on the ovarian cancer cells and the benefits of combining these agents in the management of ovarian cancer. METHODS: SK-OV-3 epithelial ovarian cancer cells were exposed to increasing concentration of tamoxifen (10(-8) M, 10(-7) M, 10(-6) M, 10(-5) M and 10(-4) M) and celecoxib (10(-8) M, 10(-7) M, 10(-6) M, 10(-5) M and 10(-4) M) as well as a combination of both drugs. The activity of apoptosis was evaluated by the morphologic examination and the MTT assay. The pattern of apoptosis was also assessed by the caspase-3 activity and the fraction of cleaved PARP (poly ADP-ribose polymerase) protein. RESULTS: Single application of both drugs could significantly increase the rate of apoptosis after 24 h of continuous exposure. Concomitant treatment of SK-OV-3 cells with tamoxifen and celecoxib induced significant increase in apoptosis, comparing with single drug exposure. The pattern of apoptosis induced by these agents on SK-OV-3 cells seemed to be caspase-3 dependent. CONCLUSION: Our data suggest that combining tamoxifen with selective COX-2 inhibitor seems to have at least an additive tumoricidal effect. A more definitive role for this combination therapy in clinical settings in ovarian cancer will need to be defined through the conduct of clinical trials.