ABSTRACT
The presence of annexin II (Ann-II) during the initial stages of bovine embryo development and the regulation of Ann-II expression by retinol and insulin-like growth factor I (IGF-I) were studied. Bovine embryos at different stages of development were produced in vitro on Synthetic Oviductal Fluid (SOF) medium (control group), SOF supplemented with retinol (retinol group; 0.1ng/ml), or IGF-I (IGF-I group; 10ng/ml). The embryos were processed for mRNA extraction, cDNA production and polymerase chain reaction (PCR) using Ann-II-specific oligonucleotides. Ann-II was detected in all stages of early embryo development, except for the 16-cell stage. The blastocyst rates were significantly higher (P<0.05) in the group supplemented with retinol (37.8 percent, 45/119) during in vitro embryo culture (IVC) than in those cultured in SOF (20.5 percent, 24/117) or SOF with IGF-I (25.8 percent, 24/93). Semiquantitative analysis of Ann-II expression in embryos produced in medium supplemented with IGF-I or retinol revealed a lower expression of this gene when compared with embryos cultured in SOF (P<0.05). The Ann-II expression was not different in embryos cultured in the presence of retinol and IGF-I. The presence of retinol increased the production of embryos in vitro by decreasing the expression of Ann-II in early-stage of bovine embryo
Foram estudadas a presença de anexina II (Ann-II) durante a fase inicial do desenvolvimento embrionário bovino e sua regulação pelo retinol e pelo fator de crescimento semelhante à insulina (IGF-I). Embriões bovinos em diferentes estádios de desenvolvimento foram produzidos in vitro em fluido sintético de oviduto (SOF) sem suplementação (grupo-controle) ou suplementado com retinol (grupo retinol; 0,1ng/ml medium) ou IGF-I (grupo IGF-I; 10 ng/ml de meio). Os embriões foram processados para extração de mRNA, produção de cDNA e posterior análise por reação em cadeia da polimerase (PCR) com oligonucleotídeos específicos para Ann-II. Em todos os estádios de desenvolvimento embrionário, Ann-II foi detectada, exceto no estádio de 16 células. Os índices de blastocisto foram significativamente maiores (P<0,05) no grupo suplementado com retinol (37,8 por cento, 45/119) durante o cultivo in vitro de embriões (PIV) que aqueles obtidos no grupo controle (20,5 por cento, 24/117) ou no IGF-I (25,8 por cento, 24/93). Análise semiquantitativa da expressão de Ann-II em embriões produzidos em meio suplementado com IGF-I ou retinol revelou uma menor expressão desse gene quando comparado com embriões cultivados somente em SOF (P<0,05). A expressão de Ann-II não foi diferente em embriões cultivados na presença de retinol e IGF-I. A presença de retinol aumentou a produção de embriões in vitro, e diminuiu a expressão de Ann-II em estádios iniciais do desenvolvimento embrionário bovino
Subject(s)
Animals , Pregnancy , Cattle , /blood , Embryonic Development , Vitamin A/therapeutic useABSTRACT
OBJECTIVE: The aim of this study was to determine annexin II expression in cervical cancer. METHODS: In Ewha Womans University Mokdong Hospital, normal and cervical cancer tissues were obtained from healthy women (n=11), from patients with cervical intraepithelial neoplasia (CIN, n=11) and from patients with cervical cancer (n=33). The expressions of annexin II mRNA and protein were examined by quantitative competitive-PCR and by western blot analysis. Annexin II mRNA and protein expressions were examined with repects to the clinical characteristics including tumor sizes and cancer stages. RESULTS: The expression of annexin II mRNA in cervical cancer was higher than that in the normal cervix, and CIN (p0.05). The expression of annexin II protein in cervical cancer was higher than that in CIN but its expression was decreased according to the cervical cancer stage. CONCLUSION: Our results suggest that overexpression of annexin II mRNA and protein may be a biologic marker of cervical carcinogenesis.
Subject(s)
Female , Humans , Annexin A2 , Biomarkers , Blotting, Western , Carcinogenesis , Uterine Cervical Dysplasia , Cervix Uteri , RNA, Messenger , Uterine Cervical NeoplasmsABSTRACT
EGF has been a representive growth factor for understanding the signal transduction that underlies the biological response of a number of growth factors. EGF plays an important role in the regulation of growth inhibition in the squamous cell carcinoma with over expressed EGF receptors, but the mechanism that determine sensitivity to the growth inhibition by EGF are not well understood. The KUMA3 parental cell line, derived from a squamous cell carcinoma of the human lower lip, was established and variant cell line derived from the parental cell line was acquired by treatment with high dose EGF (200 ng/mL) for 6 months. The KUMA3 parental cells treated with EGF showed marked scattering cells with prominent dendritic processes. On the other hand, EGF treated cells in the variant cell line showed a dense monolayer of ovoid cells similar to untreated cells. Therefore, it was conformed that the variant cell line was resistant to the growth inhibitory effect of EGF. To gain insight into the action mechanism of EGF in the KUMA3 parental and variant cell lines, Western blot analysis was examined to identify transregulation of EGF receptor. In the all parental cell lines treated with EGF, EGF receptor proteins were completely disappeared by lysosomal degradation. However, EGF receptor proteins were shown the periodic appearance according to the time course in the variant cell lines treated with EGF. In Western blot and immunocytochemical analyses, annexin II protein evenly distributed in the cytoplasm relocated to the plasma membrane at 9 hr exposure to EGF and was significantly decreased at 12 hr and 24 hr exposures to EGF in the variant cell line. However, annexin II protein relocated to the nuclear membrane at 9 hr exposure to EGF and was not any change in amount at 12 hr and 24 exposure to EGF in the parental cell line. In conclusion, this study suggests that the retention of colony formation pattern and cell morphology in the variant cell line treated with EGF might be owing to the heterologous desensitization of EGF receptor and exocytosis of annexin II.