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目的 观察血管内皮生长因子 (VEGF)刺激上皮性卵巢癌细胞株Caov 3(Caov 3细胞 )后 ,信号转导及转录激活因子 3(STAT3)磷酸化的变化 ,探讨磷酸化的STAT3是否参与Caov 3细胞VEGF的信号转导。方法 采用免疫细胞化学和蛋白印迹的方法 ,测定体外不同浓度VEGF刺激Caov 3细胞STAT3磷酸化的水平 ;并观察能与VEGF受体 2 (VEGFR2 )特异性结合的短肽 ,对VEGF刺激STAT3磷酸化的阻断作用。结果 VEGF能够刺激Caov 3细胞的STAT3磷酸化。采用免疫细胞化学和蛋白印迹方法 ,浓度为 5 0ng/ml的VEGF刺激Caov 3细胞 30min后 ,Caov 3细胞STAT3磷酸化水平均增高 ,分别为 2 2 0± 0 2 8和 1 37± 0 17,与VEGF浓度为 0ng/ml时Caov 3细胞STAT3磷酸化的水平 (分别为 0 5 6± 0 15和 0 4 7± 0 19)比较 ,差异有极显著性 (P <0 0 0 1) ;并且STAT3磷酸化蛋白发生向细胞核内移位。VEGF刺激Caov 3细胞 6 0min后 ,Caov 3细胞STAT3磷酸化的水平 (分别为 0 95± 0 18和 0 6 6± 0 2 0 ) ,与刺激 30min时Caov 3细胞STAT3磷酸化的水平 (分别为 2 0 3± 0 17和 1 5 8±0 2 3)比较 ,差异有极显著性 (P <0 0 0 1)。与VEGFR2特异性结合的 80 μmol/L的短肽能有效抑制VEGF诱导STAT3的磷酸化水平。结论 VEGF在体外可诱导Caov 3?
Objective To observe the changes of phosphorylation of signal transducers and activators of transcription 3 (STAT3) after vascular endothelial growth factor (VEGF) stimulates Caov 3 cells and explore whether phosphorylated STAT3 is involved in Caov 3 Cell VEGF signaling. Methods Immunocytochemistry and Western blotting were used to determine the level of STAT3 phosphorylation in Caov 3 cells stimulated by different concentrations of VEGF in vitro. Short peptides with specific binding to VEGF receptor 2 (VEGFR2) were observed and the effects of VEGF on STAT3 phosphorylation Blocking effect. Results VEGF could stimulate STAT3 phosphorylation in Caov 3 cells. The level of STAT3 phosphorylation in Caov 3 cells after stimulated by VEGF at a concentration of 50ng / ml for 30min by immunocytochemistry and Western blotting was increased to 220 ± 0 2 8 and 1 37 ± 0 17, respectively Compared with the level of STAT3 phosphorylation in Caov 3 cells (0 56 ± 0 15 and 0 47 ± 0 19, respectively) at 0 ng / ml VEGF level, the difference was significant (P 0 01); and STAT3 phosphorylation occurs to the nucleus. The level of STAT3 phosphorylation in Caov 3 cells after treated with VEGF for 60 min was 0 95 ± 0 18 and 0 6 6 ± 0 0 0, respectively. The level of STAT3 phosphorylation in Caov 3 cells was 2 0 3 ± 0 17 and 1 58 ± 0 2 3), the difference was significant (P <0 0 0 1). The 80 μmol / L short peptides that specifically bind to VEGFR2 effectively inhibited the phosphorylation of STAT3 induced by VEGF. Conclusion VEGF can induce Caov 3 in vitro.