雌激素及其阻滞剂对Ishikawa细胞中ETS-1和VEGF调控的研究

目的:探讨雌激素及其阻滞剂对子宫内膜癌细胞系Ishikawa细胞中转录因子ETS-1和血管内皮细胞生长因子(VEGF)表达的调控机制。方法:用RT-PCR法检测分别使用不同浓度雌激素、雌激素受体阻滞剂氟维司群处理后Ishikawa细胞中ETS-1基因表达水平的变化;用细胞爬片免疫组化法检测分别使用这两种不同浓度药物后Ishikawa细胞中ETS-1与VEGF表达的差别;采用ELISA法检测分别使用两种不同浓度的药物后Ishikawa细胞培养液中VEGF蛋白表达量的变化。结果:(1)随着雌激素浓度增加,ETS-1基因表达上调(FP=3.84,P<0.05);随着氟维司群浓度增加,ETS-1基因表达下调(FP=7.01,P<0.01);(2)随着雌激素浓度增加,ETS-1(FP=3.72,P<0.05)和VEGF(FP=3.68,P<0.05)蛋白质水平表达都增加;随着氟维司群浓度增加,ETS-1(FP=4.37,P<0.05)和VEGF(FP=4.09,P<0.05)蛋白质水平的表达都减少;(3)随着雌激素浓度增加,分泌型VEGF蛋白表达量上升(FP=3.91,P<0.05);随着氟维司群浓度增加,分泌型VEGF蛋白表达量下调(FP=7.34,P<0.01)。结论:雌激素可促进Ishikawa细胞中ETS-1和VEGF的表达,VEGF表达与ETS-1表达呈正相关。

The study of regulation of ETS-1 and VEGF by estrogen and estrogen receptor blocker in endometrial carcinoma cell line Ishikawa

Objective:To explore the role of E2 and it's receptor blocker in the regulations of ETS-1 and VEGF genes expression in ER-positive human endometrial carcinoma cell line Ishikawa.Methods:ETS-1 gene expression in Ishikawa cells was detected by RT-PCR after the treatments of either estrogen or fulvestrant in different concentrations.The expression differences of ETS-1 and VEGF in Ishikawa cells were detected by immunohistochemical technique following the same treatment.Moreover,the secretion of VEGF protein in the culture media of Ishikawa cells was also detected by ELISA.Result:The expressions of ETS-1 were up regulated by estrogen(FP=3.84,P<0.05),reversely,the expressions of ETS-1 were down regulated with the increasing concentrations of fulvestrant(FP=7.01,P<0.01).The expressions of ETS-1(FP=3.72,P<0.05) and VEGF(FP=3.68,P<0.05) at protein level were increased by estrogen,while,the expressions of ETS-1(FP=4.37,P<0.05) and VEGF(FP=4.09,P<0.05) at protein level were decreased with the addition of fulvestrant.Furthermore,the secretions of VEGF protein in culture media were increased with estrogen(FP=3.91,P<0.05),in contrast,VEGF protein concentrations were declined with the supplement of fulvestrant in the culture media(FP=7.34,P<0.01).Conclusion:The expressions of ETS-1 and VEGF can be regulated by estrogen and the expression of VEGF is related to ETS-1 in endometrial carcinomal cell line Ishikawa.