摘要
目的:利用HER-2/neu转染前后的Ishikawa细胞株,探讨HER-2/neu对雌激素依赖性子宫内膜癌细胞信号通路(MAPK/ERK、PI3K/Akt)的调控作用。方法:表皮生长因子(endothelial growth factor)处理HER-2/neu转染前后的Ishikawa细胞株,Western-blot检测转染前后细胞COX-2、t-Akt及p-Akt蛋白、t-ERK及p-ERK表达,ELISA方法检测细胞培养上清液中雌二醇的含量。用PI3K/Akt的抑制剂LY294002及MAPK/ERK的抑制剂PD98059抑制信号通路,以不同浓度作用相同时间和以相同浓度分别作用不同时间后,检测COX-2及雌二醇表达水平。结果:转染HER-2/neu的Ishikawa细胞株COX-2蛋白、p-Akt蛋白、p-ERK及细胞上清液中的雌二醇的含量均高于未转染的Ishikawa细胞株(P<0.05)。应用抑制剂分别抑制上述两种通路,转染后细胞株中COX-2的表达水平低于转染前Ishikawa细胞株,同时转染组细胞上清液中雌二醇的含量较未转染组下降明显,降幅更大。随抑制剂浓度的增加及作用时间的延长,两组细胞COX-2及雌二醇的表达均逐渐减少,且抑制作用与浓度及作用时间呈负相关关系。结论:子宫内膜癌Ishikawa细胞株HER-2/neu基因表达的增强进一步引起COX-2、E2表达的增加,可能是通过对信号通路MAPK/ERK、PI3K/Akt的调控而实现的。
Objective ：To use HER -2/neu transfected Ishikawa cell line before and after, HER -2/neu estrogen - dependent endometrial cancer cell signaling pathways （ MAPK/ERK, PI3 K/Akt ） regulation. Methods： Epidermal growth factor HER -2/neu transfected Ishikawa cell line before and after, Western -blot transfected cells before and after the expression of COX - 2, t - Akt and p - Akt protein, t - ERK and p - ERK, ELISA method for the detection of cell culture supernatants of estradiol contents. PI3K/Akt inhibitor LY294002 and MAPK/ERK inhibitor PD98059 inhibited signaling pathways, with the same concentration at different time and different concentration to detec COX -2 and estradiol levels. Results： The HER - 2/neu transfected Ishikawa cell line COX - 2 protein, protein p - Akt ; p - ERK and cell supernatant in estradiol content were higher than the normal Ishikawa cell line （ P 〈 0.05 ）. Inhibitor in- hibits two pathways, transfected cells the expression level of COX -2 in transfected Ishikawa cell line than before, at the same time E2 in the transfeeted group was compared with non transfected group decreased obviously. With the increase of the concentration and action time, two groups of cells COX - 2 and estradiol expression were gradually decreased, and the inhibitory action and concentration and action time were negatively related. Conclusion： Endometrial carcinoma cell line Ishikawa HER -2/neu enhanced gene expression induced by COX -2,increase the expression of E2.
出处
《现代肿瘤医学》
CAS
2013年第2期242-247,共6页
Journal of Modern Oncology
基金
辽宁省自然科学基金资助项目(编号:20102266)
"百千万人才工程"百人层次项目(编号:2010921071)
辽宁省教育厅科研项目(编号:2010642)
