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目的通过观察促卵泡激素(FSH)、活性氧(ROS)及其特异性抑制剂对卵巢癌细胞中红系衍生的核因子2相关因子2(nuclear factor-erythroid 2-related factor-2,Nrf2)表达的影响,探讨FSH是否通过ROS途径调控卵巢癌细胞Nrf2的表达。方法 (1)采用免疫组织化学方法检测Nrf2在卵巢癌良恶性组织中的表达。(2)用不同浓度的FSH刺激卵巢癌Hey细胞不同时间,采用蛋白质印迹法检测细胞内Nrf2蛋白的表达。(3)采用ROS检测试剂盒检测80 mIU/ml的FSH刺激后不同时间Hey细胞内ROS的生成情况。(4)Hey细胞经不同浓度的H2O2刺激48 h后,或经ROS特异性抑制剂N-乙酰半胱氨酸(NAC)预处理后加入150 mmol/L的H2O2处理48 h,观察细胞内Nrf2的表达。(5)Hey细胞经不同浓度的NAC预处理1 h,加入80 mIU/ml的FSH共孵育48 h,观察细胞内Nrf2表达的变化。结果 (1)10例良性卵巢囊肿组织中仅3例有Nrf2蛋白的弱阳性表达,而64例卵巢癌组织中42例(65.6%)有Nrf2蛋白的阳性表达,两组差异有统计学意义(P=0.042)。(2)FSH可上调Hey细胞内Nrf2的表达,并呈一定的剂量、时间依赖效应。(3)FSH可促进Hey细胞ROS的产生。(4)H2O2可诱导Hey细胞Nrf2蛋白的表达,且其作用可被NAC阻断。(5)NAC可阻断FSH所诱导的Nrf2蛋白的表达。结论 FSH可促进卵巢癌Hey细胞中ROS产生,上调Nrf2蛋白表达,抑制ROS途径可阻断FSH诱导的Nrf2高表达。提示FSH可能通过ROS途径调控卵巢癌Nrf2的表达,进而促进肿瘤发生。
Objective To observe the effects of erythroid 2-related factor-2 (Nrf2) and erythroid-derived factor-2 (NF-κB) in ovarian cancer cells by observing the effects of follicle stimulating hormone (FSH), reactive oxygen species (ROS) and their specific inhibitors. To investigate whether FSH regulates the expression of Nrf2 in ovarian cancer cells through ROS pathway. Methods (1) Immunohistochemistry was used to detect the expression of Nrf2 in benign and malignant ovarian tissues. (2) Different concentrations of FSH stimulated Hey cells of ovarian cancer cells at different times, Western blotting was used to detect the expression of Nrf2 protein. (3) The production of ROS in Hey cells was detected by ROS assay kit at 80 mIU / ml FSH. (4) Hey cells were stimulated with H2O2 at different concentrations for 48 h or pretreated with NAC (NAC) and then treated with 150 mmol / L H2O2 for 48 h. The levels of Nrf2 expression. (5) Hey cells were pretreated with different concentrations of NAC for 1 h and incubated with 80 mIU / ml FSH for 48 h. The changes of Nrf2 expression were observed. Results (1) Nrf2 protein expression was weakly positive in only 3 of 10 benign ovarian cysts and 42 (65.6%) of 64 ovarian cancers were positive for Nrf2 protein, with significant difference between the two groups P = 0.042). (2) FSH up-regulated the expression of Nrf2 in Hey cells in a dose-and time-dependent manner. (3) FSH can promote Hey cell ROS production. (4) H2O2 can induce the expression of Nrf2 protein in Hey cells, and its effect can be blocked by NAC. (5) NAC can block FSH induced Nrf2 protein expression. Conclusion FSH can promote the production of ROS in Hey cells of ovarian cancer, upregulate the expression of Nrf2 protein and inhibit the ROS pathway to block the FSH-induced high expression of Nrf2. It is suggested that FSH might regulate the expression of Nrf2 in ovarian cancer through ROS pathway, and then promote tumorigenesis.