摘要
目的:探究表没食子儿茶素没食子酸酯(epigallocatechin gallate,EGCg)特异性抑制芳香乙酰胺脱乙酰基酶蛋白(arylacetamide deacetylase,AADAC)来诱导卵巢癌细胞发生铁死亡的分子机制。方法:体外培养卵巢癌细胞系(SK-OV-3、A2780、Hey和SW626)。CCK-8检测EGCg对卵巢癌细胞的半数抑制浓度(median inhibition concentration,IC_(50))。铁离子检测试剂盒检测细胞内Fe^(2+)的含量。活性氧(reactive oxygen species,ROS)检测试剂盒检测细胞内ROS的含量。Western blot检测细胞内铁死亡相关蛋白GPX4和NOX1的表达。酶活实验检测EGCg对AADAC活性的影响。Western blot检测EGCg对AADAC蛋白的影响。应用免疫组化和Western blot检测卵巢癌患者组织中AADAC的表达。应用siRNA沉默卵巢癌细胞总AADAC的表达。应用AADAC-KO敲除卵巢癌细胞中AADAC的表达。皮下成瘤验证小鼠体内EGCg对卵巢癌移植瘤生长的影响。结果:EGCg能抑制卵巢癌细胞的增殖,且对SK-OV-3和SW626细胞较为敏感。EGCg使卵巢癌细胞中Fe^(2+)含量增加,ROS含量增加,并抑制GPX4蛋白表达,促进NOX1蛋白的表达。EGCg能抑制AADAC蛋白的活性并促进其降解。AADAC在卵巢癌患者癌组织中高表达。沉默AADAC能促进卵巢癌细胞发生铁死亡。沉默AADAC后EGCg对卵巢癌细胞变得不敏感,并不能促进铁死亡的发生。相对于NC组,EGCg能抑制裸鼠移植瘤的生长,且移植瘤中AADAC表达下降,铁死亡被激活。结论:EGCg能特异性抑制AADAC来诱导卵巢癌细胞发生铁死亡。
Objective:To explore the molecular mechanism of epigallocatechin gallate(EGCg)specifically inhibiting arylacetamide deacetylase(AADAC)to induce ferroptosis in ovarian cancer cells.Methods:Ovarian cancer cell lines(SK-OV-3,A2780,Hey and SW626)were cultured in vitro.CCK-8 was used to detect median inhibition concentration(IC_(50))of EGCg in ovarian cancer cells.The iron ion detection kit detected the content of Fe^(2+)ions in cells.The reactive oxygen species(ROS)detection kits measured the amount of ROS in cells.The expressions of ferroptosis related proteins GPX4 and NOX1 were detected by Western blot.The effect of EGCg on AADAC activity was detected by enzyme activity test.The effect of EGCg on AADAC protein was detected by Western blot.Immunohistochemistry and Western blot were used to detect AADAC expression in ovarian cancer patients.siRNA was used to silence the expression of total AADAC in ovarian cancer cells.AADAC expression in ovarian cancer cells was knocked down by AADAC-KO.The effect of EGCg on the growth of ovarian cancer transplantation tumor was verified by subcutaneous tumorigenesis.Results:EGCg could inhibit the proliferation of ovarian cancer cells and is sensitive to SK-OV-3 and SW626 cells.EGCg increased the content of Fe^(2+)and ROS in ovarian cancer cells,inhibited the expression of GPX4 protein and promoted the expression of NOX1 protein.EGCg could inhibit the activity and degradation of AADAC protein.AADAC was highly expressed in ovarian cancer patients.Silencing AADAC promoted ferroptosis in ovarian cancer cells.EGCg became insensitive to ovarian cancer cells after knockout of AADAC and did not promote ferroptosis.Compared with NC group,EGCg inhibited the growth of transplanted tumor in nude mice,and AADAC expression was decreased and ferroptosis was activated in transplanted tumor.Conclusion:EGCg could specifically inhibit AADAC to induce ferroptosis in ovarian cancer cells.
作者
蔡月红
麦燕
钱沁佳
CAI Yuehong;MAI Yan;QIAN Qinjia(Department of Obstetrics and Gynecology,Sanya Central Hospital,Hainan Sanya 572000,China)
出处
《现代肿瘤医学》
CAS
北大核心
2023年第23期4326-4333,共8页
Journal of Modern Oncology
基金
海南省卫生计生行业科研计划项目(编号:18A200164)。
