藤黄酸抑制人卵巢透明细胞癌ES-2细胞增殖和促进其凋亡的机制研究  

Mechanism of gambogic acid inhibiting proliferation and promoting apoptosis of human ovarian clear cell carcinoma ES-2 cells

作　　者：殷敏 陈志高 周慧梅[1] 杨佳欣[1] YIN Min;CHEN Zhigao;ZHOU Huimei;YANG Jiaxin(Department of Obstetrics and Gynecology,Peking Union Medical College Hospital,Peking Union Medical College,Chinese Academy of Medical Sciences/National Clinical Research Center for Obstetrics and Gynecology,Beijing,100730,China;Institute of Basic Medical Sciences,Peking Union Medical College,Chinese Academy of Medical Sciences,Beijing,100730,China)

机构地区：[1]中国医学科学院北京协和医学院北京协和医院妇产科/国家妇产疾病临床医学研究中心,北京100730 [2]中国医学科学院北京协和医学院基础医学研究所,北京100730

出　　处：《肿瘤药学》2022年第4期465-471,共7页Anti-Tumor Pharmacy

基　　金：中国医学科学院医学与健康科技创新工程(2017-I2M-1-002)。

摘　　要：目的研究藤黄酸对人卵巢透明细胞癌ES-2细胞增殖和凋亡的影响及其潜在分子机制。方法采用CCK-8检测藤黄酸对不同类型人卵巢癌细胞增殖的影响;克隆形成实验检测藤黄酸对ES-2细胞克隆形成的影响;细胞划痕愈合实验及Transwell实验分别检测藤黄酸对ES-2细胞迁移与侵袭的影响;Caspase-Glo 3/7试剂盒检测藤黄酸对ES-2细胞内Caspase 3/7活性的影响;Annexin V-FITC/PI双染色法分析藤黄酸对ES-2细胞凋亡的影响;Western blotting检测不同浓度藤黄酸对ES-2细胞中Bcl-2、Bax、PTEN、PI3K、p-AKT、p-mTOR水平的影响。结果人卵巢癌ES-2、HO8910、SKOV3、OVCAR3细胞中,藤黄酸对卵巢透明细胞癌ES-2细胞的增殖抑制作用最明显,且随着藤黄酸浓度和作用时间的增加而增强;藤黄酸可抑制ES-2细胞的克隆形成、迁移及侵袭、增强Caspase3/7活性并促进细胞凋亡,且呈浓度依赖性;藤黄酸可下调ES-2细胞中Bcl-2、PI3K、p-AKT及p-mTOR的表达,上调Bax、PTEN的表达,且呈浓度依赖性。结论藤黄酸能够抑制卵巢透明细胞癌ES-2细胞的增殖、迁移和侵袭,并诱导其凋亡,其作用机制可能与调控PI3K/AKT/mTOR信号通路相关。Objective To study the effects of gambogic acid on the proliferation and apoptosis of human ovarian clear cell carcinoma ES-2 cells and its underlying molecular mechanism.Methods The effects of gambogic acid on the proliferation of different types of ovarian cancer cells was detected by CCK-8 method.The effects of gambogic acid on the colony formation of ES-2 cells was detected by colony formation assay.The effects of gambogic acid on the migration and invasion ability of ES-2 cells was detected by cell scratch healing assay and transwell assay respectively.Caspase-Glo 3/7 kit was used to detect the activity of caspase 3/7 in ES-2 cells after treatment with gambogic acid.Annexin V-FITC/PI double staining method was used to analyze the effects of gambogic acid on the apoptosis of ES-2 cells.Western blotting was used to detect the changes in the expression levels of Bcl-2,Bax,PTEN,PI3K,p-AKT and p-mTOR in ES-2 cells.Results Among the four human ovarian cancer cell lines,ES-2,HO8910,SKOV3,and OVCAR3,gambogic acid had the most obvious inhibitory effects on the proliferation of ES-2 cells.And it inhibited the proliferation of ES-2 cells in a time-and concentration-dependent way.Gambogic acid also inhibited the colony formation,migration and invasion of ES-2 cells in a concentration-dependent manner.Additionally,gambogic acid enhanced the activity of caspase 3/7 and promoted cell apoptosis in a concentration-dependent manner.The expression levels of Bcl-2,PI3K,p-AKT and p-mTOR were decreased,while the expression levels of Bax and PTEN were increased in a concentration-dependent manner after gambogic acid treatment in ES-2 cells.Conclusion Gambogic acid can inhibit the proliferation,migration and invasion of ovarian clear cell carcinoma ES-2 cells and induce their apoptosis.Its mechanism may be related to the regulation of PI3K/AKT/mTOR signaling pathway.

关 键 词：藤黄酸 卵巢癌 卵巢透明细胞癌 PI3K/AKT/MTOR 

分 类 号：R737.31[医药卫生—肿瘤]