毛大丁草内酯J对乳腺癌细胞周期和凋亡的影响及机制研究  

Effect and mechanism of gerbeloid J from Gerbera piloselloides(L.)Cass.on cycle and apoptosis of breast cancer cells

作　　者：李靖荣 李凌宇[2] 赵晨旭 尚海[2] 张涛[2] 邹忠梅[2] 宛蕾 LI Jing-rong;LI Ling-yu;ZHAO Chen-xu;SHANG Hai;ZHANG Tao;ZOU Zhong-mei;WAN Lei(School of Basic Medical Sciences,Guizhou Medical University,Guiyang 550025,China;Institute of Medicinal Plant Development,Chinese Academy of Medical Sciences and Peking Union College,Beijing 100193,China)

机构地区：[1]贵州医科大学基础医学院,贵州贵阳550025 [2]中国医学科学院、北京协和医学院,药用植物研究所,北京100193

出　　处：《药学学报》2023年第4期938-945,共8页Acta Pharmaceutica Sinica

基　　金：中国医学科学院医学与健康科技创新工程(2022-I2M-1-017)。

摘　　要：乳腺癌已成为女性发病率最高的恶性肿瘤,严重危害女性的健康。从传统药物中筛选出高效低毒的先导化合物是开发乳腺癌治疗药物的有效途径之一。毛大丁草内酯J是本课题组前期从民族药毛大丁草中分离得到的一种新型香豆素类化合物,具有显著的抗肿瘤活性,但关于毛大丁草内酯J对乳腺癌细胞周期和凋亡的影响及作用机制尚不明确。本研究利用CCK-8法、克隆形成实验、PI染色法探讨毛大丁草内酯J对乳腺癌细胞MCF-7和MDA-MB-231细胞增殖及周期的影响。通过DAPI、Annexin V/TO-PRO-3、Rhod-2 AM、TMRM、DCFDA染色法及蛋白免疫印迹法考察毛大丁草内酯J对MCF-7和MDA-MB-231细胞凋亡及线粒体功能的影响。结果表明,毛大丁草内酯J通过调控P21/CDC25C/CDK-1/cyclin B1通路,将细胞周期阻滞于G2/M期,显著抑制MCF-7和MDA-MB-231细胞增殖。此外,毛大丁草内酯J能诱导MCF-7和MDA-MB-231细胞发生线粒体钙超载,线粒体膜电位降低,活性氧生成增加,引发细胞凋亡,其作用机制与激活线粒体凋亡通路有关。综上,毛大丁草内酯J可通过调控P21/CDC25C/CDK-1/cyclin B1通路及激活线粒体凋亡通路诱导乳腺癌细胞周期阻滞及凋亡,为乳腺癌新药研发提供新的候选化合物。Breast cancer has become the most prevalent malignant tumor among women,putting the health of women at serious risk.Screening for lead compounds in the active ingredients of plant that are effective and less toxic continues to be an important strategy for treating breast cancer.Gerbeloid J,a coumarin isolated from Gerbera piloselloides(L.)Cass.,showed significant anti-cancer activity.But there is no report on the effect and mechanism of gerbeloid J on cycle and apoptosis of breast cancer.By using the CCK-8,clone formation,and PI staining assays,the effects of gerbeloid J on the proliferation of MCF-7 and MDA-MB-231 cells were assessed in this study.The effects of gerbeloid J on the apoptosis and mitochondrial function of MCF-7 and MDA-MB-231 cells were assessed using DAPI,Annexin V/TO-PRO-3,Rhod-2 AM,TMRM,DCFDA staining assays,and Western blot.The results demonstrated that gerbeloid J regulated the P21/CDC25C/CDK-1/cyclin B1 pathway and arrested the cell cycle at G2/M phase to suppressed the proliferation of MCF-7 and MDA-MB-231 cells.Additionally,gerbeloid J induced apoptosis through the stimulation of mitochondrial calcium excess,reduction of mitochondrial membrane potential,and promotion of ROS generation,and its mechanism was related to the activation of mitochondrial apoptotic pathway.In conclusion,by regulating the P21/CDC25C/CDK-1/cyclin B1 pathway and activating the mitochondrial apoptosis pathway,gerbeloid J could cause breast cancer cell cycle arrest and apoptosis,which might offer a promising candidate for the creation of new drugs against breast cancer.

关 键 词：乳腺癌 毛大丁草 毛大丁草内酯J 增殖 凋亡 

分 类 号：R966[医药卫生—药理学]