二氢杨梅素对顺铂体外抗乳腺癌作用的干预及其机制研究  

Dihydromyricetin Enhances the Cytotoxicity of Cisplatin to Breast Cancer Through Inhibiting the SIRT1/ROS/JNK Pathway

作　　者：胡蓉蓉 徐广涛[2] 

机构地区：[1]浙江省嘉兴市中医院检验科,嘉兴314000 [2]浙江省嘉兴学院医学院检验科,嘉兴314000

出　　处：《浙江中西医结合杂志》2018年第3期184-187,共4页Zhejiang Journal of Integrated Traditional Chinese and Western Medicine

摘　　要：目的观察二氢杨梅素对顺铂体外抗乳腺癌作用的影响,并探讨其作用机制。方法将2μg/m L沉默信息调节因子2相关酶1(SIRT1)表达质粒转染MDA-MB-435细胞,MDA-MB-435细胞按5×103/孔接种在96孔板上,分为对照组、二氢杨梅素组、顺铂组、顺铂+二氢杨梅素组、顺铂+二氢杨梅素+NAC组和顺铂+二氢杨梅素+SIRT1质粒组。分别采用MTT法和Annexin V染色流式细胞术检测MDA-MB-435细胞活力和细胞凋亡。Western blot方法检测MCF-10A及MCF-7、MDA-MB-231、MDA-MB-435细胞SIRT1表达水平。二氢乙啶(DHE)染色流式细胞术及Western blot方法分别检测ROS产生和JNK磷酸化。结果二氢杨梅素联合顺铂处理的MDA-MB-435细胞活力抑制率和凋亡诱导率显著高于顺铂组、二氢杨梅素组和二氢杨梅素+顺铂+SIRT1质粒组[细胞活力抑制率:(61.5±5.1)%比(15.2±1.4)%、(7.2±0.5)%、(19.7±1.6)%,P均<0.05;凋亡诱导率:(36.8±2.9)%比(7.6±0.6)%、(2.9±0.3)%、(10.8±0.9)%,P均<0.05]。人乳腺癌细胞系MCF-7、MDAMB-231和MD-MB-435SIRT1蛋白表达水平明显高于人正常乳腺上皮细胞系MCF-10A。二氢杨梅素处理显著抑制MDA-MB-435细胞SIRT1蛋白表达水平并显著增强顺铂对MDA-MB-435细胞活性氧簇(ROS)的诱导、JNK蛋白磷酸化和Caspase-9、Caspase-3活化。结论二氢杨梅素可能通过抑制SIRT1表达增强顺铂对乳腺癌细胞的杀伤活性。Objective To investigate the synergistic effect and mechanism of dihydromyricetin on enhancing cisplatin-induced cytotoxicity to breast cancer. Methods MDA-MB-435 cells were transfected with SIRT1 plasmid of 2μg/m L, then divided into control group, dihydromyricetin group, cisplatin group, cisplatin +dihydromyricetin group,cisplatin+dihydromyricetin+NAC group, and cisplatin+dihydromyricetin+SIRT1 plasmid group. MTT assay and Annexin V staining were performed to investigate the cell viability and apoptosis of MDA-MB-435 cells in each group.Western blot analysis was performed to determine the expression level of SIRT1 in MCF-10 A and breast cancer cell lines MCF-7, MDA-MB-231 and MDA-MB-435. DHE staining and western blot analysis were performed to detect the production of ROS and phosphorylation of JNK. Results Cell viability inhibitory rate(61.5%±5.1%) and apoptotic rate(36.8%±2.9%) of MDA-MB-435 cells co-treated with dihydromyricetin and cisplatin was significantly higher than the cell viability inhibitory rate(15.2%±1.4%) and apoptotic rate(7.6%±0.6%) in the cisplatin single treatment group, and the cell viability inhibitory rate(19.7%±1.6%) and apoptotic rate(10.8%±0.95) in the cisplatin+dihydromyricetin +SIRT1 plasmid group. The expression of SIRT1 was obviously upregulated in breast cancer cell lines MCF-7, MDA-MB-231 and MDA-MB-435 rather than the normal mammary epithelial cell line MCF-10 A.Dihydromyricetin significantly inhibited the expression of SIRT1 followed by enhancing the cisplatin-induced ROS production, JNK phosphorylation and activation of caspase-9 and caspase-3 in MDA-MB-435 cells. Conclusion Dihydromyricetin enhances the cytotoxicity of cisplatin to breast cancer through inhibiting the expression of SIRT1.

关 键 词：乳腺癌 MCF-10A MDA-MB-435 SIRT1 ROS/JNK 凋亡 二氢杨梅素 顺铂 

分 类 号：R737.9[医药卫生—肿瘤]