摘要
目的:探讨槐耳多糖诱导三阴性乳腺癌MDA-MB-231细胞铁死亡的作用机制。方法:通过CCK-8实验检测槐耳多糖不同浓度、不同干预时间对MDA-MB-231细胞活性的影响;取对数生长期的MDA-MB-231细胞,随机分成6组,分别为对照组(NC组)、槐耳多糖组(PS-T组)、槐耳多糖+铁诱导剂组(PS-T+Erastin组)、铁诱导剂组(Erastin组)、紫杉醇组(PTX组)和槐耳多糖+紫杉醇组(PS-T+PTX组),用克隆形成实验检测各组MDA-MB-231细胞的增殖情况;用铁死亡相关试剂盒检测细胞内亚铁离子(Fe^(2+))、谷胱甘肽(GSH)和丙二醛(MDA)的含量;通过JC-1线粒体膜电位检测试剂盒检测MDA-MB-231细胞的线粒体膜电位,并用流式细胞仪进行分析;用DCFH-DA探针对活性氧(ROS)进行检测,并用荧光显微镜进行分析;用实时定量PCR仪检测MDA-MB-231细胞内p53 mRNA、SLC7A11 mRNA、GPX4 mRNA、TFR1 mRNA和DMT1 mRNA的表达情况。结果:与NC组比较,乳腺癌细胞的活力随槐耳多糖的浓度升高而逐渐降低(P<0.05),且24 h比48 h的作用更显著;与NC组比较,PS-T组、PS-T+Erastin组、Erastin组、PTX组和PS-T+PTX组的克隆形成率、GSH含量、线粒体膜电位以及GPX4 mRNA和SLC7A11 mRNA表达均显著下降,而Fe^(2+)、MDA含量、ROS水平以及DMT1 mRNA、TFR1 mRNA和p53 mRNA表达则显著升高(P<0.05);与PS-T组比较,PS-T+Erastin组和PS-T+PTX组的克隆形成率、GSH含量、线粒体膜电位以及GPX4 mRNA和SLC7A11 mRNA表达均下调,而Fe^(2+)、MDA含量、ROS水平以及DMT1 mRNA、TFR1 mRNA和p53 mRNA表达则上调(P<0.05)。结论:槐耳多糖可以诱导三阴性乳腺癌MDA-MB-231细胞的铁死亡,其作用机制可能与Fe^(2+)积累和脂质过氧化相关。
Objective:To investigate the mechanism of ferroptosis induced by sophora polysaccharides in triple-negative breast cancer MDA-MB-231 cells.Methods:CCK-8 assay was used to detect the effects of different concentrations and different times of polysaccharides on the activity of MDA-MB-231 cells.The MDA-MB-231 cells in logarithmic growth phase were randomly divided into 6 groups,including control group(NC group),sophora polysaccharide group(PS-T group),sophora polysaccharide+iron inducion dose group(PS-T+Erastin group),iron inducer group(Erastin group),paclitaxel group(PTX group)and sophora polysaccharide+paclitaxel group(PS-T+PTX group).The proliferation of MDA-MB-231 cells in each group was detected by clonal formation assay.The contents of ferrous ions(Fe^(2+)),glutathione(GSH)and malondialdehyde(MDA)were detected with ferroptosis-related kits.The mitochondrial membrane potential of MDA-MB-231 cells was detected by JC-1 mitochondrial membrane potential detection kit and analyzed by flow cytometry.DCFH-DA probe was used to detect reactive oxygen(ROS),and fluorescence microscope was used for analysis.The p53 mRNA,SLC7A11 mRNA,GPX4 mRNA,TFR1 mRNA and DMT1 mRNA levels of MDA-MB-231 were detected by real-time quantitative PCR instrument.Results:Compared with NC group,the viability of breast cancer cells decreased gradually with the increase of the concentration of sophora polysaccharides(P<0.05),and the effect was more significant in 24 h than in 48 h.Compared with NC group,the clone formation rate,GSH content,mitochondrial membrane potential,GPX4 mRNA expression and SLC7A11 mRNA expression decreased in PS-T group,PS-T+Erastin group,Erastin group,PTX group and PS-T+PTX group,while Fe^(2+),MDA content,ROS level,DMT1 mRNA expression,TFR1 mRNA expression and p53 mRNA expression were significantly increased(P<0.05).Compared with PS-T group,the clonogenic rate,GSH content,mitochondrial membrane potential,GPX4 mRNA expressions and SLC7A11 mRNA expressions were down-regulated in PS-T+Erastin grouop and PS-T+PTX group,while Fe^(2+),MDA content,ROS levels,DMT1 mRNA expression,TFR1 mRNA expression,and p53 mRNA expression were up-regulated(P<0.05).Conclusion:Sophora polysaccharides can induce ferroptosis in triple negative breast cancer MDA-MB-231 cells,and its mechanism may be related to Fe^(2+)accumulation and lipid peroxidation.
作者
王玲
高翔
高爽
汪唐顺
冯雪
王玉坤
孙萍
左禧萌
史晓光
WANG Ling;GAO Xiang;GAO Shuang;WANG Tangshun;FENG Xue;WANG Yukun;SUN Ping;ZUO Ximeng;SHI Xiaoguang(Dongzhimen Hospital,Beijing University of Chinese Medicine,Beijing 100700,China)
出处
《中医药导报》
2025年第8期47-52,共6页
Guiding Journal of Traditional Chinese Medicine and Pharmacy
基金
国家自然科学基金项目(82474510)
湖北陈孝平科技发展基金会专项基金项目(CXPJJH12000002-2020010)
北京中医药大学青年教师项目(2022-BUCMXJKY009)。
关键词
乳腺恶性肿瘤
槐耳多糖
铁死亡
线粒体膜电位
脂质过氧化
malignant tumors of the breast
sophora polysaccharides
ferroptosis
mitochondrial membrane potential
lipid peroxidation
