MUC1介导三阴性乳腺癌细胞铁稳态失衡及槲皮素的干预作用  

MUC1 Mediates Iron Metabolism Disorder in Triple-Negative Breast Cancer Cells and Quercetin Intervention

作　　者：王海娇 谢伍星 徐蓉 梅志刚 刘永平 陈懿 WANG Hai-jiao;XIE Wu-xing;XU Rong;MEI Zhi-gang;LIU Yong-ping;CHEN Yi(College of Integrative Medicine,Hunan University of Traditional Chinese Medicine,Changsha 410208,China;College of Medicine,Hunan University of Traditional Chinese Medicine,Changsha 410208,China)

机构地区：[1]湖南中医药大学中西医结合学院,长沙410208 [2]湖南中医药大学医学院,长沙410208

出　　处：《湖南师范大学自然科学学报》2024年第6期123-131,共9页Journal of Natural Science of Hunan Normal University

基　　金：湖南省教育厅重点项目(23A0301);湖南中医药大学中西医结合一流学科重点项目(2021ZXYJH02);湖南中医药大学研究生创新课题(2023CX158)。

摘　　要：为了研究人粘液蛋白1(MUC1)对三阴性乳腺癌(TNBC)铁稳态的影响,探讨槲皮素抗TNBC增殖的新机制,采用生物信息学方法分析人正常乳腺组织和乳腺癌组织中MUC1的表达及与预后的相关性;采用慢病毒转染构建MUC1过表达细胞模型,将三阴性乳腺癌MDA-MB-468细胞分成对照组、不同浓度槲皮素组和GO203(MUC1抑制剂)组;通过CCK-8法检测细胞活力,亚铁离子荧光探针检测细胞内游离Fe^(2+)浓度,qPCR方法检测MUC1基因表达,Western Blot方法检测MUC1及铁代谢相关蛋白表达。研究结果表明,MUC1在乳腺癌组织的表达远高于正常乳腺组织(P<0.05),TNBC患者预后与MUC1高表达密切相关(P<0.01);与对照组相比,MUC1过表达模型细胞增殖明显加快(P<0.01),细胞内Fe^(2+)浓度增加(P<0.01),铁输入蛋白TFR1和FTH1表达增加(P<0.05,P<0.05),但铁输出蛋白SLC40A1、铁自噬调节因子NCOA4及上游调节因子NRF2表达无统计学变化(P>0.05);槲皮素可降低MUC1的表达(P<0.01,P<0.05),槲皮素和MUC1抑制剂G0203均可显著抑制TNBC细胞增殖(P<0.05),减少细胞内铁含量(P<0.01),降低铁输入蛋白TFR1和FTH1的表达(P<0.01,P<0.05),增加铁输出蛋白SLC40A1的表达(P<0.05)。综上可得,高表达MUC1可影响铁代谢相关蛋白表达,导致铁稳态失衡,进而铁积累促进细胞增殖;槲皮素可显著抑制MDA-MB-468细胞增殖,其机制可能是通过下调MUC1调节铁稳态,降低细胞内铁水平。In order to study the effect of human mucin 1(MUC1)on the iron homeostasis in triple-negative breast cancer(TNBC)and investigate the new mechanism of quercetin against the TNBC proliferation,the expression of MUC1 in the normal breast tissue and breast cancer tissue along with its correlation with prognosis were analyzed by the bioinformatics method in this study.The MUC1 overexpression cell model was constructed using lentivirus transfection,and the triple-negative breast cancer MDA-MB-468 cells were divided into a control group,a quercetin group with different concentrations,and a GO203(MUC1 inhibitor)group.The cell viability was detected by the CCK-8 method,the level of free Fe^(2+)was detected by a fluorescence probe of ferrous ion,the expression of MUC1 gene was detected by qPCR,and the expression of MUC1 and iron metabolism-related proteins was detected by Western Blot.The results showed that the expression of MUC1 in the breast cancer tissue was much higher than that in the normal breast tissue(P<0.05),and the prognosis of TNBC patients was closely related to the high expression of MUC1(P<0.01).Compared with the control group,the proliferation of MUC1 over-expression model cells was significantly accelerated(P<0.01),accompanied by an increase in the intracellular Fe^(2+)level(P<0.01)and also increases in the expressions of ferric input protein TFR1 and FTH1(P<0.05,P<0.05).However,there were no significant changes in the expressions of iron exporting protein SLC40A1,ferritinophagy regulatory factor NCOA4 and upstream regulatory factor NRF2(P>0.05).Quercetin could reduce the MUC1 gene and protein expressions(P<0.01,P<0.05);quercetin and G0203(MUC1 inhibitors)could inhibit the TNBC cell proliferation(P<0.05),reduce the iron content in cells(P<0.01),lower the iron input TFR1 protein and FTH1 expressions(P<0.01,P<0.05),and increase the expression of iron exporting protein SLC40A1(P<0.05).In conclusion,the high expression of MUC1 can affect the expression of iron metabolism-related proteins,leading to an iron ho

关 键 词：槲皮素 三阴性乳腺癌 MUC1 铁稳态 抗肿瘤 

分 类 号：R737.9[医药卫生—肿瘤]