上海交通大学学报（医学版）2025，Vol.45Issue(10)：1298-1307,10.DOI:10.3969/j.issn.1674-8115.2025.10.005
"HA糖外衣"调控ER+乳腺癌细胞干性在内分泌治疗耐药中的作用
Role of"HA coat"in modulating stemness and endocrine resistance in ER+breast cancer
摘要
Abstract
Objective·To determine hyaluronan(HA)expression in the endocrine-resistant microenvironment of estrogen receptor-positive(ER+)breast cancer and elucidate its impact on the acquired resistance.Methods·Chemiluminescent immunoassay was used to quantify HA levels in the culture supernatants of fulvestrant-resistant breast cancer cells.An immunofluorescence(IF)assay was performed to visualize the colocalization of CD44 and HA in MCF7/FulR cells.Using an established adaptive endocrine-resistant breast cancer mouse model,HA expression in resistant breast cancer tissues was assessed by immunohistochemistry(IHC) assay.Single-cell RNA sequencing(scRNA-seq)and RNA sequencing(RNA-seq)were conducted to examine transcriptomic profiles and alterations in HA-related genes in resistant breast cancer cells.Flow cytometry(FCM)was utilized to measure the proportion of CD44+CD24-cells in MCF7/FulR.The correlation between HA synthesis genes and cell stemness was investigated in clinical ER+breast cancers from GEO data sets.Hyaluronidase(HAase)treatment was applied to remove the"HA coat",and RT-qPCR and Western blotting analysis were carried out to monitor changes in stemness-related molecules.CCK-8 assays,flow cytometry(FCM),and Hoechst 33258 staining were performed to determine changes in apoptosis and fulvestrant efficiency after HAase treatment.Results·IF results revealed that compared with MCF7 cells,the"HA coat"on the surface of MCF7/FulR cells was significantly thickened.IHC demonstrated markedly increased HA retention in fulvestrant-resistant mouse breast cancer tissues.ScRNA-seq and RNA-seq analyses indicated elevated expression of stemness-related genes and HA synthesis-associated genes in fulvestrant-resistant breast cancer cells.Correlation analysis revealed a positive association between HA synthesis and cancer stemness in ER+breast cancer.IF and RT-qPCR results demonstrated that removing the HA coating from the surface of MCF7/FulR cells led to a significant reduction in the expression of stemness-related molecules;concurrently,CCK-8 assays,FCM analysis,and Hoechst 33258 staining revealed that"HA coat"clearance reduced MCF7/FulR'tolerance to fulvestrant and increased apoptosis.Conclusion·Endocrine-resistant breast cancer cells develop an enriched"HA coat",which promotes stemness in fulvestrant-resistant tumors.Disruption of this HA coat through HAase treatment effectively reduces cell stemness,induces apoptosis,and re-sensitizes breast cancer cells to fulvestrant.关键词
乳腺癌/内分泌治疗耐药/透明质酸糖外衣Key words
breast cancer/endocrine therapy-resistant tumor/hyaluronan coat分类
医药卫生引用本文复制引用
吴诗怡,高锋,杨翠霞,陈思,刘泊含,刘宇婷,刘鷖雯,何怡青,杜艳,张国良,郭倩.."HA糖外衣"调控ER+乳腺癌细胞干性在内分泌治疗耐药中的作用[J].上海交通大学学报（医学版）,2025,45(10):1298-1307,10.基金项目
国家自然科学基金(82273462,81974445,82503431). National Natural Science Foundation of China(82273462,81974445,82503431). （82273462,81974445,82503431）
