癌症干细胞（CSC）导致乳腺癌药物复发和耐药，从而导致死亡和治疗失败。在这项研究中，我们检查了暴露于低浓度阿霉素 (Dox) 对来自 TNBC 的 CSC 和非 CSC 的影响。使用 SORE6 报告系统研究了 Dox 的影响。我们通过流式细胞术检查了 CSC 群体的富集，以及报告基因阳性部分（GFP  细胞）的增殖和死亡。分别通过活力和微球形成测定来分析抗性和干性表型。我们通过 RNA-seq 分析确定了差异表达和共调控的基因，并使用公共数据库通过 Kaplan-Mayer 分析评估了基因表达与临床结果之间的相关性。在 MDAMB231 和 Hs578t 细胞中，我们在连续暴露后确定了 CSC 群体中的富集子集低浓度的 Dox。来自这些富集培养物的细胞表现出对有毒浓度的 Dox 的抵抗力，并提高了微球形成的效率。在纯化的 GFP 或 GFP- 细胞中，Dox 提高了乳腺球形成效率，促进非 CSC 群体的表型转变为 CSC 样状态，减少增殖并诱导差异基因表达。我们确定了几种生物过程和分子功能，部分解释了阿霉素耐药细胞的发育和细胞可塑性。在受 Dox 暴露调节的基因中，ITGB1、SNAI1、NOTCH4、STAT5B、RAPGEF3、LAMA2 和 GNAI1 的表达与较差的存活率、干性表型和化疗耐药性显着相关。具有以下特征的化疗耐药细胞的产生CSC 在暴露于低浓度的 Dox 后，涉及具有临床影响的基因的差异表达。© 2024。作者。

Cancer stem cells (CSCs) account for recurrence and resistance to breast cancer drugs, rendering them a cause of mortality and therapeutic failure. In this study, we examined the effects of exposure to low concentrations of doxorubicin (Dox) on CSCs and non-CSCs from TNBC.The effects of Dox were studied using the SORE6 reporter system. We examined the enrichment of the CSCs population, as well as the proliferation, and death of the reporter-positive fraction (GFP + cells) by flow cytometry. The resistant and stemness phenotypes were analyzed by viability and mammosphere formation assay, respectively. We identified differentially expressed and coregulated genes by RNA-seq analysis, and the correlation between gene expression and clinical outcome was evaluated by Kaplan-Mayer analysis using public databases.In MDAMB231 and Hs578t cells, we identified enriched subsets in the CSCs population after continuous exposure to low concentrations of Dox. Cells from these enriched cultures showed resistance to toxic concentrations of Dox and increased efficiency of mammosphere formation. In purified GFP + or GFP- cells, Dox increased the mammosphere-forming efficiency, promoted phenotypic switches in non-CSCs populations to a CSC-like state, reduced proliferation, and induced differential gene expression. We identified several biological processes and molecular functions that partially explain the development of doxorubicin-resistant cells and cellular plasticity. Among the genes that were regulated by Dox exposure, the expression of ITGB1, SNAI1, NOTCH4, STAT5B, RAPGEF3, LAMA2, and GNAI1 was significantly associated with poor survival, the stemness phenotype, and chemoresistance.The generation of chemoresistant cells that have characteristics of CSCs, after exposure to low concentrations of Dox, involves the differential expression of genes that have a clinical impact.© 2024. The Author(s).