【目的】 建立人源 Luminal型乳腺癌干细胞模型并寻找有效干预乳腺癌干细胞生长的方法。【方法】 使用干细胞专用培 养基将MCF7乳腺癌细胞诱导成乳腺癌干细胞，接着将MCF7乳腺癌干细胞分为空白对照组及黄芪甲苷组，给予磷酸盐缓冲 液（PBS）或黄芪甲苷处理。观察药物干预后细胞形态变化，通过细胞 3D 球囊、细胞活力检测实验等明确黄芪甲苷的药效， 并分析上述处理后细胞转录组及相关基因的表达变化。【结果】 与MCF7乳腺癌细胞相比，MCF7乳腺癌干细胞球囊的生长速 度较快（P＜0.01），干细胞标志物醛脱氢酶 1家族成员 A1（ALDHA1）的表达显著增加（P＜0.01）。进一步与空白对照组相比， 黄芪甲苷组MCF7乳腺癌干细胞增殖减少（P＜0.001），细胞球囊的生长速度较缓慢（P＜0.01）。转录组分析显示，干细胞造模 及黄芪甲苷干预后，差异基因富集于细胞衰老信号通路。黄芪甲苷干预后 β-半乳化糖苷酶（SA-β-gal）阳性细胞数目较多 （P＜0.01）。RT-PCR 检测发现黄芪甲苷干预 MCF7 乳腺癌干细胞后，白细胞介素（IL）-1α（P＜0.01）、P21（P＜0.001）、P53 （P＜0.05）mRNA表达水平显著升高。【结论】 黄芪甲苷通过诱导细胞衰老抑制人源Luminal型乳腺癌干细胞的生长。

Objective To establish a human luminal breast cancer stem cell （BCSC） model and investigate the inhibitory effects of astragaloside IV （AS-IV） on BCSC growth. Methods MCF-7 breast cancer cells were cultured in stem cell-specific medium to induce BCSC formation. The BCSCs were then divided into a blank control group and an AS-IV treatment group， both groups were given PBS or AS-IV treatment. Morphological changes were observed after intervention. The therapeutic efficacy of AS-IV was evaluated using 3D spheroid formation and cell viability assays. Transcriptomic profiling and gene expression analysis were performed to elucidate the underlying mechanisms. Results Compared with the MCF7 breast cancer cells，MCF7 breast cancer stem cell mammospheres exhibited accelerated growth （P＜0.01） and significantly increased expression of the stemness marker ALDH1A1 （P＜0.01）. Further comparison with the blank control group revealed that astragaloside IV （AS-IV） treatment significantly inhibited MCF7 breast cancer stem cell proliferation （P＜0.001） and slowed mammosphere growth （P＜0.01）. Transcriptomic analysis demonstrated that differentially expressed genes （DEGs） induced by stem cell modeling and AS-IV intervention were enriched in the cellular senescence signaling pathway. AS-IV intervention substantially increased the number of SA-β-gal-positive cells （P＜0.01）. RT-PCR analysis confirmed that AS-IV significantly upregulated mRNA expression of IL-1α（P＜0.01），P21（P＜0.001），and P53（P＜0.05） in MCF7 breast cancer stem cells. Conclusion Astragaloside IV suppresses the growth of human luminal breast cancer stem cells by inducing cellular senescence.

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国家自然科学基金资助项目（编号：81974571、82274513、82474504）；广东省自然科学基金资助项目（编号：2020A15110760、 2023A1515011115）；中医药广东省实验室（横琴实验室）科技研发培植项目（编号：HQL2024PZ023）；广州市科技计划项目（编号： SL2023A03J01120、SL2023A04J00228）；广东省中医院中医药科技专项科研项目（编号：YN2022QN32）；广东省中医证候临床研究重点实验室 项目（编号：YN2023ZH10）；中医湿证国家重点实验室自主项目（编号：QZ2023ZZ13）；北京迈迪科公益基金会项目（编号：MDK2023-1007）； 北 京科创医学发展基金会项目（编号：KC2023-JX-0082-12、KC2021-ZZ-0010-9）；广东省中医药局科研项目（编号：20242035、 20251142）