Department of Breast Cancer Surgery, Harbin Medical University Cancer Hospital, Harbin Medical University, Haping RD NO, 150086, Harbin, Heilongjiang Province, People's Republic of China.
North China Translational Medicine Research Center of Harbin Medical University, Harbin Medical University, Harbin, 150086, Heilongjiang, China.
Naunyn Schmiedebergs Arch Pharmacol. 2023 Oct;396(10):2545-2553. doi: 10.1007/s00210-023-02477-7. Epub 2023 Apr 24.
Breast cancer stem cells (BCSCs) have been suggested to contribute to chemotherapeutic resistance and disease relapse in breast cancer. Thus, BCSCs represent a promising target in developing novel breast cancer treatment strategies. Mitochondrial dynamics in BCSCs were recently highlighted as an available approach for targeting BCSCs. In this study, a three-dimensional (3D) cultured breast cancer stem cell spheres model was constructed. Mitochondrial dynamics and functions were analyzed by flow cytometry and confocal microscopy. We have demonstrated that the protein levels of FIS 1 and Mitofusin 1 were significantly increased in BCSCs. Moreover, Capivasertib (AZD5363) administration could suppress Mitofusin1 expression in BCSCs. Our use of MitoTracker Orange and annexin V double-staining assay suggested that AZD5363 could induce apoptosis in BCSCs. The sensitivity of stem cell spheres to doxorubicin was investigated by CCK8 assay, and our results indicated that AZD5363 could re-sensitize BCSCs to Doxo. Flow cytometry analysis identified doxo-induced CD44 and CD133 expression in BCSCs could be suppressed by AZD5363. In combination with AZD536, doxo-induced apoptosis in the BCSCs was significantly increased. In conclusion, our study explored, for the first time, that AZD5363 could target mitochondrial dynamics in 3D cultured stem cell spheres (BCSCs) by regulating Mitofusin.
乳腺癌干细胞（BCSCs）被认为有助于乳腺癌的化疗耐药和疾病复发。因此，BCSCs 是开发新型乳腺癌治疗策略的有前途的靶点。最近，BCSCs 中的线粒体动力学被强调为靶向 BCSCs 的一种可行方法。在本研究中，构建了三维（3D）培养乳腺癌干细胞球体模型。通过流式细胞术和共聚焦显微镜分析线粒体动力学和功能。我们已经证明 FIS1 和 Mitofusin1 的蛋白水平在 BCSCs 中显著增加。此外，Capivasertib（AZD5363）给药可以抑制 BCSCs 中的 Mitofusin1 表达。我们使用 MitoTracker Orange 和 annexin V 双重染色测定法表明，AZD5363 可以诱导 BCSCs 凋亡。通过 CCK8 测定法研究了干细胞球体对多柔比星的敏感性，结果表明 AZD5363 可以使 BCSCs 重新对 Doxo 敏感。流式细胞术分析鉴定出 AZD5363 可以抑制 Doxo 诱导的 BCSCs 中 CD44 和 CD133 的表达。与 AZD536 联合使用时，Doxo 诱导的 BCSCs 凋亡明显增加。总之，我们的研究首次探索了 AZD5363 通过调节 Mitofusin 靶向 3D 培养干细胞球体（BCSCs）中的线粒体动力学。
