细胞质体介导的自噬（CMA）通路在不同类型的癌症中失控，然而，其在癌症干细胞（CSCs）中的作用尚不明确。卵巢癌的发展，是最致命的妇科癌症类型之一，涉及将CSCs转移到腹腔。该研究的目的是确定CMA在卵巢CSCs中的作用。我们发现，转录因子EB（TFEB）和海藻糖，一种诱导TFEB活化的二糖，增强了八聚体结合转录因子4（OCT4）干细胞和溶酶体相关膜蛋白2A（LAMP2A）CMA标记的表达。然而，海藻糖没有增加卵巢CSCs中LC3II的宏自噬标记水平。在A2780和SKOV3卵巢CSCs中，LAMP2A和热休克蛋白70（HSC70）的表达水平比正常粘附细胞高。我们的结果表明，LAMP2A基因的沉默导致卵巢CSCs中球形体形成减少，并增强了GLUT5的表达。此外，果糖处理减少了卵巢CSCs的球形体形成，并增强了LAMP2A、SOX2和OCT4的表达水平。KEGG功能分析显示，经过果糖处理的A2780-spheroid（SP）细胞中已不同表达基因富集在铁死亡通路中。在A2780-SP和SKOV3-SP细胞中，果糖处理后，SLC7A11的水平降低，而FTH的水平增加。综上所述，我们的结果表明，通过果糖代谢途径介导CMA在CSCs中的作用。版权所有 © 2023 Elsevier B.V. 发布。

The chaperone-mediated autophagy (CMA) pathway is deregulated in different types of cancers; however, its role in cancer stem cells (CSCs) is unknown yet. Development of ovarian cancer, the most lethal gynecological type of cancer, involves the metastasis of CSCs to the abdominal cavity. This study aims to determine the role of CMA in ovarian CSCs. We found that the transcription factor EB (TFEB) and trehalose, a disaccharide that induces TFEB activation, enhance the expression of octamer-binding transcription factor 4 (OCT4) stem cell and lysosomal-associated membrane protein 2A (LAMP2A) CMA markers. However, trehalose did not increase the level of the LC3II macroautophagy marker in ovarian CSCs. In A2780 and SKOV3 ovarian CSCs, LAMP2A and heat shock protein 70 (HSC70) exhibited higher expression levels than in normal adherent cells. Our results showed that the silencing of the LAMP2A gene resulted in reduced sphere formation and enhanced GLUT5 expression in ovarian CSCs. Moreover, the treatment with fructose reduced sphere formation and enhanced the expression levels of LAMP2A, SOX2, and OCT4 in ovarian CSCs. The KEGG functional analysis revealed that differentially expressed genes were enriched in the ferroptosis pathway in A2780-spheroid (SP) cells after treatment with fructose. In A2780-SP and SKOV3-SP cells, the level of SLC7A11 decreased whereas FTH increased after treatment with fructose. Taken together, our results suggest that CMA is mediated in CSCs via fructose metabolism.Copyright © 2023. Published by Elsevier B.V.