Clark Courtney, Barzegar Behrooz Amir, Cordani Marco, Shojaei Shahla, Ghavami Saeid
Department of Human Anatomy and Cell Science, Max Rady College of Medicine, Rady Faculty of Health Sciences, University of Manitoba, Winnipeg, MB, Canada.
Department of Biochemistry and Molecular Biology, Faculty of Biology, Complutense University of Madrid, Madrid, Spain.
Methods Mol Biol. 2025;2879:225-238. doi: 10.1007/7651_2024_571.
Autophagy is a critical cellular process involved in the degradation and recycling of cytoplasmic components, playing a dual role in cancer by either promoting cell survival or facilitating cell death. In glioblastoma (GB), autophagy has been implicated in resistance to the chemotherapeutic agent temozolomide (TMZ). This study presents a novel method to accurately measure autophagy flux in TMZ-resistant glioblastoma cells, combining advanced imaging techniques with biochemical assays. By quantifying key autophagy markers such as LC3-II and SQSTM1, our approach provides detailed insights into the dynamic processes of autophagosome formation and clearance under therapeutic stress. This method advances our understanding of autophagy in GB chemoresistance and has significant implications for the development of autophagy-targeted therapies. The ability to monitor and manipulate autophagy flux in real time offers a promising avenue for monitoring and understanding TMZ resistance and improving patient outcomes in glioblastoma treatment.
自噬是一种关键的细胞过程,参与细胞质成分的降解和再循环,在癌症中发挥双重作用,既促进细胞存活,也促进细胞死亡。在胶质母细胞瘤(GB)中,自噬与对化疗药物替莫唑胺(TMZ)的耐药性有关。本研究提出了一种新方法,将先进的成像技术与生化分析相结合,以准确测量TMZ耐药性胶质母细胞瘤细胞中的自噬通量。通过量化关键的自噬标志物,如LC3-II和SQSTM1,我们的方法提供了对治疗压力下自噬体形成和清除动态过程的详细见解。该方法增进了我们对GB化疗耐药性中自噬的理解,对自噬靶向治疗的发展具有重要意义。实时监测和操纵自噬通量的能力为监测和理解TMZ耐药性以及改善胶质母细胞瘤治疗中的患者预后提供了一条有前景的途径。