Assessing Autophagy Flux in Glioblastoma Temozolomide Resistant Cells.

Q4 Biochemistry, Genetics and Molecular Biology Methods in molecular biology Pub Date : 2024-09-28 DOI:10.1007/7651_2024_571
Courtney Clark, Amir Barzegar Behrooz, Marco Cordani, Shahla Shojaei, Saeid Ghavami
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Abstract

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.

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评估胶质母细胞瘤替莫唑胺耐药细胞的自噬通量
自噬是参与细胞质成分降解和再循环的关键细胞过程,在癌症中发挥着促进细胞存活或促进细胞死亡的双重作用。在胶质母细胞瘤(GB)中,自噬与化疗药物替莫唑胺(TMZ)的抗药性有关。本研究提出了一种新方法,将先进的成像技术与生化检测相结合,精确测量对替莫唑胺(TMZ)耐药的胶质母细胞瘤细胞中的自噬通量。通过量化 LC3-II 和 SQSTM1 等关键自噬标记物,我们的方法提供了在治疗压力下自噬体形成和清除动态过程的详细见解。这种方法加深了我们对国标化疗耐药性中自噬作用的理解,对开发自噬靶向疗法具有重要意义。实时监测和操纵自噬通量的能力为监测和了解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种前景广阔的途径。
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来源期刊
Methods in molecular biology
Methods in molecular biology Biochemistry, Genetics and Molecular Biology-Genetics
CiteScore
2.00
自引率
0.00%
发文量
3536
期刊介绍: For over 20 years, biological scientists have come to rely on the research protocols and methodologies in the critically acclaimed Methods in Molecular Biology series. The series was the first to introduce the step-by-step protocols approach that has become the standard in all biomedical protocol publishing. Each protocol is provided in readily-reproducible step-by-step fashion, opening with an introductory overview, a list of the materials and reagents needed to complete the experiment, and followed by a detailed procedure that is supported with a helpful notes section offering tips and tricks of the trade as well as troubleshooting advice.
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