Assessing Autophagy Activation in Advanced Ovarian Cancer Using Ascitic Fluid: A Feasibility Study

Introduction:
Autophagy contributes to chemotherapy resistance by promoting cell survival under stress in various malignancies, including ovarian cancer. The analysis of autophagy biomarkers in ascitic fluid represents an emerging approach with potential advantages over tissue-based studies. This study aimed to establish standardized and reproducible laboratory methods for detecting and quantifying autophagy biomarkers in the ascitic fluid of ovarian cancer patients.
Methods:
Ascitic fluid samples from 30 ovarian cancer patients were analyzed using three techniques:
1. Enzyme-linked immunosorbent assay (ELISA): Measured levels of Beclin 1, p62/sequestosome 1 (p62/sqstm1), and synaptosomal-associated protein 23 (SNAP 23).
2. Immunocytochemistry (ICC): Assessed the localization of Syntaxin 17 and vesicle-associated membrane protein 8 (VAMP 8).
3. Flow cytometry: Identified epithelial tumor cells and evaluated Annexin V expression as a pro-apoptotic marker.
Results:
We successfully standardized methods for assessing autophagy marker expression in ascitic fluid. Despite the small sample size, preliminary findings indicated variations in biomarker expression across different Autophinib disease stages. Notably, Beclin 1 levels were higher in relapsed patients compared to newly diagnosed cases, suggesting increased autophagy activation. ICC revealed heterogeneous expression patterns of Syntaxin 17 and VAMP 8 among patients. Flow cytometry identified tumor epithelial cells and confirmed Annexin V expression in these cells.
Conclusion:
This study established reproducible techniques for analyzing autophagy markers in ascitic fluid, providing a non-invasive and accessible method for studying ovarian cancer biology. Further research with larger cohorts and integration of traditional biomarkers could enhance its clinical applicability and improve disease management.