Investigating autophagy and mitophagy in 12Z endometriosis cells via exposure to pharmacological inhibitors

Investigating autophagy and mitophagy in 12Z endometriosis cells via exposure to pharmacological inhibitors DOI: https://doi.org/10.13021/jssr2025.5187Abstract Endometriosis is a chronic condition in which endometrial tissue spreads outside the uterus and causes inflammation. As these cells enter oxidative stress conditions, they rely on autophagy and mitophagy–cellular recycling mechanisms–to survive. Thwarting these processes may prohibit further growth. This experiment applied medications suspected to inhibit autophagy and mitophagy to endometriosis cells to study the effects on growth and protein pathways. Endometriosis 12Z cells were cultured and divided into two assays. In the first, cells were treated with chloroquine phosphate, metformin, lidocaine, or combinations of these for different time stamps. After treatment, cells were lysed and analyzed using reverse phase protein array (RPPA). In the second experiment, endometriosis cells were cultured and scratched to assess migration and growth response to metformin, lidocaine, tamoxifen, and chloroquine phosphate. Following treatment, cells were lysed and analyzed using western blot, RPPA, and immunohistochemistry. Morphological analysis of 12Z cells in the first experiment revealed responses to chloroquine phosphate, including rod-shaped distortion, refractility, and large cytoplasmic vacuoles. These effects were amplified by co-treatment with lidocaine or metformin, leading to pronounced cell swelling and vacuolization—signs of disrupted autophagy. Preliminary data suggests that 12Z cells exposed to drugs, such as chloroquine phosphate, left wider gaps in scratch assays, suggesting slower growth and increased stress responses. Metformin+Lidocaine increased PHGDH, Calbindin, and AMPK alpha Thr172 in the 48-hour time course. Chloroquine phosphate increased proteins in glucose, autophagy, and oxidative stress pathways. Chloroquine-treated cells induced autophagy-disruption and stress, while Metformin+Lidocaine elevated the levels of autophagy pathway proteins. Ongoing research aims to investigate the pathways involved. Published Issue Section License This work is licensed under a Creative Commons Attribution-ShareAlike 4.0 International License.