Role Of G1P3 In Inducing Mitochondrial Derived Vesicle and Its Link to Endomembrane Vesicle
Document Type
Thesis
Degree Name
Master of Science (MS)
Department
Biological and Environmental Sciences
Date of Award
8-14-2024
Abstract
In the United States, breast cancer is the second most prevalent type of cancer among women, with 6% of those women developing metastasis. Previously reported, G1P3/IFI6 is an interferon (IFN)-stimulated gene with an anti-apoptotic property that elevates mitochondrial reactive oxygen species (mtROS) and promotes metastasis of G1P3-overexpressed breast cancer cells (MCF7/G1P3). Based on this, we hypothesized that G1P3 mediates the lysosomal degradation of ROS damaged mitochondria as a form of quality control. To test this hypothesis, G1p3 was down regulated using siRNA specific to G1P3 in MCF7/G1P3 cells, investigating mitochondria membrane potential and co-occurrence of mitochondrial outer marker PDH and inner marker TOMM20 with lysosomal marker LAMP1. Additionally, MCF7/VECTOR and MCF7/G1P3 immuno-stained cells were treated with ROS scavenger Mito TEMPO, mitochondria electron transport chain complex III inhibitor Antimycin A and lysosomal acidification inhibitor Bafilomycin A1to inspect the co-occurrence of TOMM20 with LAMP1 and PDH with LAMP1. Lastly, isolated mitochondria from MCF7/VECTOR and MCF7/G1P3 iv cells were resuspended in energy regenerating buffer to investigate mitochondrial vesical formation. Downregulating G1P3 expression in MCF7/G1P3 cells lowered mitochondrial membrane potential. Whereas in immunostaining result, knockdown of G1P3 suggests that in G1P3 cells, mitochondria region devoid of TOMM20 co-occur with lysosome which includes mitochondria matrix PDH. Additionally, Mito TEMPO result suggests reduced level of mtROS promotes the co-occurrence of TOMM20 free region of mitochondria with lysosome in MCF7/G1P3 cells. While Antimycin A result suggests that higher mtROS level decreases co-occurrence of TOMM20 with LAMP1 in G1P3 expressing cells unaffecting matrix association. In the same way, Bafilomycin A1 treatment result has similar effect as Antimycin A. Lastly, neither MCF7/VECTOR or MCF7/G1P3 cell’s resuspended mitochondria formed vesicles when in energy regenerating buffer. In summary, G1P3 promotes the co-occurrence of Mitochondrial region free of TOMM20 with lysosomes and is regulated by mtROS. Understanding the molecular route of G1P3 controlling mitochondrial cargo with respect to lysosomes may result in new approaches to the treatment of breast cancer.
Advisor
Venu gopalan Cheriyath
Subject Categories
Biology | Life Sciences
Recommended Citation
Shakya, Diksha, "Role Of G1P3 In Inducing Mitochondrial Derived Vesicle and Its Link to Endomembrane Vesicle" (2024). Electronic Theses & Dissertations. 1219.
https://digitalcommons.tamuc.edu/etd/1219
