Akademik Veri Yönetim Sistemi
Tez Türü: Doktora
Tezin Yürütüldüğü Kurum: The University of Manchester, Cancer Research, İngiltere
Tez Danışmanı: Kaye Williams, Adam Hurlstone
Tezin Onay Tarihi: 2025
Tezin Dili: İngilizce
Desteklendiği Program: Diğer
Özet:
Background: Endometrial cancer (EC) cells exhibit metabolic adaptability to survive in low-oxygen and nutrient-limited environments, utilizing resources like lactate, glucose and glutamine. This adaptability is essential for cancer cell survival, enabling tumour cells to thrive in challenging conditions. MCT1, a transporter for lactate and pyruvate, supports energy maintenance and growth by supporting metabolic reprogramming of cancer cells. MCT1 has been shown to have prognostic significance in various cancer types. Further, its subcellular localisation is shown to have an impact on overall survival in EC and soft-tissue sarcomas. Therefore, understanding the factors influencing MCT1 function and expression in EC is vital for exploiting this for patient benefit. This project aims to understand the effect of oxygen and nutrient availability on MCT1 expression and function, and expand the knowledge on the effect of nuclear MCT1 in EC.
Methods: Ishikawa (type I), HEC-1-A (type II), and AN3-CA (metastatic type I) cell lines, were used as models to advance knowledge on the role of MCT1 in EC. For this, various methods were used to evaluate expression (western blot, IF), metabolic activity (Seahorse, lactate and ROS assay), and functions (SRB, MTT, cell cycle and Annexin V) of the cells following siRNA KD or AZD3965 inhibition of MCT1 under different oxygen (21%, 3% and 1%) and nutrient-limited environments (L-Glutamine 0 mM and 2 mM).
Results: MCT1 knockdown led to a compensatory upregulation of MCT4, indicating a strong adaptive response to maintain lactate transport. Metabolic profiling revealed that the absence of L-glutamine significantly reduced mitochondrial respiration in Ishikawa cells, as demonstrated by decreased OCR. Functional analyses, including cell cycle assessment, annexin-V staining, and ROS measurements, showed distinct profiles resulting from MCT1 knockdown across the three cell lines and cells exhibited different responses to low-oxygen. In addition to these functional analyses, similar to EC human biopsies, nuclear localisation of MCT1was observed in all three cell lines and was detected in the chromosome structure.
Conclusions: The study offers important insights into the metabolic adaptations of EC cells in response to MCT1 inhibition/knockdown, hypoxic stress and glutamine-limited environments. The findings underscore the potential of MCT1 as a therapeutic target to disrupt the metabolic flexibility of cancer cells, thereby impairing their survival and metastatic potential. Furthermore, the presence of MCT1 in the nucleus suggests a non-canonical function for this transporter.