Understanding the Epithelial-Mesenchymal Transition (EMT) Leading to Metastasis in Early and Late-stage Ovarian Cancer Models

Mondal, Tanusree (2025) Understanding the Epithelial-Mesenchymal Transition (EMT) Leading to Metastasis in Early and Late-stage Ovarian Cancer Models. Masters thesis, Indian Institute of Science Education and Research Kolkata.

Text (MS Dissertation of Tanusree Mondal (20MS228)) 20MS228_Thesis_file.pdf - Submitted Version Restricted to Repository staff only Download (6MB)

Abstract

Ovarian cancer is among the deadliest gynaecological malignancies due to its high metastatic potential and resistance to therapy. While Vascular Endothelial Growth Factor (VEGF) is known as a key regulator of angiogenesis [25] in tumor progression, its roles in epithelial-mesenchymal transition (EMT), cellular stress response pathways, and cancer dormancy remain incompletely understood. For my MS thesis, I focused on the cellular and molecular aspects of cancer progression. My research aims to identify novel markers and pathways through which VEGF regulates EMT and metastasis, modulates various stress response pathways, and influences the induction and termination of cancer dormancy to understand cancer progression mechanisms better. I am working with ID8 and ID8-VEGF murine ovarian cancer cell lines. ID8 is a secondary cell line established from late-passaged ovarian surface epithelial cells of C57BL/6 mice, which shows slow tumor proliferation. In contrast, the VEGF-overexpressing ID8 strain (ID8-VEGF) was generated by transfection with a retroviral vector containing green fluorescent protein (GFP) and VEGFA, representing more aggressive ovarian cancer variants. In my experiments, cell culture, protein extraction, and western blotting were performed to assess the expression of key EMT markers (E-cadherin, Vimentin), heat shock proteins (HSF1, HSP70, HSP90), and other regulatory proteins (p53, p65). Immunofluorescence assays were conducted to examine the localization and expression of Occludin, Vimentin, β-catenin, and ZO-1. Additionally, real-time PCR was performed to analyze the mRNA expression levels of N-cadherin, NF-κB1, matrix metalloproteinases (MMP-3, MMP-9, MMP-12), and their inhibitors (TIMP-1, TIMP-2, TIMP-3). Cell migration ability was further evaluated using a scratch assay, where ID8-VEGF cells exhibited faster wound closure than ID8 cells, indicating enhanced migratory potential.

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