PURPOSE: The treatment of ovarian cancer (OC) with chemotherapy leaves resistant cancer cells which in a short time re-grow as recurrent cancer. A diverse array of resistance mechanisms for chemotherapy has been described but none have proven as viable targets in a clinical setting. Cancer stem cells (CSCs) are increasingly accepted as the putative mediators of chemoresistance and relapse of cancer. This study aimed to understand the molecular mechanisms involved with chemoresistance and recurrence by investigating the proteomic profile of ascites-derived tumor cells obtained from OC patients prior to (chemonaive, CN) and after chemotherapy treatments (recurrent, CR).
METHODS: Ascites collected from CN and CR OC patients diagnosed with advanced-stage serous OC were cultured using a novel in vitro method to obtain a distinct population of epithelial tumor cells. Flow cytometry and immunofluorescence were used to characterize the tumor population. High-resolution label-free quantitative proteomic profiling was used to define significantly differentially expressed proteins between CN and CR tumor cells. KEGG and DAVID software's were used to determine pathways associated with CR cells. The mechanisms of survival of cisplatin and paclitaxel-treated OC cancer cells were determined in vitro by mRNA analysis and in vivo mouse xenograft models by immunohistochemistry.
RESULTS: Proteomic profiling of CN and CR tumor cells showed significant differences in proteins encoding for CSC, immune surveillance, DNA repair mechanisms, cytoskeleton rearrangement, cell-cell adhesion, cell cycle pathways, cellular transport, and proteins involved with glycine/proline/arginine synthesis in tumor cells isolated from CR relative to CN patients. Pathway analyses revealed enrichment of chemoresistance markers, energy metabolism, DNA repair mechanisms and immune surveillance pathways in recurrent CR tumor cells. Validation of these pathways in OC cell lines and nude mice treated with chemotherapy demonstrated increased expression of CSC and chemoresistant markers, enhancement in glycolytic pathway and oxidative phosphorylation, suppression of immune surveillance in chemotherapy treated cell lines and recurrent mice xenografts compared to untreated control.
CONCLUSION: These findings unravel some of the molecular mechanisms by which chemoresistance and relapse occur in OC patients' post-chemotherapy treatment; and may be important in designing novel therapeutic options for advanced-stage OC patients.
Citation Format: Nuzhat Ahmed, Ruth Escalona, Elif Kadife, Dilys Leung and George Kannourakis. CANCER STEM CELLS: DISTINCT SEEDS FOR RECURRENT OVARIAN CANCER [abstract]. In: Proceedings of the 12th Biennial Ovarian Cancer Research Symposium; Sep 13-15, 2018; Seattle, WA. Philadelphia (PA): AACR; Clin Cancer Res 2019;25(22 Suppl):Abstract nr AP26.