Abstract 2413: TP53 gene mutations differently regulate ovarian cancer metabolism: Ex vivo and in vitro studies

Abstract

Abstract Background: Even in the presence of oxygen, cancer cells tend to metabolize glucose into lactate instead of undergoing oxidative phosphorylation. Knowing that glucose is crucial for tumor's proliferation, new patient's treatment strategies tend to combine chemotherapy to hypoglycemic drugs. p53 is a tumor suppressor and key regulator of glycolysis in cancer cells, however highly mutated in tumors. In ovarian cancer, the majority of studies concerning p53 mutations focus on the DNA binding domain since the majority of hotspot mutations affects this region of the gene. However, mutations affecting other regions such as the proline rich domain linked directly to apoptosis may also affect the protein's expression and activity. The aim of this study is to investigate the effect of various positions of mutations in TP53 gene on glycolysis, apoptosis and transcription of p53 target genes. Material and Methods: this study was divided into two parts. The in vitro study was done using 3 ovarian cancer cell lines SKOV-3 (p53 null), Igrov-1 (WT p53) and Ovcar-3 (DNA domain mutant p53), along with the transfection of SKOV-3 cells by three different vectors: WTp53, P72R p53 and R249S p53. The ex vivo part was performed on 30 ovarian cancer biopsies. The biopsies were subjected to Tp53 immunohistochemistry and sequencing to study the mutations and their effect on p53 expression. The effect of Tp53 mutations on glucose metabolism was assayed by glucose consumption measurements, lactate production and mRNA expression of various glycolytic enzymes. To study the effect of mutations on the transcriptional activity of p53, p21 and MDM2 expression was quantified in these cells. Annexin V assay was performed on transfected SKOV-3 to study the apoptotic activity of WT and mutant p53. Results: DNA binding domain mutant cells (Ovcar-3 and R249S SKOV-3) presented a 30% increase in glucose consumption and lactate production when compared to IGROV-1 and WT SKOV-3. P72R SKOV-3 exhibited the same pattern as WTp53 cells. The increase in the mRNA expression of glucose transporters GLUT-1 and 3 and of the glycolytic enzymes PK, PFK, G6PDH and aldolase in cells with DNA binding domain mutations varied from 20 to 30%, whereas the levels of PDHa and TIGAR decreased significantly (40%). The transcriptional activity of DNA binding domain mutant cells showed a 50% decrease in comparison to WT and P72R mutant cells. The apoptotic activity of p53 was unaltered in P72R transfected SKOV-3 cells. The ex vivo results revealed a high frequency of the P72R variant in the ovarian biopsies (69%), associated with an overexpression of the p53 protein. The glycolytic and transcriptional patterns of cells extracted from these biopsies were similar to those obtained in transfected cells. Conclusion: Though both DNA-binding and proline-rich domains mutations lead to p53 overexpression, only the first type promotes Warburg effect and could have the worst patient's prognosis. Citation Format: Stephanie Antoun, David Atallah, Roula Tahtouh, Malak Moubarak, Nada Alaeddine, George Hilal. TP53 gene mutations differently regulate ovarian cancer metabolism: Ex vivo and in vitro studies [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr 2413.