Abstract 534: Sotorasib-resveratrol combination induces synergistic cytotoxicity by downregulating K-RAS signaling in K-RAS mutant lung cancer cell lines

Abstract Background: K-RAS mutations are the most frequent (35-50%) in patients with non-small cell lung cancer (NSCLC) who have a smoking history. These mutations promote cancer development through constitutive activation of poliferative signaling pathways, metabolic modifications and mechanisms of apoptosis evasion. Sotorasib is a covalent inhibitor targeting KRAS G12C; however, the primary challenge lies in acquired resistance. Resveratrol is a natural compound that promotes apoptosis and inhibits proliferation through the MAPK and Wnt/β-catenin pathways in cancer cells. Therefore, this project proposes using resveratrol as an adjuvant therapy to sotorasib in cell lines with KRAS mutations. Methods: Dose-response curves were generated using MTT assays to determine the IC50 for sotorasib and the IC10 for resveratrol, in addition to determine the type of interaction between both drugs in the K-RAS mutant cell lines A549 (G12S) and H23 (G12C). Crystal violet assays were performed to measure the effect on cell proliferation. Mitochondrial membrane potential was assessed via flow cytometry. Finally, protein expression, activation, and inactivation were analyzed using western blot assays (K-RAS, BRAF, C-RAF, p-CRAF, GLUT1, GLUT3, HXK 1, AMPK and p-AMPK). Results: Our results revealed a synergistic effect of the combined sotorasib-resveratrol treatment in the H23 cell line, while an additive or antagonic effect was observed in the A549 cell line. Cell proliferation assays demonstrated significant growth inhibition in both cell lines when treated with sotorasib alone or in combination with resveratrol. Additionally, mitochondrial matrix collapse was predominantly observed with the combined treatments. Western blot analysis showed strong inhibition of the K-RAS signaling pathway and glucose transporters (GLUTs), along with AMPK activation, particularly in treatments involving resveratrol. Furthermore, the induced cytotoxicity was linked to the inhibition of K-RAS signaling, GLUTs, Hexokinase 1, and AMPK reactivation. Conclusions: The addition of resveratrol to sotorasib enhanced growth inhibition, independent of KRAS mutations, primarily through apoptotic mechanisms and the downregulation of the glucose metabolic pathway. Citation Format: Pedro Barrios Bernal, Norma Hernandez-Pedro, José Lucio-Lozada, Eunice Romero-Nuñez, Cesar Castillo-Ruiz, Brandon Lopez, Sandra Hernandez-Sanchez, Oscar Arrieta. Sotorasib-resveratrol combination induces synergistic cytotoxicity by downregulating K-RAS signaling in K-RAS mutant lung cancer cell lines [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2025; Part 1 (Regular Abstracts); 2025 Apr 25-30; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2025;85(8_Suppl_1):Abstract nr 534.