Evaluating the effect of simultaneous K-Ras and Akt genes inhibition in changing the sensitivity to chemotherapy drugs in mutant and normal K-Ras colorectal cell lines

Abstract

Mutations in K-Ras genes drive about 30% of human cancers, causing continuous pathway activation. Targeting K-Ras alone often activates the AKT pathway. Thus, inhibiting both K-Ras and AKT shows promise, but current treatments lack effective dual-targeting strategies, requiring new approaches.Objectives:Our study aims to evaluate the efficacy of newly designed shRNAs in suppressing AKT and K-Ras protein expression in two cancer cell lines, CACO2 (wild type K-Ras) and AGS (K-Ras(G12D)). By doing so, we seek to determine the feasibility of these shRNAs as promising therapeutic targets.Methods:Plasmid vectors encoding the designed shRNAs were used to generate recombinant plasmids, which were then transfected into CACO2 (wild-type K-Ras) and AGS (K-Ras(G12D)) cancer cell lines. Cell viability was assessed using MTT assays, and apoptotic effects were measured via flow cytometry. The inhibitory impact of the shRNAs was confirmed through cobalt-induced treatment and RT-PCR analysis. Comparative analysis between wild-type and mutant K-Ras cell lines will be conducted to determine the outcomes. Results: The shRNAs reduced AKT and K-Ras protein expression in cancer cell lines, decreasing viability and inducing apoptosis. Cobalt treatment and RT-PCR confirmed these effects. Comparative analysis supported the approach’s efficacy.Conclusion: the designed shRNAs effectively targeted AKT and K-Ras in cancer cells, demonstrating promising potential as a therapeutic approach for colorectal cancer.