Apoptosis induction in MOLT-4 Cell line using TRAIL and SAHA and study of intracellular resistance mechanisms to TRAIL

Abstract

T-cell acute lymphoblastic leukemia is a lymphoid tumor with a fast progression rate and high complications. TRAIL can be used as a new strategy to induce apoptosis in cancer cells without harming normal healthy cells, but almost half of cancer cells show resistance to this agent through various mechanisms. Histone deacetylase inhibitors such as SAHA can be used to increase the sensitivity of cancer cells to TRAIL-induced apoptosis. In this study, we tested this hypothesis on MOLT-4 cancer cells. Materials and methods: First, IC50 for SAHA was determined using the MTT method. Then the cells were divided into six groups including the control group, TRAIL 50 nM, TRAIL 100 nM, SAHA 2 µM, SAHA 2 µM + TRAIL 50 nM, and SAHA 2 µM + TRAIL 100 nM. Then the amount of apoptosis was measured by flow cytometry using Annexin-V staining and PI after 24, 48 and 72 hours. The expression levels of c-flip, DR4, DR5, CHOP, NF-κB, STAT3, Akt, and PI3K genes were evaluated by quantitative real-time PCR method. Data were analyzed using two-way variance analysis with Tukey's and Dunnett's multiple comparisons tests, and p values less than 0.05 were considered statistically significant. Results: It was observed that the combination of SAHA with TRAIL increases apoptosis in MOLT-4 cells by increasing the expression of DR4, DR5, and CHOP and decreasing the expression of c-flip, STAT3, PI3k, Akt, and NF-kB genes.