Apoptosis induction in K562 cell line using TRAIL and SAHA and investigating the genes associated with resistance to TRAIL

Abstract

Cancer therapy remains a significant concern for researchers, and Numerous studies have been conducted in this area. TRAIL (Tumor Necrosis Factor-Related Apoptosis-Inducing Ligand) is an attractive agent that attaches to its death receptors, leading many cancer cells to apoptosis. However, some malignancies indicate substantial resistance to TRAIL, presenting a challenge for anticancer scientists. Herein, combination therapy with TRAIL plus SAHA (Suberoyl Anilide Hydroxamic Acid) was conducted to evaluate the capability of SAHA to overcome TRAIL resistance in leukemia K562 cell line. Methods and materials: First, the MTT assay was used to calculate the IC50 for SAHA (2 µM) at 12, 24, 48, and 72 hours of treatment. Second, the K562 cells were treated with concentrations of 50 and 100 nM of TRAIL and 2 μM of SAHA separately and together for 24, 48, and 72 hours and the survival of these cells was evaluated by Flowcytometry following the annexin-V and PI staining. In the end, performing real-time PCR, the amount of candidate genes' expression implicated in TRAIL resistance was measured. Results: SAHA combined with TRAIL strongly triggered apoptosis in K562 cells after 24, 48, and 72 hours of exposure. Furthermore, it was shown that DR4, DR5, and CHOP expressions were enhanced, and PI3K, Akt, ERK, STAT3, c-FLIP, and NF-κB expressions were decreased by SAHA in K562 cells.