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Updated: Jun 10, 2025
Pre-clinical Evaluation of Tyrosine Kinase Inhibitors for Treatment of Acute Leukemia
Published on: September 18, 2013
Study of TRAIL and SAHA Co-Treatment on Leukemia K562 Cell Line
Amirarsalan Alaei1,2, Saeed Solali3, Masoud Mohammad Mirzapour1
1Immunology Research Center, Tabriz University of Medical Sciences, Tabriz, Iran.
Abstract:
TRAIL (Tumor Necrosis Factor-Related Apoptosis-Inducing Ligand) is an attractive agent being considered a potential cancer treatment. It attaches to its death receptors, leading many cancer cells to apoptosis. However, some malignancies indicate substantial resistance to TRAIL, challenging 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 the leukemia K562 cell line. First, the IC50 for SAHA was calculated (2 µM) at 12, 24, 48, and 72 h of treatment using MTT assay. 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 h and the survival of these cells was evaluated by Flowcytometry following the annexin-V and PI staining. To demonstrate the non-toxicity of the combined treatment for normal cells, the HEK-293 cell line was treated with the TRAIL 100 nM and SAHA 2 μM combined and separated at the same periods. In the end, by performing real-time PCR, the amount of candidate genes' expression implicated in TRAIL resistance, and the levels of BCR-ABL expression was measured. The drug dosages were not toxic to normal cells. SAHA plus TRAIL strongly triggered apoptosis in K562 cells after 24, 48, and 72 h of exposure. Furthermore, it was shown that DR4, DR5, and CHOP expressions were enhanced, and PI3K, Akt, ERK, STAT3, c-FLIPL, NF-κB, and BCR-ABL expressions were decreased by SAHA in K562 cells. Our study indicated that SAHA combined with TRAIL can increase the sensitivity of K562 leukemic cells to TRAIL by suppressing intracellular anti-apoptotic molecules and augmenting the expressions of DR4/DR5 and CHOP.
Insights
Suberoyl anilide hydroxamic acid (SAHA) combined with Tumor Necrosis Factor-Related Apoptosis-Inducing Ligand (TRAIL) effectively triggers apoptosis in K562 leukemia cells, overcoming TRAIL resistance by modulating gene expression. This combination therapy shows promise for leukemia treatment.
Area of Science:
- Oncology
- Molecular Biology
- Pharmacology
Background:
- Tumor Necrosis Factor-Related Apoptosis-Inducing Ligand (TRAIL) is a promising anti-cancer agent that induces apoptosis in cancer cells.
- TRAIL resistance is a significant challenge in treating certain malignancies, including leukemia.
- Suberoyl Anilide Hydroxamic Acid (SAHA) is a histone deacetylase inhibitor with potential anti-cancer properties.
Purpose of the Study:
- To investigate the efficacy of combining SAHA with TRAIL in overcoming TRAIL resistance in K562 leukemia cells.
- To evaluate the impact of SAHA and TRAIL combination therapy on apoptosis induction and gene expression in leukemia cells.
- To assess the safety of the combined treatment on normal human cells.
Main Methods:
- K562 and HEK-293 cell lines were treated with varying concentrations of TRAIL and SAHA.
- Cell viability and apoptosis were assessed using MTT assays and flow cytometry (annexin-V/PI staining).
- Gene expression levels of apoptosis-related factors (DR4, DR5, CHOP) and signaling pathways (PI3K, Akt, ERK, STAT3, c-FLIPL, NF-κB, BCR-ABL) were analyzed using real-time PCR.
Main Results:
- SAHA demonstrated an IC50 of 2 µM in K562 cells.
- The combination of SAHA and TRAIL significantly induced apoptosis in K562 cells at 24, 48, and 72 hours.
- The combined treatment upregulated DR4, DR5, and CHOP expression while downregulating PI3K, Akt, ERK, STAT3, c-FLIPL, NF-κB, and BCR-ABL in K562 cells.
- Drug dosages were not toxic to normal HEK-293 cells.
Conclusions:
- SAHA effectively overcomes TRAIL resistance in K562 leukemia cells when used in combination with TRAIL.
- The combination therapy enhances apoptosis by suppressing anti-apoptotic molecules and upregulating pro-apoptotic factors.
- This combination strategy holds potential for the treatment of TRAIL-resistant leukemia.
