Inhibiting AKT signaling pathway with cilostazol and meloxicam synergism for suppressing K562 cells in vitro

Abstract

Despite advances in cancer treatment, chronic myeloid leukemia (CML) is still one of the leading causes of death in the world. Due to the role of inflammation in cancer promotion and progression, thus use of anti‐inflammatory agents may suppress cancer cell growth. In this study, we used two anti‐inflammatory drugs, cilostazol and meloxicam, for the treatment of CML. Cell viability was measured using 3‐(4,5‐dimethylthiazol‐2‐yl)‐2,5‐diphenyltetrazolium bromide (MTT) assay and the synergism occurrence was calculated by compusyn software. Annexin V/PI test and Hoechst staining were used to determine the apoptosis rate. To determine the pathway of apoptosis induction, the expression of BCL2 Associated X (Bax) and B‐cell lymphoma‐2 (Bcl‐2) apoptotic genes and caspases activity were evaluated. The cell cycle was analyzed by propidium iodide (PI) staining and flow cytometry. Western blot analysis and immunofluorescence were performed to estimate alterations in Ak strain transforming‐1 (AKT‐1), phosphprylated AKT‐1 (p‐AKT‐1), adenosine mono‐phosphate‐kinase (AMPK), and phosphorylated AMPK (p‐AMPK) proteins and BCR/ABL and c‐Myc distribution, respectively. Results showed that cilostazol, meloxicam, and their combination drug reduced cell viability (p < 0.05). Compared with control, expression of Bax and Bcl‐2 decreased in treated cells, respectively (p < 0.05). The caspase‐9 activity increased in treated cells compared to control cells (p < 0.001). The applied drugs decreased the protein level of p‐AKT‐1 while increasing the p‐AMPK protein level (p < 0.05). BCR/ABL and c‐Myc Protein distribution significantly decreased in treated cells. In conclusion, the combination drug had more cytotoxic effects than cilostazol and meloxicam alone and induced apoptosis by inhibiting AKT‐1 activation and c‐Myc reduction. Therefore using combination drugs effectively can treat cancers of CML origin.

Design, synthesis, in vitro and in silico evaluation of indole-based tetrazole derivatives as putative anti-breast cancer agents

“…It has demonstrated potential therapeutic effects in various types of cancer through multiple mechanisms, such as apoptosis induction or modulating various signalling pathways implicated in cancer development and progression. 34,35 Also, sorafenib analogues containing a tetrazole moiety showed anti-proliferative activity superior to sorafenib against HT-29, HepG2, MCF-7, and A549 cells. 36 Recently, a tetrazole derivative N-(5-(4-fluorobenzyl)thiazol-2yl)-4-(1H-tetrazol-1-yl)benzamide has been reported to have cytotoxicity against the breast cancer cell line T-47D with an IC 50 value of 896 nM.…”