Novel Combination of Arsenic Trioxide (As₂O₃) Plus Resveratrol in Inducing Programmed Cell Death of Human Neuroblastoma SK-N-SH Cells

Abstract

Aim: Arsenic trioxide (As 2 O 3), known as pi-shuang and the most toxic compound in traditional Chinese medicine, has been used as an antitumor agent for thousands of years. Resveratrol (3,5,4'-trihydroxy-trans-stilbene) is a natural phenol that has significant anti-bacterial, anti-fungaI and antiaging activities. Our study aimed to examine the combined anticancer effects of As 2 O 3 and resveratrol against human neuroblastoma SK-N-SH cells, and elucidate the underlying intracellular signaling. Materials and Methods: SK-N-SH cells were treated with an extremely low-dose (2-4 μM) of As 2 O 3 alone or combined with 75 μg/ml resveratrol for further comparisons. Cell viability, apoptotic signaling as well as synergistic cytotoxic effects were estimated using the MTT assay, microscopy observation, flow cytometric analysis for loss of mitochondrial membrane potential (MMP) and reactive oxygen species (ROS), and typical quantitative western blotting analysis. Student's ttest, and one-and two-way analysis of variance (ANOVA) were used for examination of significant differences. Results: The combined treatment was more effective than single treatment of As 2 O 3 or resveratrol alone in suppressing cell viability, which correlated with the elevation of ROS levels. The intracellular mechanisms of cytotoxicity of As 2 O 3 plus resveratrol were revealed as ROS accumulation and relative decrease of MMP, leading to activation of caspase-3 and-9, but not of caspase-1,-7 and-8. Combination treatment reduced the expression of B-cell lymphoma 2 (BCL2), BH3 interacting domain death agonist (BID), and BCL-x/L. Conclusion: Combined treatment at extremely low concentration of two agents from natural products, As 2 O 3 and resveratrol, has high potential as a cocktail of anticancer drugs for neuroblastoma. Neuroblastoma is listed as the most fatal cancer among children worldwide, and exhibits a very complex biological and tremendous clinical heterogeneity (1). Remarkably, neuroblastoma is renowned for having no obvious environmental or genetic risk factors, no effective drugs, and low 5-year survival, with parents and relatives of the pediatric patient consequently suffering from much emotional and psychological pain. Clinically, the age and gender of the patient, and the stage and molecular defects are key determinants for the efficacy of treatments, prognosis and modalities of therapy. At the time of diagnosis, about 453 This article is freely accessible online.

Novel 3‐aryl‐5‐substituted‐coumarin analogues: Synthesis and bioactivity profile

“…SK‐N‐SH human neuroblastoma cell line was selected to allow comparisons on the cytotoxic activity evaluation of hydroxy and oxyprenylated coumarin derivatives studied previously by our group (Kavetsou et al, ). Both SK‐N‐SH and HeLa cells have been widely used to examine the anticancer effect of bioactive agents (Akram et al, ; Dickens et al, ; Goswami et al, ; Park et al, ; Qi et al, ; Swift, Zhang, Trippett, & Narendran, ; Yen et al, ).…”

Resveratrol modulates the apoptosis and autophagic death of human lung adenocarcinoma A549 cells via a p53‑dependent pathway: Integrated bioinformatics analysis and experimental validation

“…More recently, autophagy, a catabolic process involving the degradation of unwanted macromolecules and organelles, has gained increasing attention, and is considered a double-edged sword with regard to tumorigenesis, development and therapy; autophagy may be used as a survival mechanism induced during adverse conditions, or conversely, a mode of cell death known as autophagic cell death, also termed type II PCD ( 13 ). Numerous studies have suggested that RSV may exert its anticancer effects via the induction of cell apoptosis individually or jointly in various types of cancer ( 14 - 16 ). Furthermore, studies have also suggested that RSV may initiate cell autophagy accompanied with or without apoptosis ( 17 , 18 ).…”

Arsenic Trioxide Increases Apoptosis of SK-N-BE (2) Cells Partially by Inducing GPX4-mediated Ferroptosis

“…Previous studies have shown that ATO inhibited NB cells growth via activation of apoptosisrelated proteins [29]. Lowest dose of ATO could induce the oxidative stress and cause the alterations of BCL2, BID and BCL-x/L, and activation of caspase-3 and-9, resulting in activation of apoptotic signaling pathways [30]. However, the detailed molecular mechanisms of ATO action are is still largely unknown.…”