Sensitivity of allyl isothiocyanate to induce apoptosis via ER stress and the mitochondrial pathway upon ROS production in colorectal adenocarcinoma cells

Abstract

Allyl isothiocyanate (AITC), a bioactive phytochemical compound that is a constituent of dietary cruciferous vegetables, possesses promising chemopreventive and anticancer effects. However, reports of AITC exerting antitumor effects on apoptosis induction of colorectal cancer (CRC) cells in vitro are not well elucidated. The present study focused on the functional mechanism of the endoplasmic reticulum (ER) stress-based apoptotic machinery induced by AITC in human colorectal cancer HT-29 cells. Our results indicated that AITC decreased cell growth and number, reduced viability, and facilitated morphological changes of apoptotic cell death. DNA analysis by flow cytometry showed G2/M phase arrest, and alterations in the modulated protein levels caused by AITC were detected via western blot analysis. AITC also triggered vital intrinsic apoptotic factors (caspase-9/caspase-3 activity), disrupted mitochondrial membrane potential, and stimulated mitochondrial-related apoptotic molecules (e.g., cytochrome c, apoptotic protease activating factor 1, apoptosis-inducing factor, and endonuclease G). Additionally, AITC prompted induced cytosolic Ca 2+ release and Ca 2+-dependent ER stress-related signals, such as calpain 1, activating transcription factor 6α, glucose-regulated proteins 78 and 94, growth arrest-and DNA damage-inducible protein 153 (GADD153), and caspase-4. The level of reactive oxygen species (ROS) production was found to induce the hallmark of ER stress GADD153, proapoptotic marker caspase-3, and calpain activity after AITC treatment. Our findings showed for the first time that AITC induced G2/M phase arrest and apoptotic death via ROS-based ER stress and the intrinsic pathway (mitochondrial-dependent) in HT-29 cells. Overall, AITC may exert an epigenetic effect and is a potential bioactive compound for CRC treatment.

Curcumin suppresses cell proliferation and triggers apoptosis in vemurafenib‐resistant melanoma cells by downregulating the EGFR signaling pathway

“…All samples were analyzed using NucleoView NC-3000 software (version 1.4), as previously described. 27,29,30 2.10 | Caspase-9 and caspase-3 activities assay A375.S2/VR cells (1 Â 10 6 cells/well) into 6-well plates were treated with or without 12.5 μM curcumin for 6, 12, and 24 h at 37 C. The cells were harvested and re-suspended with diluted the FLICA reagent and Hoechst 33342, according to the manufacturer's protocol (FAM-FLICA Caspases-3/7 Assay Kit and FAM-FLICA Caspase-9 Assay Kit) for 1 h at 37 C. After washing, cells were resuspended in 100 μl apoptosis wash buffer supplemented with PI. The 30 μl cell suspension was immediately loaded into a 2-chamber slide (NC-Slide A2) for analysis on the NucleoCounter NC-3000 Advanced Image Cytometer using the built-in caspase assay program, as previously described.…”

Back to the Roots—An Overview of the Chemical Composition and Bioactivity of Selected Root-Essential Oils

“…It is stated that the putative chemopreventive mechanism is mainly due to controlling the regulation of estrogen synthesis, the expression of estrogen receptors and cell proliferative markers [ 77 ]. Only recently, a novel mechanism was found by which AITC triggered mitochondrial-dependent apoptosis via G2/M phase arrest and ROS-based ER stress in colorectal adenocarcinoma cells (HT-29 cells) [ 78 ]. Furthermore, results indicated AITC as a potential component worth further investigation in human cisplatin-resistant oral cancer.…”

Differentiated Effects of Allyl Isothiocyanate in Diabetic Rats: From Toxic to Beneficial Action

“…Results of other studies indicate that AITC may induce endoplasmic reticulum stress and mitochondrial damage, which is associated with depletion of intracellular energy. Moreover, AITC may markedly increase formation of reactive oxygen species (ROS) [38]. Glucose-induced insulin secretion is strongly dependent on mitochondrial metabolism and ATP formation [39].…”