Sensitivity of allyl isothiocyanate to induce apoptosis via ER stress and the mitochondrial pathway upon ROS production in colorectal adenocarcinoma cells
Sensitivity of allyl isothiocyanate to induce apoptosis via ER stress and the mitochondrial pathway upon ROS production in colorectal adenocarcinoma cells
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DOI：
10.3892/or.2020.7700
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发表时间：
2020-10-01
期刊：
影响因子：
4.2
通讯作者：
Yang, Jai-Sing
中科院分区：
文献类型：
--
作者：
Chiang, Jo-Hua;Tsai, Fuu-Jen;Yang, Jai-Sing
异硫氰酸烯丙酯（AITC）是一种具有生物活性的植物化学物质，是十字花科蔬菜的主要成分，具有良好的化学预防和抗癌作用。然而，关于AITC在体外诱导结直肠癌（CRC）细胞凋亡方面发挥抗肿瘤作用的报道并不清楚。本研究旨在探讨AITC诱导人大肠癌HT-29细胞凋亡的内质网应激机制。我们的研究结果表明，AITC减少细胞生长和数量，降低活力，并促进凋亡细胞死亡的形态学变化。流式细胞术DNA分析显示G2/M期阻滞，通过蛋白质印迹分析检测AITC引起的调节蛋白水平的改变。AITC还触发了重要的内在凋亡因子（半胱天冬酶-9/半胱天冬酶-3活性），破坏了线粒体膜电位，并刺激了与细胞相关的凋亡分子（例如，细胞色素c、凋亡蛋白酶激活因子1、凋亡诱导因子和内切核酸酶G）。此外，AITC促进诱导的胞质Ca 2+释放和Ca 2+依赖性ER应激相关信号，如钙蛋白酶1、激活转录因子6 α、葡萄糖调节蛋白78和94、生长停滞和DNA损伤诱导蛋白153（GADD 153）和半胱天冬酶-4。活性氧（ROS）的产生水平被发现诱导ER应激的标志GADD 153，促凋亡标志物caspase-3，和钙蛋白酶活性AITC治疗后。我们的研究结果首次表明，AITC诱导HT-29细胞的G2/M期阻滞和凋亡死亡通过ROS为基础的ER应激和内源性途径（细胞依赖性）。总体而言，AITC可能发挥表观遗传效应，是治疗CRC的潜在生物活性化合物。
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 Ca2+ release and Ca2+-dependent ER stress-related signals, such as calpain 1, activating transcription factor 6 alpha, 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.