PSNA College of Engineering and Technology, Kothhandaraman Nagar, Dindugal624 622, Tamil Nadu, India.
Cell Biol Int. 2011 Jun;35(6):607-15. doi: 10.1042/CBI20100118.
Eugenol, a natural compound available in honey and various plants extracts including cloves and Magnoliae flos, is exploited for various medicinal applications. Since most of the drugs used in the cancer are apoptotic inducers, the apoptotic effect and anticancer mechanism of eugenol were investigated against colon cancer cells. Antiproliferative effect was estimated using MTT [3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide assay]. Earlier events like MMP (mitochondrial membrane potential), thiol depletion and lipid layer break were measured by using flow cytometry. Apoptosis was evaluated using PI (propidium iodide) staining, TUNEL (terminal deoxynucleotidyl transferase-mediated dUTP nick end labelling) assay and DNA fragmentation assay. MTT assay signified the antiproliferative nature of eugenol against the tested colon cancer cells. PI staining indicated increasing accumulation of cells at sub-G1-phase. Eugenol treatment resulted in reduction of intracellular non-protein thiols and increase in the earlier lipid layer break. Further events like dissipation of MMP and generation of ROS (reactive oxygen species) were accompanied in the eugenol-induced apoptosis. Augmented ROS generation resulted in the DNA fragmentation of treated cells as shown by DNA fragmentation and TUNEL assay. Further activation of PARP (polyadenosine diphosphate-ribose polymerase), p53 and caspase-3 were observed in Western blot analyses. Our results demonstrated molecular mechanism of eugenol-induced apoptosis in human colon cancer cells. This research will further enhance eugenol as a potential chemopreventive agent against colon cancer.
丁香酚是一种天然化合物，存在于蜂蜜和各种植物提取物中，包括丁香和 Magnoliae flos，它被用于各种医学应用。由于大多数用于癌症的药物都是凋亡诱导剂，因此研究了丁香酚对结肠癌细胞的凋亡作用和抗癌机制。使用 MTT [3-(4,5-二甲基噻唑-2-基)-2,5-二苯基四唑溴盐测定法] 估计增殖抑制作用。使用流式细胞术测量早期事件，如 MMP（线粒体膜电位）、巯基耗竭和脂质层破裂。使用 PI（碘化丙啶）染色、TUNEL（末端脱氧核苷酸转移酶介导的 dUTP 缺口末端标记）测定和 DNA 片段化测定评估凋亡。MTT 测定表明丁香酚对测试的结肠癌细胞具有增殖抑制作用。PI 染色表明细胞在亚 G1 期的积累增加。丁香酚处理导致细胞内非蛋白巯基减少和早期脂质层破裂增加。在丁香酚诱导的凋亡中还伴随着其他事件，如 MMP 耗散和 ROS（活性氧）的产生。增强的 ROS 生成导致处理细胞的 DNA 片段化，如 DNA 片段化和 TUNEL 测定所示。在 Western blot 分析中观察到 PARP（多聚腺苷二磷酸核糖聚合酶）、p53 和 caspase-3 的进一步激活。我们的结果表明了丁香酚诱导人结肠癌细胞凋亡的分子机制。这项研究将进一步增强丁香酚作为预防结肠癌的潜在化学预防剂的作用。
