School of Pharmacy, Shihezi University, 832002, Shihezi, P.R. China.
Recent Pat Anticancer Drug Discov. 2013 May;8(2):191-9.
Isoliquiritigenin (ISL), a licorice chalconoid, is a bioactive agent with chemopreventive potential that has been patented for tumor treatment in China. This study investigated the mechanisms of ISL-induced apoptosis in ovarian carcinoma SKOV-3 cells. Cell viability was evaluated using a 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-tetrazolium bromide assay. The apoptotic rate was determined via flow cytometry using an annexin V-FITC apoptosis detection kit. The intracellular reactive oxygen species (ROS) levels were assessed using a 2,7-dichlorofluorescein probe assay. Malondialdehyde (MDA) formation was determined via thiobarbituric acid reactive substance test. The expressions of growth arrest and DNA damage-inducible gene (GADD153/CHOP), 78 kDa glucose-regulated protein (GRP 78), α-subunit of eukaryotic initiation factor 2 (eIF2α) phosphorylation, activating transcription factor 6α (ATF6α), and unspliced form of X-box binding protein1 (XBP1U) were analyzed via Western blot. Caspase-3 and caspase-12 activities were assessed using a fluorometric kit. Findings indicate that ISL significantly inhibits SKOV-3 cell proliferation, increases intracellular ROS levels, and causes SKOV-3 cell apoptosis. Moreover, ISL-exposed SKOV-3 cells trigger endoplasmic reticulum (ER) stress, as indicated by the enhancement of ER stress-related molecules p-eIF2α, GADD153/CHOP, GRP78, XBP1 expression, and cleavage of ATF6α. However, caspase-12 inhibitor (Z-ATAD) effectively and partially prevents ROS and MDA formation and inhibits ISL-induced SKOV-3 cell apoptosis. ISL induces apoptosis via ER stress-triggered signaling pathways in SKOV-3 cells. ER stress-induced cancer cell apoptosis has been discussed in some patents.
异甘草素（ISL）是一种甘草查尔酮，具有化学预防潜力，已在中国获得专利用于肿瘤治疗。本研究探讨了 ISL 诱导卵巢癌 SKOV-3 细胞凋亡的机制。通过 3-(4,5-二甲基噻唑-2-基)-2,5-二苯基四氮唑溴化物测定法评估细胞活力。通过 Annexin V-FITC 凋亡检测试剂盒通过流式细胞术测定凋亡率。通过 2,7-二氯荧光素探针测定法评估细胞内活性氧（ROS）水平。通过硫代巴比妥酸反应物质试验测定丙二醛（MDA）形成。通过 Western blot 分析生长停滞和 DNA 损伤诱导基因（GADD153/CHOP）、78 kDa 葡萄糖调节蛋白（GRP 78）、真核起始因子 2（eIF2α）磷酸化的 α 亚基、激活转录因子 6α（ATF6α）和未剪接的 X 盒结合蛋白 1（XBP1U）的表达。使用荧光法试剂盒评估 caspase-3 和 caspase-12 的活性。结果表明，ISL 显著抑制 SKOV-3 细胞增殖，增加细胞内 ROS 水平，并导致 SKOV-3 细胞凋亡。此外，ISL 暴露的 SKOV-3 细胞引发内质网（ER）应激，如 ER 应激相关分子 p-eIF2α、GADD153/CHOP、GRP78、XBP1 表达和 ATF6α 的切割增强所示。然而，caspase-12 抑制剂（Z-ATAD）有效且部分地阻止 ROS 和 MDA 的形成并抑制 ISL 诱导的 SKOV-3 细胞凋亡。ISL 通过 ER 应激触发的信号通路在 SKOV-3 细胞中诱导细胞凋亡。一些专利讨论了 ER 应激诱导的癌细胞凋亡。
