Yuan Lingqin, Sheng Xiugui, Clark Leslie H, Zhang Lu, Guo Hui, Jones Hannah M, Willson Adam K, Gehrig Paola A, Zhou Chunxiao, Bae-Jump Victoria L
Department of Gynecologic Oncology, Shandong Cancer Hospital Affiliated to Shandong University, Shandong Academy of Medical SciencesJinan, Shandong, China; Division of Gynecological Oncology, University of North CarolinaChapel Hill, NC, USA.
Department of Gynecologic Oncology, Shandong Cancer Hospital Affiliated to Shandong University, Shandong Academy of Medical Sciences Jinan, Shandong, China.
Am J Transl Res. 2016 Oct 15;8(10):4265-4277. eCollection 2016.
Our overall goal was to investigate the anti-tumor activity of the glutaminase 1 (GLS1) Inhibitor compound 968 in ovarian cancer cells. The human ovarian cancer cell lines, HEY, SKOV3 and IGROV-1 were used. Cell proliferation was assessed by MTT assay after treatment with compound 968. Cell cycle progression and Annexin V expression were evaluated using Cellometer. Western blotting was performed to determine changes in GLS1, cellular stress and cell cycle checkpoints. Reactive oxygen species (ROS) and glutamate dehydrogenase (GDH) activity were assessed by ELISA assay. Compound 968 significantly inhibited cell proliferation and the expression of GLS1 in a dose-dependent manner in all three ovarian cancer cell lines. Compound 968 induced G1 phase cell cycle arrest and apoptosis. Treatment with compound 968 increased ROS levels and induced the protein expression of calnexin, binding immunoglobulin protein (BiP) and protein kinase RNA-like endoplasmic reticulum kinase (PERK). Deprivation of glutamine increased the sensitivity of cells to paclitaxel, and compound 968 sensitized cells to the anti-proliferative effects of paclitaxel. Compound 968 inhibited cell growth in ovarian cancer cells through induction of G1 phase cell cycle arrest, apoptosis and cellular stress, suggesting that targeting GLS1 provide a novel therapeutic strategy for ovarian cancer.
我们的总体目标是研究谷氨酰胺酶1（GLS1）抑制剂化合物968在卵巢癌细胞中的抗肿瘤活性。使用了人卵巢癌细胞系HEY、SKOV3和IGROV-1。用化合物968处理后，通过MTT法评估细胞增殖。使用细胞计数仪评估细胞周期进程和膜联蛋白V表达。进行蛋白质免疫印迹以确定GLS1、细胞应激和细胞周期检查点的变化。通过酶联免疫吸附测定法评估活性氧（ROS）和谷氨酸脱氢酶（GDH）活性。化合物968在所有三种卵巢癌细胞系中均以剂量依赖性方式显著抑制细胞增殖和GLS1的表达。化合物968诱导G1期细胞周期停滞和凋亡。用化合物968处理可提高ROS水平，并诱导钙连接蛋白、结合免疫球蛋白蛋白（BiP）和蛋白激酶RNA样内质网激酶（PERK）的蛋白表达。谷氨酰胺缺乏增加了细胞对紫杉醇的敏感性，并且化合物968使细胞对紫杉醇的抗增殖作用敏感。化合物968通过诱导G1期细胞周期停滞、凋亡和细胞应激来抑制卵巢癌细胞的生长，这表明靶向GLS1为卵巢癌提供了一种新的治疗策略。
