Journal of Korean Medical Science, 2013, 28(4):527-33.

The Synergistic Apoptotic Interaction of Indole-3-Carbinol and Genistein with TRAIL on Endometrial Cancer Cells

Bidur Parajuli, et al

Department of Obstetrics and Gynecology, Keimyung University, School of Medicine, Daegu, Korea; Institute for Cancer Research, Keimyung University, School of Medicine, Daegu, Korea; Department of Oncology, Lombardi Cancer Center, Georgetown University, Washington DC, USA

Induction of apoptosis in target cells is a key mechanism by which chemotherapy promotes cell killing. The purpose of this study was to determine whether Indole-3-Carbinol (I3C) and Genistein in combination with tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) induce apoptosis in endometrial cancer cell (Ishikawa) and to assess apoptotic mechanism. The MTT assay and flow cytometry were performed to determine cell viability and cell cycle. The induction of apoptosis was measured by caspase-3 activity test, DNA fragmentation assay, annexin V binding assay and western blot analysis. There was no effect in cell growth inhibition and cell cycle progression alone or in two-combination. However, the treatment of I3C and Genistein followed by TRAIL showed significant cell death and marked increase in sub-G1 arrest. Three-combination treatment revealed elevated expression of DR4, DR5 and cleaved forms of caspase-3, caspase-8, PARP. The Flip was found down regulated. Moreover, increase in caspase-3 activity and DNA fragmentation indicated the induction of apoptosis. The results indicate that I3C and Genistein with TRAIL synergistically induced apoptosis via death receptor dependent pathway. Our findings might provide a new insight into the development of novel combination therapies against endometrial cancer.

韩国《韩国医学科学杂志》，2013年4月

吲哚-3-甲醇和金雀异黄素对子宫内膜癌细胞的协同凋亡相互作用

比德 帕拉久利，等

韩国大邱启明大学医学院妇产科和癌症研究所；美国华盛顿乔治敦大学隆巴迪癌症中心肿瘤科

诱导靶向细胞凋亡是化疗促进细胞杀伤力的关键机制。本研究的目的是确认吲哚-3-原醇 (I3C) 和金雀异黄素与肿瘤坏死因子相关凋亡诱导配体 (TRAIL) 联合是否诱导子宫内膜癌细胞 （Ishikawa）凋亡，并评估细胞凋亡机制。用MTT 法和流式细胞术来确定细胞存活率和细胞周期。用半胱氨酸蛋白酶-3 活性试验、 DNA碎片测验、 膜联蛋白 V 绑定分析法和免疫印迹分析测定诱导的细胞凋亡。单独或两者组合疗法对细胞生长抑制和细胞周期进程都没有作用。然而，在TRAIL之后的I3C 和金雀异黄素的治疗表现出明显的细胞死亡和sub-G1阻的标记性提高。三组合疗法表现出DR4、 DR5和解离的半胱氨酸蛋白酶-3，半胱氨酸蛋白酶-8，PARP被提高的表达。发现Filp被下调。此外，增加的半胱氨酸蛋白酶-3 活性和 DNA 碎片表明细胞凋亡的诱导。结果表明 I3C 和金雀异黄素与TRAIL协同通过死亡受体依赖途径诱导细胞凋亡。我们的研究结果可能提供了对子宫内膜癌新型组合疗法的新见解。