7 Methyl indole ethyl isothiocyanate causes ROS mediated apoptosis and cell cycle arrest in endometrial cancer cells
7 Methyl indole ethyl isothiocyanate causes ROS mediated apoptosis and cell cycle arrest in endometrial cancer cells
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DOI：
10.1016/j.ygyno.2012.04.041
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发表时间：
2012-08-01
影响因子：
4.7
通讯作者：
Singh, Rakesh K.
中科院分区：
文献类型：
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作者：
Kristjansdottir, Katrin;Kim, Kyukwang;Singh, Rakesh K.
Objective.晚期子宫内膜癌的化疗选择有限，迫切需要更新的治疗药物。本研究旨在探讨7-甲基吲哚乙基异硫氰酸酯（7 Me-IEITC）对子宫内膜癌细胞系的治疗作用。本实验室合成了7 Me-IEITC。MTS法检测7 Me-IEITC处理后ECC-1和KLE子宫内膜癌细胞的细胞活力。DAPI染色和TUNEL法进一步证实了细胞形态和凋亡。流式细胞仪测定细胞内活性氧（ROS）、线粒体跨膜去极化电位（Δ Psim）和细胞周期时相。蛋白质印迹法分析凋亡、存活和细胞周期进程相关蛋白的表达。7 Me-IEITC以剂量依赖性方式降低ECC-1和KLE癌细胞系的生存力（IC 50类似于2.5-10 μ M）。7 Me-IEITC处理引起子宫内膜癌KLE和ECC-1细胞线粒体跨膜电位降低，ROS产生增加，导致细胞凋亡激活。7 Me-IEITC处理激活Bad，抑制Bcl 2磷酸化，随后PARP-1失活以及caspase 3和7激活。7 Me-IEITC处理KLE细胞后，CDC 25和cyclin-D1表达下调，细胞周期阻滞于S期。抗坏血酸预处理可抑制7 Me-IEITC诱导的ECC-1和KLE细胞凋亡，提示7 Me-IEITC主要通过产生ROS介导细胞毒作用。7 Me-IEITC在子宫内膜癌细胞系模型中显示出有希望的细胞毒作用。(C)2012 Elsevier Inc. All rights reserved.
Objective. Chemotherapy options for advanced endometrial cancer are limited and newer therapeutic agents are urgently needed. This study describes the therapeutic potential of 7 Methyl-indole ethyl isothiocyanate (7Me-IEITC) in endometrial cancer cell lines.Methods. 7Me-IEITC was synthesized in our laboratory. The cell viability of 7Me-IEITC treated ECC-1 and KLE endometrial cancer cell was determined by MTS assay. Morphology and apoptosis were further confirmed by DAPI-staining and TUNEL assay. The measurement of reactive oxygen species (ROS), mitochondrial transmembrane depolarization potential (Delta Psi m) and cell cycle phase was determined by FACS analysis. Expression of proteins involved in apoptosis, survival and cell-cycle progression was analyzed by Western blotting.Results. 7Me-IEITC reduced the viability of the ECC-1 and KLE cancer cell-lines (IC50 similar to 2.5-10 mu M) in a dose dependent fashion. 7Me-IEITC treatment caused mitochondrial transmembrane potential reduction, elevated the production of ROS, leading to activation of apoptosis in endometrial cancer KLE and ECC-1 cells. 7Me-IEITC treatment activated Bad, suppressed Bcl2 phosphorylation followed by PARP-1 deactivation and caspase 3 and 7 activation. 7Me-IEITC treatment arrested the progression of KLE cells in S-phase and caused CDC25 and cyclin-D1 downregulation. Pre-treatment with ascorbic acid abrogated 7Me-IEITC induced apoptosis in ECC-1 and KLE cells, suggesting that 7Me-IEITC mediated cytotoxicity is primarily through ROS production.Conclusion. 7Me-IEITC demonstrated promising cytotoxic effects in endometrial cancer cell line model. (C) 2012 Elsevier Inc. All rights reserved.