Chuan-bin Yang1, Wei-jing Pei2, Jia Zhao1, Yuan-yuan Cheng1, Xiao-hui Zheng3, Jian-hui Rong1. 1. School of Chinese Medicine, Li Ka Shing Faculty of Medicine, The University of Hong Kong, Pokfulam, Hong Kong, China. 2. RITS and NWU Joint Laboratory for New Drugs Research of TCM, Research Institute of Tsinghua University in Shenzhen, Shenzhen 518000, China. 3. 1] RITS and NWU Joint Laboratory for New Drugs Research of TCM, Research Institute of Tsinghua University in Shenzhen, Shenzhen 518000, China [2] Key Laboratory of Resource Biology and Biotechnology in Western China, School of Life Sciences, Northwest University, Xi'an 710069, China.
Abstract
AIM: The purpose of the present study was to investigate the anticancer activity of bornyl caffeate in the human breast cancer cell line MCF-7. METHODS: The cell viability was determined using the MTT assay, and apoptosis was initially defined by monitoring the morphology of the cell nuclei and staining an early apoptotic biomarker with Annexin V-FITC. The mitochondrial membrane potential was visualized by JC-1 under fluorescence microscopy, whereas intracellular reactive oxygen species (ROS) were assessed by flow cytometry. The expression of apoptosis-associated proteins was determined by Western blotting analysis. RESULTS: Bornyl caffeate induced apoptosis in MCF-7 cells in a dose- and time-dependent manner. Consistently, bornyl caffeate increased Bax and decreased Bcl-xl, resulting in the disruption of MMP and subsequent activation of caspase-3. Moreover, bornyl caffeate triggered the formation of ROS and the activation of the mitogen-activated protein (MAP) kinases p38 and c-Jun N-terminal kinase (JNK). Antioxidants attenuated the activation of MAP kinase p38 but barely affected the activation of JNK. Importantly, the cytotoxicity of bornyl caffeate was partially attenuated by scavenging ROS and inhibited by MAP kinases and caspases. CONCLUSION: The present study demonstrated that bornyl caffeate induced apoptosis in the cancer cell line MCF-7 via activating the ROS- and JNK-mediated pathways. Thus, bornyl caffeate may be a potential anticancer lead compound.
AIM: The purpose of the present study was to investigate the anticancer activity of bornyl caffeate in the humanbreast cancer cell line MCF-7. METHODS: The cell viability was determined using the MTT assay, and apoptosis was initially defined by monitoring the morphology of the cell nuclei and staining an early apoptotic biomarker with Annexin V-FITC. The mitochondrial membrane potential was visualized by JC-1 under fluorescence microscopy, whereas intracellular reactive oxygen species (ROS) were assessed by flow cytometry. The expression of apoptosis-associated proteins was determined by Western blotting analysis. RESULTS:Bornyl caffeate induced apoptosis in MCF-7 cells in a dose- and time-dependent manner. Consistently, bornyl caffeate increased Bax and decreased Bcl-xl, resulting in the disruption of MMP and subsequent activation of caspase-3. Moreover, bornyl caffeate triggered the formation of ROS and the activation of the mitogen-activated protein (MAP) kinases p38 and c-Jun N-terminal kinase (JNK). Antioxidants attenuated the activation of MAP kinase p38 but barely affected the activation of JNK. Importantly, the cytotoxicity of bornyl caffeate was partially attenuated by scavenging ROS and inhibited by MAP kinases and caspases. CONCLUSION: The present study demonstrated that bornyl caffeate induced apoptosis in the cancer cell line MCF-7 via activating the ROS- and JNK-mediated pathways. Thus, bornyl caffeate may be a potential anticancer lead compound.
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