Man Wu1, Yuanzhi Lao1, Naihan Xu2, Xiaoyu Wang1, Hongsheng Tan1, Wenwei Fu1, Zhixiu Lin3, Hongxi Xu4. 1. School of Pharmacy, Shanghai University of Traditional Chinese Medicine, Shanghai, 201203, P.R. China; Engineering Research Center of Shanghai Colleges for TCM New Drug Discovery, Shanghai, 201203, P.R. China. 2. Key Lab in Healthy Science and Technology, Division of Life Science, Graduate School at Shenzhen, Tsinghua University, Shenzhen, 518055, P.R. China. 3. Faculty of Science, School of Chinese Medicine, The Chinese University of Hong Kong, Shatin, N.T., Hong Kong SAR, China. 4. School of Pharmacy, Shanghai University of Traditional Chinese Medicine, Shanghai, 201203, P.R. China; Engineering Research Center of Shanghai Colleges for TCM New Drug Discovery, Shanghai, 201203, P.R. China. Electronic address: xuhongxi88@gmail.com.
Abstract
BACKGROUND: Medicinal plants have long been an excellent source of pharmaceutical agents. Autophagy, a catabolic degradation process through lysosomes, plays an important role in tumorigenesis and cancer therapy. PURPOSE: Through a screen designed to identify autophagic regulators from a library of natural compounds, we found that Guttiferone K (GUTK) can activate autophagy in several cancer cell lines. The objective of this study is to investigate the mechanism by which GUTK sensitizes cancer cells to cell death in nutrient starvation condition. METHODS: Cell death analysis was performed by propidium iodide staining with flow cytometry or Annexin V-FITC/PI staining assay. DCFH-DA staining was used for intracellular ROS measurement. Protein levels were analyzed by western blot analysis. Cell viability was measured by MTT assay. RESULTS: Exposure to GUTK was observed to markedly induce GFP-LC3 puncta formation and activate the accumulation of LC3-II and the degradation of p62 in HeLa cells, suggesting that GUTK is an autophagy inducer. Importantly, hydroxychloroquine, an autophagy inhibitor, was found to significantly prevent GUTK-induced cell death in nutrient starvation conditions, suggesting that the cell death observed is largely dependent on autophagy. We further provide evidence that GUTK inhibits Akt phosphorylation, thereby inhibiting the mTOR pathway in cancer cells during nutrient starvation. In addition, GUTK causes the accumulation of reactive oxygen species (ROS) and the phosphorylation of JNK in EBSS, which may mediate both autophagy and apoptosis. CONCLUSION: These data indicate that GUTK sensitizes cancer cells to nutrient stress-induced cell death though Akt/mTOR dependent autophagy pathway.
BACKGROUND: Medicinal plants have long been an excellent source of pharmaceutical agents. Autophagy, a catabolic degradation process through lysosomes, plays an important role in tumorigenesis and cancer therapy. PURPOSE: Through a screen designed to identify autophagic regulators from a library of natural compounds, we found that Guttiferone K (GUTK) can activate autophagy in several cancer cell lines. The objective of this study is to investigate the mechanism by which GUTK sensitizes cancer cells to cell death in nutrient starvation condition. METHODS: Cell death analysis was performed by propidium iodide staining with flow cytometry or Annexin V-FITC/PI staining assay. DCFH-DA staining was used for intracellular ROS measurement. Protein levels were analyzed by western blot analysis. Cell viability was measured by MTT assay. RESULTS: Exposure to GUTK was observed to markedly induce GFP-LC3 puncta formation and activate the accumulation of LC3-II and the degradation of p62 in HeLa cells, suggesting that GUTK is an autophagy inducer. Importantly, hydroxychloroquine, an autophagy inhibitor, was found to significantly prevent GUTK-induced cell death in nutrient starvation conditions, suggesting that the cell death observed is largely dependent on autophagy. We further provide evidence that GUTK inhibits Akt phosphorylation, thereby inhibiting the mTOR pathway in cancer cells during nutrient starvation. In addition, GUTK causes the accumulation of reactive oxygen species (ROS) and the phosphorylation of JNK in EBSS, which may mediate both autophagy and apoptosis. CONCLUSION: These data indicate that GUTK sensitizes cancer cells to nutrient stress-induced cell death though Akt/mTOR dependent autophagy pathway.
Authors: Monika Bhardwaj; Souren Paul; Rekha Jakhar; Imran Khan; Ji In Kang; Ho Min Kim; Jong Won Yun; Seon-Jin Lee; Hee Jun Cho; Hee Gu Lee; Sun Chul Kang Journal: Oncotarget Date: 2017-08-10
Authors: Betty Y K Law; Simon W F Mok; Juan Chen; Francesco Michelangeli; Zhi-Hong Jiang; Yu Han; Yuan Q Qu; Alena C L Qiu; Su-Wei Xu; Wei-Wei Xue; Xiao-Jun Yao; Jia Y Gao; Masood-Ul-Hassan Javed; Paolo Coghi; Liang Liu; Vincent K W Wong Journal: Front Pharmacol Date: 2017-06-16 Impact factor: 5.810