Rong Wang1, Xin Xiao2, Peng-Yuan Wang2, Lin Wang2, Qiunong Guan3, Caigan Du4, Xiao-Juan Wang5. 1. Department of Pharmacy, School of Stomatology, The Fourth Military Medical University, Xi'an, Shaanxi 710032, China; Department of Pharmacology, Xi'an Jiaotong University College of Medicine, Xi'an, Shaanxi 710061, China. 2. Department of Pharmacy, School of Stomatology, The Fourth Military Medical University, Xi'an, Shaanxi 710032, China. 3. Department of Urologic Sciences, University of British Columbia, Vancouver, BC V6H 3Z6, Canada. 4. Department of Urologic Sciences, University of British Columbia, Vancouver, BC V6H 3Z6, Canada. Electronic address: caigan@mail.ubc.ca. 5. Department of Pharmacy, School of Stomatology, The Fourth Military Medical University, Xi'an, Shaanxi 710032, China. Electronic address: wxjyh231@fmmu.edu.cn.
Abstract
AIMS: Ardipusilloside I (ADS-I), a triterpenoid saponin isolated from Ardisia pusilla A.DC (Myrsinaceae), has been recently tested for cancer treatment including brain cancer. However, the mechanism of its action remains elusive. The present study was to investigate the role of autophagy activation in the anti-tumor activities of ADS-I in human glioma cells. MAIN METHODS: The tetrazolium dye (MTT) colorimetric assay was used for the measurement of cell proliferation in cultured glioma cells, transmission electron microscopy (TEM) for the examination of autophagic activity, flow cytometric analysis for the determination of cell cycle and apoptotic cells, and immunocytochemistry and Western blot for protein expression of microtubule-associated protein light-chain 3 (LC3) and Beclin 1. KEY FINDINGS: ADS-I significantly inhibited the proliferation of both U373 and T98G glioma cells in cultures in a dose-dependent manner. The cytotoxic activity of ADS-I against glioma cell growth was associated not only with the induction of cell cycle arrest at G2/M phase and cell apoptosis in flow cytometric analysis, but also with the activation of autophagy, indicated by the formation of autophagosomes and up-regulated expression of both autophagic protein Beclin 1 and LC3 in glioma cells. Additionally, the treatment with chloroquine, an autophagy inhibitor, reduced ADS-1-mediated cell death. SIGNIFICANCE: These data suggest that the anti-proliferative activity of ADS-I in human glioma cells is associated with the activation of autophagy in addition to cell cycle arrest and apoptosis, and the antagonistic effect of chloroquine suggests an important role of autophagy in ADS-I-mediated cell death against tumor growth.
AIMS: Ardipusilloside I (ADS-I), a triterpenoidsaponin isolated from Ardisia pusilla A.DC (Myrsinaceae), has been recently tested for cancer treatment including brain cancer. However, the mechanism of its action remains elusive. The present study was to investigate the role of autophagy activation in the anti-tumor activities of ADS-I in humanglioma cells. MAIN METHODS: The tetrazolium dye (MTT) colorimetric assay was used for the measurement of cell proliferation in cultured glioma cells, transmission electron microscopy (TEM) for the examination of autophagic activity, flow cytometric analysis for the determination of cell cycle and apoptotic cells, and immunocytochemistry and Western blot for protein expression of microtubule-associated protein light-chain 3 (LC3) and Beclin 1. KEY FINDINGS: ADS-I significantly inhibited the proliferation of both U373 and T98G glioma cells in cultures in a dose-dependent manner. The cytotoxic activity of ADS-I against glioma cell growth was associated not only with the induction of cell cycle arrest at G2/M phase and cell apoptosis in flow cytometric analysis, but also with the activation of autophagy, indicated by the formation of autophagosomes and up-regulated expression of both autophagic protein Beclin 1 and LC3 in glioma cells. Additionally, the treatment with chloroquine, an autophagy inhibitor, reduced ADS-1-mediated cell death. SIGNIFICANCE: These data suggest that the anti-proliferative activity of ADS-I in humanglioma cells is associated with the activation of autophagy in addition to cell cycle arrest and apoptosis, and the antagonistic effect of chloroquine suggests an important role of autophagy in ADS-I-mediated cell death against tumor growth.
Authors: Huan Dang; Ji Wang; Jiang-Xue Cheng; Peng-Yuan Wang; Ying Wang; Li-Fei Cheng; Caigan Du; Xiao-Juan Wang Journal: Am J Cancer Res Date: 2014-12-15 Impact factor: 6.166
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