Hoechst 33342-induced autophagy protected HeLa cells from caspase-independent cell death with the participation of ROS |
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Authors: | Zheng Fang Yang Wen-Jun Sun Ke-Jing Wan Xiao-Mei Man Na Wen Long-Ping |
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Institution: | Hefei National Laboratory for Physical Sciences at Microscale & School of Life Sciences, University of Science and Technology of China, Anhui, China. |
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Abstract: | Autophagy, an evolutionarily-conserved intracellular organelle and protein degradation process, may exhibit drastically different effects on cell survival depending on the particular environmental and culturing conditions. Hoechst 33342 (HO), a fluorescent dye widely used for staining DNA, has been reported to induce apoptosis in mammalian cells. Here we showed that, in addition to caspase-independent cell death, HO also induced autophagy in HeLa cells, as evidenced by the accumulation of autophagosomes, LC3 form conversion and LC3 puncta formation in a cell line stably expressing GFP-LC3. HO treatment led to generation of reactive oxygen species (ROS), and inhibition of ROS with N-acetyl-l-cysteine (NAC) abrogated both autophagy and caspase-independent cell death. Finally, autophagy played a protective role against caspase-independent cell death, as cell death induced by HO was enhanced under pharmacological and siRNA-mediated genetic inhibition of autophagy. |
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