期刊
FREE RADICAL RESEARCH
卷 46, 期 6, 页码 740-749出版社
INFORMA HEALTHCARE
DOI: 10.3109/10715762.2012.670701
关键词
Hoechst 33342; autophagy; caspase-independent cell death; ROS
资金
- Chinese Ministry of Sciences [2007CB935800, 2010CB912804]
- Natural Science Foundation of China [30721002, 31071211, 30830036, 31170966, 31101020]
- Chinese Academy of Sciences [KSCX2-YW-R-139]
- Fundamental Research Funds for the Central Universities [WK2070000008]
- Scientific and Technological Major Special Project [2009ZX09103-650]
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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