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AMID Mediates Adenosine-Induced Caspase-Independent HuH-7 Cell ApoptosisYang D.1,2 · Yaguchi T.2 · Nagata T.2 · Gotoh A.3 · Dovat S.4 · Song C.4 · Nishizaki T.2
1Laboratory of Molecular Cell Biology, Institute of Biochemistry and Cell Biology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, Shanghai,2Division of Bioinformation, Department of Physiology, Hyogo College of Medicine, Mukogawa-cho,3Laboratory of Cell and Gene Therapy, Institute for Advanced Medical Sciences, Hyogo College of Medicine, Mukogawa-cho,4Departments of Pediatrics, University of Wisconsin-Madison Medical School, Madison Corresponding Author
Prof. Tomoyuki Nishizaki
Division of Bioinformation, Dept of Physiology, Hyogo College of Medicine,
1-1 Mukogawa-cho, Nishinomiya 663-8501 (Japan)
Tel. +81-798-45-6397, Fax +81-798-45-6649
Background/Aims: The mechanism underlying extracellular adenosine-induced caspase-independent apoptosis in HuH-7 human hepatoma cells is not fully understood. The present study investigated the role for apoptosis-inducing factor (AIF)-homologous mitochondrion-associated inducer of death (AMID) in the pathway. Methods: To see the implication of AMID in adenosine-induced HuH-7 cell apoptosis, real-time reverse transcription-polymerase chain reaction (RT-PCR), immunofluorescent cytochemistry, time-laps GFP monitoring, cell cycle analysis, flow cytometry, Western blotting, cell viability assay, and TUNEL staining were carried out. Results: Adenosine upregulated AMID expression in HuH-7 cells, and translocated AMID from the cytosol into the nucleus. Adenosine induced HuH-7 cell apoptosis, and the effect was further enhanced by overexpressing AMID. Adenosine-induced HuH-7 cell apoptosis, alternatively, was inhibited by knocking-down AMID. Conclusion: The results of the present study provide evidence for AMID as a critical factor for adenosine-induced caspase-independent HuH-7 cell apoptosis.
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