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Calpain, Atg5 and Bak play important roles in the crosstalk between apoptosis and autophagy induced by influx of extracellular calcium

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Abstract

Calcium (Ca2+) signals are involved in important checkpoints in cell death pathways and promote both apoptosis and autophagy. However, the relationship between autophagy and apoptosis in response to Ca2+ level elevation is poorly understood. Here, we provided evidence that the influx of extracellular Ca2+ triggered by Trichokonin VI (TK VI), an antimicrobial peptide, induced calpain-dependent apoptosis and autophagy in hepatocellular carcinoma (HCC) cells. Remarkably, TK VI preferentially induced apoptosis that was associated with calpain-mediated Bax and Atg5 cleavage, which resulted in the collapse of the mitochondrial membrane potential and cytochrome c release. Interestingly, truncated, but not full-length Atg5, associated with Bcl-xL and promoted the intrinsic pathway. Moreover, TK VI treatment induced reactive oxygen species (ROS) accumulation, an effect in which Bak might play a major role. This accumulation of ROS resulted in the subsequent disposal of damaged mitochondria within autophagosomes via Atg5-mediated and mitochondria-selective autophagy. Both the inhibition of calpain activity and Bax deficiency activated a switch that promoted an enhancement of autophagy. The inhibition of both apoptosis and autophagy significantly attenuated the TK VI cytotoxicity, indicating that the two processes had stimulatory effects during TK VI-meditated cell death. These results suggested that calpain, Bak and Atg5 were molecular links between autophagy and apoptosis and revealed novel aspects of the crosstalk between these two processes. The potential of TK VI is proposed as a promising anticancer agent for its well-characterized activity of Ca2+ agonist and as a possible novel therapeutic strategy that acts on cancer cell mitochondria.

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Acknowledgments

We thank Dr. Xiao-Yan Li for providing a vector containing cleaved atg5 gene (Academy of Military Medical Sciences, P. R. China). We are grateful to Dr. Adam Salmon (University of Texas Health Science Center at San Antonio) for his critical revision on this manuscript. The work was supported by Hi-Tech Research and Development program of China [2011AA090704], National Natural Science Foundation of China [30870047, 81071804]; Specialized Research Fund for the Doctoral Program of Higher Education [BA2009YY009]; Program of Shandong for Taishan Scholars, Natural Science Foundation of Shandong Province [JQ200910, 2008BS02019]; and Independent Innovation Foundation of Shandong University [2009TS079, 2011DX002].

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The authors declare that they have no conflict of interest.

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Authors and Affiliations

  1. Marine Biotechnology Research Center, State Key Laboratory of Microbial Technology, Shandong University, Jinan, 250100, People’s Republic of China

    Mei Shi, Tian Zhang, Yan Luo, De-Hua Liu, Shu-Tao Xie, Xiao-Yan Song, Xiu-Lan Chen, Bai-Cheng Zhou & Yu-Zhong Zhang

  2. Biological Imaging Center, Institute of Biophysics, Chinese Academy of Sciences, Beijing, 100101, People’s Republic of China

    Lei Sun

  3. Shandong Cancer Hospital, Jinan, 250117, People’s Republic of China

    Guo-Fan Wang

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  1. Mei Shi
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Shi, M., Zhang, T., Sun, L. et al. Calpain, Atg5 and Bak play important roles in the crosstalk between apoptosis and autophagy induced by influx of extracellular calcium. Apoptosis 18, 435–451 (2013). https://doi.org/10.1007/s10495-012-0786-2

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