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A possible cross-talk between autophagy and apoptosis in generating an immune response in melanoma

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Abstract

Melanoma is the most aggressive form of skin cancer, responsible for the majority of skin cancer related deaths. Thus, the search for natural molecules which can effectively destroy tumors while promoting immune activation is essential for designing novel therapies against metastatic melanoma. Here, we report for the first time that a natural triterpenoid, Ganoderic acid DM (GA-DM), induces an orchestrated autophagic and apoptotic cell death, as well as enhanced immunological responses via increased HLA class II presentation in melanoma cells. Annexin V staining and flow cytometry showed that GA-DM treatment induced apoptosis of melanoma cells, which was supported by a detection of increased Bax proteins, co-localization and elevation of Apaf-1 and cytochrome c, and a subsequent cleavage of caspases 9 and 3. Furthermore, GA-DM treatment initiated a possible cross-talk between autophagy and apoptosis as evidenced by increased levels of Beclin-1 and LC3 proteins, and their timely interplay with apoptotic and/or anti-apoptotic molecules in melanoma cells. Despite GA-DM’s moderate cytotoxicity, viable cells expressed high levels of HLA class II proteins with improved antigen presentation and CD4+ T cell recognition. The antitumor efficacy of GA-DM was also investigated in vivo in murine B16 melanoma model, where GA-DM treatment slowed tumor formation with a significant reduction in tumor volume. Taken together, these findings demonstrate the potential of GA-DM as a natural chemo-immunotherapeutic capable of inducing a possible cross-talk between autophagy and apoptosis, as well as improved immune recognition for sustained melanoma tumor clearance.

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Acknowledgments

This work was supported by grants from the National Institutes of Health (R01 CA129560 and R01 CA129560-S1 to A. Haque). The research presented in this article was also supported in part by the Tissue Biorepository and Flow Cytometry Shared Resource as part of the Hollings Cancer Center at the Medical University of South Carolina which is funded by a Cancer Center Support Grant P30 CA138313. We thank Drs. Janice Blum (Indiana University, Indianapolis), Craig Slingluff Jr. (University of Virginia, Charlottesville) for melanoma cell lines; Drs. L. Xiang, Christina Johnson, and Carola Neumann (MUSC) for antibodies; Dr. Sakamuri Reddy (Dept. of Pediatrics, MUSC) for his critical reading of the manuscript and Dr. M. Rubinstein (Dept. of Surgery) for technical assistance. We also gratefully acknowledge Dr. Chenthamarakshan Vasu (MUSC) for his Fow Cytometry Facility.

Conflicts of Interest

No conflicts of interest were disclosed by the authors.

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Authors and Affiliations

  1. Department of Microbiology and Immunology, Medical University of South Carolina, 173 Ashley Avenue, BSB-201, Charleston, SC, 29425, USA

    Azim Hossain, Faisal F. Y. Radwan, Bently P. Doonan, Jason M. God, Lixia Zhang & Azizul Haque

  2. Hollings Cancer Center, Medical University of South Carolina, 173 Ashley Avenue, Charleston, SC, 29425, USA

    Azim Hossain, Faisal F. Y. Radwan, Bently P. Doonan, Jason M. God, Lixia Zhang & Azizul Haque

  3. Children’s Research Institute, Medical University of South Carolina, 173 Ashley Avenue, Charleston, SC, 29425, USA

    Azim Hossain, Faisal F. Y. Radwan, Bently P. Doonan, Jason M. God, Lixia Zhang, P. Darwin Bell & Azizul Haque

  4. Division of Nephrology, Medical University of South Carolina, 173 Ashley Avenue, Charleston, SC, 29425, USA

    P. Darwin Bell

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  1. Azim Hossain
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  2. Faisal F. Y. Radwan
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Correspondence to Azizul Haque.

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Azim Hossain and Faisal Radwan contributed equally to this study.

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Hossain, A., Radwan, F.F.Y., Doonan, B.P. et al. A possible cross-talk between autophagy and apoptosis in generating an immune response in melanoma. Apoptosis 17, 1066–1078 (2012). https://doi.org/10.1007/s10495-012-0745-y

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