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Angelicin induces apoptosis through intrinsic caspase-dependent pathway in human SH-SY5Y neuroblastoma cells

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Abstract

Angelicin is structurally related to psoralens, a well-known chemical class of photosensitizers used for its antiproliferative activity in treatment of different skin diseases. To verify the activity of angelicin, we employed human SH-SY5Y neuroblastoma cells to investigate its cytotoxicity, although its mechanism of action has not yet been fully elucidated. Here, we examined the cellular cytotoxicity of angelicin by cell viability assay, DNA fragmentation by DNA ladder assay, and activation of caspases and Bcl-2 family proteins by western blot analyses. The results of our investigation suggest that angelicin increased cellular cytotoxicity in a dose- and time-dependent manner with IC50 of 49.56 μM at 48 h of incubation. In addition, angelicin dose-dependently downregulated the expression of anti-apoptotic proteins including Bcl-2, Bcl-xL, and Mcl-1 suggesting the involvement of the intrinsic mitochondria-mediated apoptotic pathway which did not participate in Fas/FasL-induced caspase-8-mediated extrinsic, MAP kinases, and PI3K/AKT/GSK-3β pathway. Furthermore, we clarified the dose-dependent upregulation of caspase-9 and caspase-3 which indicated that angelicin-induced apoptosis is mediated primarily through the intrinsic caspase-mediated pathway. In particular, the caspase-3 inhibitor, DEVD-fmk, induced a reduction in angelicin-induced cytotoxicity which confirmed that the intrinsic caspase-dependent pathway during this apoptosis which did not prevent cytotoxicity using MAP kinases and GSK-3 inhibitor. Taken together, our data shows that angelicin is an effective apoptosis-inducing natural compound of human SH-SY5Y neuroblastoma cells which suggests that this compound may have a role in future therapies for human neuroblastoma cancer.

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Acknowledgments

We thank Dr. Jae-Yong Lee for providing us with the human SH-SY5Y neuroblastoma cell line. This research was supported by a grant of the Korea Healthcare Technology R&D Project, Ministry of Health & Welfare (A090792) and Basic Science Research Program through the Natural Research Foundation of Korea (NRF) funded by the Ministry of Education, Science and Technology (2011-0003931, 2011-0005281), the Republic of Korea.

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  1. Department of Pharmacology, Institute of Natural Medicine, College of Medicine, Hallym University, Chuncheon, Gangwon-do, 200-702, South Korea

    Md. Ataur Rahman, Nam-Ho Kim, Haijie Yang & Sung-Oh Huh

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  1. Md. Ataur Rahman
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  3. Haijie Yang
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Correspondence to Sung-Oh Huh.

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Ataur Rahman, M., Kim, NH., Yang, H. et al. Angelicin induces apoptosis through intrinsic caspase-dependent pathway in human SH-SY5Y neuroblastoma cells. Mol Cell Biochem 369, 95–104 (2012). https://doi.org/10.1007/s11010-012-1372-1

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  • DOI: https://doi.org/10.1007/s11010-012-1372-1

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