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Autophagy Suppression Accelerates Apoptosis Induced by Norcantharidin in Cholangiocarcinoma

  • Original Article
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Pathology & Oncology Research

Abstract

Norcantharidin is a cantharidin demethylated analog with antitumor effects in many tumors, including cholangiocarcinoma. Autophagy suppression is known to increase chemosensitivity in cholangiocarcinoma. This study aimed to determine whether autophagy suppression accelerates apoptosis induced by norcantharidin in human cholangiocarcinoma cells. The human cholangiocarcinoma cell line QBC939 was incubated in RPMI 1640 medium with or without norcantharidin. Autophagy was induced using HBSS media with Ca2+ and Mg2+ supported by 10 mM HEPES or suppressed by treatment with 3-MA or transfection with siRNA against Atg5. The comparison was drawn between these conditions in mitochondrial membrane potential disturbance, the levels of reactive oxygen species (ROS), apoptotic proteins, and apoptosis. Cholangiocarcinoma cell apoptosis was accelerated by norcantharidin. Autophagy suppression up-regulated norcantharidin’s pro-apoptotic effect, but autophagy induction weakened it. As apoptosis was accelerated, ROS production was up-regulated. Bax protein expression, cytochrome c levels and localization, mitochondrial membrane disturbance, and the levels of caspase-9, caspase-3, and cleaved PARP were higher when autophagy was suppressed, and all of those were down-regulated when autophagy was induced. To sum up, it was found that norcantharidin induced cholangiocarcinoma cell death, and autophagy suppression enhanced the pro-apoptotic action of norcantharidin, which appears to involve the mitochondrial apoptosis pathway activation and ROS generation.

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Abbreviations

LC3-II:

light chain 3-II

NCTD:

norcantharidin

3-MA:

3-methyladenine

HBSS:

Hank’s balanced salt solution

ROS:

reactive oxygen species

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Acknowledgments

The study was financed by grants from the National Science Foundation of China (81670570), Key research and development program of Jiangsu Province (BE2016789).

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Author notes

  1. Yun Wang, Wangjie Jiang, Cunjiang Li and Xuanxuan Xiong contributed equally to this work.

Authors and Affiliations

  1. Key Laboratory of Living Donor Liver Transplantation, Ministry of Public Health, Department of Liver Transplantation Center, The First Affiliated Hospital of Nanjing Medical University, 300 GuangZhou Road, Nanjing, 210029, Jiangsu, China

    Yun Wang, Wangjie Jiang & Xiangcheng Li

  2. Department of Oncological Surgery, Xuzhou City Central Hospital, The Affiliated Hospital of the Southeast University Medical School (Xu zhou), The Tumor Research Institute of the Southeast University (Xu zhou), 199 Jiefang South Road, Xuzhou, 221009, Jiangsu, China

    Yun Wang, Hao Guo & Qingzhong Tian

  3. Department of Thoracic Surgery, Xuzhou City Central Hospital, The Affiliated Hospital of the Southeast University Medical School (Xu zhou), 199 Jiefang South Road, Xuzhou, 221009, Jiangsu, China

    Cunjiang Li

  4. Department of Gastroenterology 2, Xuzhou City Central Hospital, The Affiliated Hospital of the Southeast University Medical School (Xu zhou), 199 Jiefang South Road, Xuzhou, 221009, Jiangsu, China

    Xuanxuan Xiong

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  1. Yun Wang
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Contributions

YW, XL, WJ, CJ and XX conception and design, or analysis and interpretation of data; YW, WJ, CJ, XX, HG and QT drafting the article; XL is the guarantor for the article.

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Correspondence to Xiangcheng Li.

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Wang, Y., Jiang, W., Li, C. et al. Autophagy Suppression Accelerates Apoptosis Induced by Norcantharidin in Cholangiocarcinoma. Pathol. Oncol. Res. 26, 1697–1707 (2020). https://doi.org/10.1007/s12253-019-00719-9

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  • DOI: https://doi.org/10.1007/s12253-019-00719-9

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