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Icaritin activates JNK-dependent mPTP necrosis pathway in colorectal cancer cells

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Tumor Biology

Abstract

The colorectal cancer (CRC) is one leading contributor of cancer-related mortality worldwide. The search for effective anti-CRC agents is valuable. In the current study, we showed that icaritin (ICT), an active natural ingredient from the Chinese plant Epimedium, potently inhibited proliferation and survival of established (HT-29, HCT-116, DLD-1, and SW-620) and primary (patient-derived) CRC cells. Significantly, ICT mainly induced necrosis, but not apoptosis, in CRC cells. The necrosis inhibitor necrostatin-1 attenuated ICT-mediated cytotoxicity in CRC cells. We showed that ICT treatment in CRC cells induced mitochondrial permeability transition pore (mPTP) opening, which was evidenced by mitochondrial membrane potential (MMP) decrease and mitochondrial adenine nucleotide translocator-1 (ANT-1)-cyclophilin-D (CyPD) association. On the other hand, mPTP blockers, including sanglifehrin A, cyclosporin A, and bongkrekic acid, as well as siRNA-mediated knockdown of mPTP component (CyPD or ANT-1), significantly alleviated ICT-mediated cytotoxicity against CRC cells. We suggested that Jun-N-terminal kinase (JNK) activation by ICT mediated mPTP opening and subsequent CRC cell necrosis. JNK pharmacological inhibition, dominant negative mutation, or shRNA downregulation suppressed ICT-induced MMP reduction and subsequent HT-29 cell necrosis. In vivo, oral gavage of ICT dramatically inhibited HT-29 xenograft growth in nude mice. The in vivo activity by ICT was largely attenuated by co-administration with the mPTP blocker CsA. Collectively, our results showed that ICT exerts potent inhibitory effect against CRC cells in vitro and in vivo. JNK-dependent mPTP necrosis pathway could be key mechanism responsible for ICT’s actions.

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Acknowledgments

This work was supported by the National Natural Science Foundation.

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Authors and Affiliations

  1. Department of Interventional Radiology, WuJiang Hospital Affiliated to Nantong University, No. 169 Park Road, Songling Town, WuJiang, Suzhou, 215200, China

    Chunxian Zhou, Hao Wu, Qunying Yang & Dongfeng Xu

  2. Department of General Surgery, The Second Affiliated Hospital of Soochow University, Suzhou, 215000, China

    Zhengrong Chen

  3. Department of Hepatobiliary Surgery of Suzhou Municipal Hospital, Suzhou, 215000, China

    Xingsheng Lu

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  1. Chunxian Zhou
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Correspondence to Dongfeng Xu.

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Chunxian Zhou, Zhengrong Chen, and Xingsheng Lu are co-first authors.

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Supplementary Fig. 1

NCM460 cells were either left untreated (“Ctrl”), or treated with applied concentrations of icaritin (ICT) for 72 h, cell proliferation was tested by MTT assay (a), and cell necrosis was tested by LDH release assay (b). Experiments in this figure were repeated three times, and similar results were obtained. For each assay, n = 5. * p < 0.05 vs. “Ctrl” group (GIF 29 kb)

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Zhou, C., Chen, Z., Lu, X. et al. Icaritin activates JNK-dependent mPTP necrosis pathway in colorectal cancer cells. Tumor Biol. 37, 3135–3144 (2016). https://doi.org/10.1007/s13277-015-4134-3

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  • DOI: https://doi.org/10.1007/s13277-015-4134-3

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