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Reactive oxygen species mediate thymoquinone-induced apoptosis and activate ERK and JNK signaling

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Abstract

Thymoquinone (TQ), a component of black seed essential oil, is known to induce apoptotic cell death and oxidative stress, however, the direct involvement of oxidants in TQ-induced cell death has not been established yet. Here, we show that TQ inhibited the proliferation of a panel of human colon cancer cells (Caco-2, HCT-116, LoVo, DLD-1 and HT-29), without exhibiting cytotoxicity to normal human intestinal FHs74Int cells. Further investigation in DLD-1 revealed that apoptotic cell death is the mechanism for TQ-induced growth inhibition as confirmed by flow cytometry, M30 cytodeath and caspase-3/7 activation. Apoptosis was induced via the generation of reactive oxygen species (ROS) as evidenced by the abrogation of TQ apoptotic effect in cells preincubated with the strong antioxidant N-acetyl cysteine (NAC). TQ increased the phosphorylation states of the mitogen-activated protein kinases (MAPK) JNK and ERK, but not of p38. Their activation was completely abolished in the presence of NAC. Using PD98059 and SP600125, specific ERK and JNK inhibitors, the two kinases were found to possess pro-survival activities in TQ-induced cell death. These data present evidence linking the pro-oxidant effects of TQ with its apoptotic effects in colon cancer and prove a protective role of MAPK.

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Acknowledgments

We thank members of the Central Research Science Laboratory at the American University of Beirut, Lebanon for their help in using the flow cytometer and HPLC. We thank Isabel Zeitträger and Astrid Taut from Department of Medicine 1, Erlangen, Germany for their technical assistance. This work was supported by Deutsche Forschungsgemeinschaft (SCHN477/7-3, SCHN477/7-4) and by the University Research Board of the AUB and the Lebanese National Council for Scientific Research. Nahed El-Najjar was partly supported by the DAAD.

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Authors and Affiliations

  1. Department of Biology, American University of Beirut, Beirut, Lebanon

    Nahed El-Najjar, Manal Chatila, Hiba Moukadem & Hala Gali-Muhtasib

  2. Division of Pharmaceutical Biology, Faculty of Pharmacy, University of Helsinki, Helsinki, Finland

    Nahed El-Najjar & Heikki Vuorela

  3. Institute for Surgical Research, Philipps University Marburg, Marburg, Germany

    Matthias Ocker

  4. Experimental Tumorpathology, Institute for Pathology, University Erlangen-Nuremberg, Erlangen, Germany

    Muktheshwar Gandesiri & Regine Schneider-Stock

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  1. Nahed El-Najjar
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Correspondence to Hala Gali-Muhtasib.

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El-Najjar, N., Chatila, M., Moukadem, H. et al. Reactive oxygen species mediate thymoquinone-induced apoptosis and activate ERK and JNK signaling. Apoptosis 15, 183–195 (2010). https://doi.org/10.1007/s10495-009-0421-z

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