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Pronounced transcriptional regulation of apoptotic and TNF–NF-kappa-B signaling genes during the course of thymoquinone mediated apoptosis in HeLa cells

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Abstract

Thymoquinone (TQ) is the active ingredient extracted from the essential oil of Nigella sativa. A number of studies implicated TQ as an antitumor agent. In this study, cytotoxic effects of the oil of N. sativa and TQ were evaluated on human cervical cancer cell line, HeLa cells. IC50 value was ~0.125 μl/ml for N. sativa oil preparations and 12.5 μM for TQ. TQ strongly inhibited wound healing at all concentrations ranging from 12.5 to 100 μM in a scratch wound healing assay. Additionally, induction of apoptosis by TQ was assessed by Giemsa staining and TQ was found to induce apoptosis in cancer cells especially at concentrations of 50 and 100 μM. TQ-mediated transcriptional regulation of 84 genes involved in apoptosis was studied using a PCR array. At low dose (12.5 μM), TQ was found to induce expression of four pro-apoptotic genes: BIK (~22.7-fold), FASL (~2.9-fold), BCL2L10 (~2.1-fold), and CASP1 (~2-fold). TQ was also found to reduce the expression of an anti-apoptotic gene implicated in NF-kappa-B signaling and cancer: RELA (~8-fold). At high dose (100 μM), TQ mediated the expression of 21 genes implicated directly in apoptosis (6 genes), TNF signaling (10 genes), and NF-kappa-B signaling (3 genes) such as BIK, BID, TNFRSF10A, TNFRSF10B, TNF, TRAF3, RELA, and RELB. In conclusion, this study implicates the role of TQ in the inhibition of cancer cell proliferation and migration. At the same time, our results strongly suggest that TQ intervenes with TNF and NF-kappa-B signaling during TQ-mediated induction of apoptosis in cancer cells.

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Acknowledgments

We thank Dr. Nazmiye Bitgen for her technical support in Giemsa staining of cancer cells and assessment of apoptotic cells. This work was supported by Erciyes University Scientific Research Fund (EU-BAP), Grant Number: CAP-12-4044 (C. SAKALAR).

Conflict of Interest

All of the authors declare that they have no conflict of interest and no financial relationship with any commercial entity that has an interest in the subject of this manuscript.

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Authors and Affiliations

  1. Department of Medical Biology, Faculty of Medicine, Erciyes University, Melikgazi, 38039, Kayseri, Turkey

    Cagri Sakalar, Metin Aytekin & Halit Canatan

  2. Genome and Stem Cell Research Center, Erciyes University, Melikgazi, 38039, Kayseri, Turkey

    Cagri Sakalar, Metin Aytekin, Salih Kuk & Halit Canatan

  3. Department of Parasitology, Faculty of Medicine, Erciyes University, Melikgazi, 38039, Kayseri, Turkey

    Merve Yuruk, Tugba Kaya & Salih Kuk

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  1. Cagri Sakalar
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  2. Merve Yuruk
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Correspondence to Cagri Sakalar.

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Sakalar, C., Yuruk, M., Kaya, T. et al. Pronounced transcriptional regulation of apoptotic and TNF–NF-kappa-B signaling genes during the course of thymoquinone mediated apoptosis in HeLa cells. Mol Cell Biochem 383, 243–251 (2013). https://doi.org/10.1007/s11010-013-1772-x

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