Research Article


DOI :10.26650/IstanbulJPharm.2022.1105443   IUP :10.26650/IstanbulJPharm.2022.1105443    Full Text (PDF)

Effects of thymoquinone and etoposide combination on cell viability and genotoxicity in human cervical cancer hela cells

Hediye Gamze Nur ÇelebioğluMerve BecitAydan ÇağlayanSevtap Aydın Dilsiz

Background and Aims: It is thought that thymoquinone might have a crucial role in preventing DNA damage, regulating DNA repair mechanisms, and inhibiting the formation of a cancer. Studies on the cytotoxic and genotoxic effects of thymoquinone together with etoposide in cervical carcinoma cells (HeLa) are not adequate. The objective of this study is to evaluate the effect of combinations with thymoquinone on etoposide cytotoxicity and genotoxicity in HeLa cells. Methods: Cytotoxicity was evaluated by MTT assay and genotoxicity was determined by Comet assay. Results: The IC50 values of thymoquinone were 233.6 μM and 145.5 μM, and the IC50 values of etoposide were 167.3 μM and 52.7 μM for 24 and 48 h, respectively. Thymoquinone significantly decreased the approximate IC50 value of etoposide in doses of 15.63 μM and above for 24 h and 31.5 μM and above for 48 h in a dose-dependent manner. 0.1-5 μM thymoquinone and 1 μM etoposide alone did not cause DNA damage, but at higher doses increased DNA damage significantly in a dose-dependent manner. Thymoquinone significantly reduced DNA damage induced by 10 μM etoposide at the doses of 0.1-10 μM. Conclusion: Our results show that thymoquinone might increase the cytotoxic and genotoxic effects of etoposide in HeLa cells at high doses and reduce DNA damage at low doses that are not cytotoxic, which suggests that etoposide may increase its anticancer effect at high doses, but comprehensive studies are needed on this subject. This study is a preliminary study and will contribute to the development of new treatment strategies.


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References

  • Al-Shdefat, R. I., Abd-ElAziz, M. A., & Al-Saikhan, F. I. (2014). Geno-protective and Genotoxic Effects of Thymoquinone on Doxorubicin-Induced Damage in Isolated Human Leukocytes. Tropical Journal of Pharmaceutical Research, 13 (12), 2015-2020. google scholar
  • Ashley, R. E. & Osheroff, N. (2014). Natural products as topoisomerase II poisons: effects of thymoquinone on DNA cleavage mediated by human topoisomerase Ila. Chemical Research in Toxicology, 27(5), 787-93. google scholar
  • Attoub, S., Sperandio, O., Raza, H., Arafat, K., Al-Salam, S., Al Sultan, M. A., ..., Adem, A. (2013). Thymoquinone as an anticancer agent: evidence from inhibition of cancer cells viability and invasion in vitro and tumor growth in vivo. Fundamental & Clinical Pharmacology, 27(5), 557-69. google scholar
  • Bashmail, H. A., Aliaa, A.A., Abdulwahab, N., Gehan, A. H., Ghada, A., Hani, C., & Ahmed, M. A. (2018). Thymoquinone synergizes gemcitabine anti-breast cancer activity via modulating its apop-totic and autophagic activities. Scientific Reports, 8, 11674. google scholar
  • Becit, M., & Aydın Dilsiz, S. (2020). An In Vitro Study on the Interactions of Pycnogenol® with Cisplatin in Human Cervical Cancer Cells. Turkish Journal of Pharmaceutical Sciences, 17(1), 1-6. google scholar
  • Collins, A. R., Dobson, V. L., Dusinka, M., Kennedy, G., & Stetina, R. (1997). The comet assay: what can it really tell us? Mutation Research, 375, 183-93. google scholar
  • Coutts, A. S., & La Thangue, N., (2006). The p53 response during DNA damage: impact of transcriptional cofactors. Biochemical Society Symposia, 6, 181-189. google scholar
  • Dirican, A., Atmaca, H., Bozkurt, E., Erten, C., Karaca, B., & Uslu, R. (2015). Novel combination of docetaxel and thymoquinone induces synergistic cytotoxicity and apoptosis in DU-145 human prostate cancer cells by modulating PI3K-AKT pathway. Clinical and Translational Oncology, 17(2), 145-151. google scholar
  • El-Mahdy, M. A., Zhu, Q., Wang, Q. E., Wani, G., & Wani, A. A. (2005). Thymoquinone induces apoptosis through activation of caspase-8 and mitochondrial events in p53-null myeloblastic leukemia HL-60 cells. International Journal of Cancer, 117, 409-417. google scholar
  • ElKhoely, A., Hafez, H. F., Ashmawy, A. M., Badary, O., Abdelaziz, A., Mostafa, A., & Shouman, S. A. (2015). Chemopreventive and therapeutic potentials of thymoquinone in HepG2 cells: mechanistic perspectives. Journal of Natural Medicines, 69(3), 313-23. google scholar
  • Gali-Muhtasib, H. U., Abou Kheir, W. G., Kheir, L. A., Darwiche, N., & Crooks, P. A. (2004). Molecular pathway for thymoquinone-induced cell-cycle arrest and apoptosis in neoplastic keratino-cytes. Anti-cancer Drugs, 15(4), 389-399. google scholar
  • Green, J. A., Kirwan, J. M., & Tierney, J.F. (2001). Survival and recurrence after concomitant chemotherapy and radiotherapy for cancer of the uterine cervix: a systematic review and meta-analysis. Lancet, 358, 781-786. google scholar
  • Gurung, R. L., Lim, S. N., Khaw, A. K., Soon, J.F., Shenoy, K., Mohamed Ali, S., ... Hande, M. P (2010). Thymoquinone induces telomere shortening, DNA damage and apoptosis in human glioblastoma cells. PLoS One, 5(8), e12124. google scholar
  • Hafiza, W. A., & Latifah, S. Y. (2014). Potential implications of GRP58 expression and susceptibility of cervical cancer to cisplatin and thymoquinone-based therapy. Journal of OncoTargets and Therapy, 7, 1375-87. google scholar
  • Hansen, M. B., Nielsen, S. E., & Berg, K. (1989). Re-examination and further development of a precise and rapid dye method for measuring cell growth/cell kill. Journal of Immunological Methods, 119, 203-210. google scholar
  • Khader, M., Bresgen, N., & Eckl, P. M. (2009). In vitro toxicological properties of thymoquinone. Food and Chemical Toxicology, 47(1), 129-33. google scholar
  • Khalife, R., Hodroj, M. H., Fakhoury, R., & Rizk, S. (2016). Thymoqui-none from Nigella sativa seeds promotes the antitumor activity of noncytotoxic doses of topotecan in human colorectal cancer cells in vitro. Planta Medica, 82(4), 312-21. google scholar
  • Khan, M. A., Tania, M., & Fu, J. (2019). Epigenetic role of thymo-quinone: impact on cellular mechanism and cancer therapeutics. Drug Discovery Today, 24(12), 2315-2322. google scholar
  • Kluska, M., & Wozniak, K. (2021). Natural Polyphenols as Modulators of Etoposide Anti-Cancer Activity. International Journal of Molecular Sciences, 20, 22(12), 6602. google scholar
  • Li, F., Rajendran, P., & Sethi, G. (2010). Thymoquinone inhibits proliferation, induces apoptosis and chemosensitizes human multiple myeloma cells through suppression of signal transducer and activator of transcription 3 activation pathway. British Journal of Pharmacology, 161, 541-554. google scholar
  • Mahmoud, Y. K., & Abdelrazek, H. M. A. (2019). Cancer: Thymoqui-none antioxidant/pro-oxidant effect as potential anticancer remedy. Biomedicine & Pharmacotherapy, 115, 108783. google scholar
  • Mosmann T. (1983). Rapid colorimetric assay for cellular growth and survival: Application to proliferation and cytotoxicity assays. Journal of Immunological Method, 65, 55-63. google scholar
  • Negrette-Guzman, M. (2019). Combinations of the antioxidants sulforaphane or curcumin and the conventional antineoplastics cisplatin or doxorubicin as prospects for anticancer chemotherapy. European Journal of Pharmacology, 15(859), 172513. google scholar
  • Ng, W. K., Yazan, L. S., & Ismail, M. (2011). Thymoquinone from Nigella sativa was more potent than cisplatin in eliminating of SiHa cells via apoptosis with down-regulation of Bcl-2 protein. Toxicology In Vitro, 25(7), 1392-8. google scholar
  • Pucci, C., Martinelli, C., & Ciofani G. (2019). Innovative approaches for cancer treatment: current perspectives and new challenges. Ecancermedicalscience, 13, 961. google scholar
  • Racoma, I. O., Meisen, W. H., Wang, Q. E., Kaur, B., & Wani, A. A. (2013). Thymoquinone inhibits autophagy and induces cathepsin-mediated, caspase-independent cell death in glioblastoma cells. PLoS One, 8(9), e72882. google scholar
  • Rahmani, A. H., Alzohairy, M. A., Khan, M.A., & Aly, S. M. (2014). Therapeutic implications of black seed and its constituent thy-moquinone in the prevention of cancer through inactivation and activation of molecular pathways. Evidence-Based Complementary and Alternative Medicine, 724658. google scholar
  • Rello-Varonai, S., Gámez, A., Moreno, V., Stockert, J.C., Cristóbal, J., Pacheco, M.,... Villanueva, A., (2006). Metaphase arrest and cell death induced by etoposide on HeLa cells. International Journal of Biochemistry & Cell Biology, 38(12), 2183-95. google scholar
  • Sakalar, C., Yuruk, M., Kaya, T., Aytekin, M., Kuk, S., & Canatan, H. (2013). Pronounced transcriptional regulation of apoptotic and TNF-NF-kappa-B signaling genes during the course of thymo-quinone mediated apoptosis in HeLa cells. Molecular and Cellular Biochemistry, 383(1-2), 243-51. google scholar
  • Salvo, G., Gonzalez Martin, A., Gonzales, N. R., & Frumovitz, M. (2019). Updates and management algorithm for neuroendocrine tumors of the uterine cervix. International Journal of Gynecological Cancer, 29(6), 986-995. google scholar
  • Shoieb, A. M., Elgayyar, M., Dudrick, P. S., Bell, J. L., & Tithof, P. K. (2003). In vitro inhibition of growth and induction of apoptosis in cancer cell lines by thymoquinone. International Journal of Oncology, 22, 107-113. google scholar
  • Singh, N. P., McCoy, M. T., Tice, R. R., & Schneider, E. L. (1988). A simple technique for quantitation of low levels of DNA damage in individual cells. Experimental Cell Research, 175, 184-91. google scholar
  • Sinkule, J. A. (1984). Etoposide: a semisynthetic epipodophyllo-toxin. Chemistry, pharmacology, pharmacokinetics, adverse effects and use as an antineoplastic agent. Pharmacotherapy, 4(2), 61-73. google scholar
  • Sung, H., Ferlay, J., Siegel, R.L., Laversanne, M., Soerjomataram, I., Jemal, A., & Bray, F. (2021). GLOBOCAN Estimates of Incidence and Mortality Worldwide for 36 Cancers in 185 Countries. Cancer Journal for Clinicians, 71(3), 209-249. google scholar
  • Surendiran, A., Balamurugan, N., Gunaseelan, K., Akhtar, S., Reddy, K. S., & Adithan, C. (2010). Adverse drug reaction profile of cisplatin-based chemotherapy regimen in a tertiary care hospital in India: An evaluative study. Indian Journal of Pharmacology, 42(1), 40-3. google scholar
  • Woo, C. C., Kumar, A. P., Sethi, G., & Tan, K. H. (2012). Thymoqui-none: potential cure for inflammatory disorders and cancer. Biochemical Pharmacology, 15, 83(4), 443-51. google scholar
  • Xiao, L., Zhao, W., Li, H. M., Wan, D. J., Li, D.S., Chen, T., & Tang, Y. J. (2014). Design and synthesis of the novel DNA topoisomerase II inhibitors: Esterification and amination substituted 4’-demethy-lepipodophyllotoxin derivates exhibiting anti-tumor activity by activating ATM/ATR signaling pathways. European Journal of Medicinal Chemistry, 10, 80, 267-77. google scholar
  • Xiaofei, J., Mingqing, S., Miao, S., Yizhen, Y., Shuang, Z., Qinhua, X., & Kai, Z. (2021). Oleanolic acid inhibits cervical cancer Hela cell proliferation through modulation of the ACSL4 ferroptosis signaling pathway. Biochemical and Biophysical Research Communications, 19, 545, 81-88. google scholar
  • Yıldırım, H. Y., Azzawri, A. A., & Duran, T. (2019). Thymoquinone induces apoptosis via targeting the Bax/BAD and Bcl-2 pathway in breast cancer cells. Dicle Tıp Dergisi, 46(3), 411-417. google scholar

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APA

Nur Çelebioğlu, H.G., Becit, M., Çağlayan, A., & Aydın Dilsiz, S. (2022). Effects of thymoquinone and etoposide combination on cell viability and genotoxicity in human cervical cancer hela cells. İstanbul Journal of Pharmacy, 52(3), 258-264. https://doi.org/10.26650/IstanbulJPharm.2022.1105443


AMA

Nur Çelebioğlu H G, Becit M, Çağlayan A, Aydın Dilsiz S. Effects of thymoquinone and etoposide combination on cell viability and genotoxicity in human cervical cancer hela cells. İstanbul Journal of Pharmacy. 2022;52(3):258-264. https://doi.org/10.26650/IstanbulJPharm.2022.1105443


ABNT

Nur Çelebioğlu, H.G.; Becit, M.; Çağlayan, A.; Aydın Dilsiz, S. Effects of thymoquinone and etoposide combination on cell viability and genotoxicity in human cervical cancer hela cells. İstanbul Journal of Pharmacy, [Publisher Location], v. 52, n. 3, p. 258-264, 2022.


Chicago: Author-Date Style

Nur Çelebioğlu, Hediye Gamze, and Merve Becit and Aydan Çağlayan and Sevtap Aydın Dilsiz. 2022. “Effects of thymoquinone and etoposide combination on cell viability and genotoxicity in human cervical cancer hela cells.” İstanbul Journal of Pharmacy 52, no. 3: 258-264. https://doi.org/10.26650/IstanbulJPharm.2022.1105443


Chicago: Humanities Style

Nur Çelebioğlu, Hediye Gamze, and Merve Becit and Aydan Çağlayan and Sevtap Aydın Dilsiz. Effects of thymoquinone and etoposide combination on cell viability and genotoxicity in human cervical cancer hela cells.” İstanbul Journal of Pharmacy 52, no. 3 (Feb. 2023): 258-264. https://doi.org/10.26650/IstanbulJPharm.2022.1105443


Harvard: Australian Style

Nur Çelebioğlu, HG & Becit, M & Çağlayan, A & Aydın Dilsiz, S 2022, 'Effects of thymoquinone and etoposide combination on cell viability and genotoxicity in human cervical cancer hela cells', İstanbul Journal of Pharmacy, vol. 52, no. 3, pp. 258-264, viewed 1 Feb. 2023, https://doi.org/10.26650/IstanbulJPharm.2022.1105443


Harvard: Author-Date Style

Nur Çelebioğlu, H.G. and Becit, M. and Çağlayan, A. and Aydın Dilsiz, S. (2022) ‘Effects of thymoquinone and etoposide combination on cell viability and genotoxicity in human cervical cancer hela cells’, İstanbul Journal of Pharmacy, 52(3), pp. 258-264. https://doi.org/10.26650/IstanbulJPharm.2022.1105443 (1 Feb. 2023).


MLA

Nur Çelebioğlu, Hediye Gamze, and Merve Becit and Aydan Çağlayan and Sevtap Aydın Dilsiz. Effects of thymoquinone and etoposide combination on cell viability and genotoxicity in human cervical cancer hela cells.” İstanbul Journal of Pharmacy, vol. 52, no. 3, 2022, pp. 258-264. [Database Container], https://doi.org/10.26650/IstanbulJPharm.2022.1105443


Vancouver

Nur Çelebioğlu HG, Becit M, Çağlayan A, Aydın Dilsiz S. Effects of thymoquinone and etoposide combination on cell viability and genotoxicity in human cervical cancer hela cells. İstanbul Journal of Pharmacy [Internet]. 1 Feb. 2023 [cited 1 Feb. 2023];52(3):258-264. Available from: https://doi.org/10.26650/IstanbulJPharm.2022.1105443 doi: 10.26650/IstanbulJPharm.2022.1105443


ISNAD

Nur Çelebioğlu, HediyeGamze - Becit, Merve - Çağlayan, Aydan - Aydın Dilsiz, Sevtap. Effects of thymoquinone and etoposide combination on cell viability and genotoxicity in human cervical cancer hela cells”. İstanbul Journal of Pharmacy 52/3 (Feb. 2023): 258-264. https://doi.org/10.26650/IstanbulJPharm.2022.1105443



TIMELINE


Submitted20.04.2022
Accepted15.08.2022
Published Online30.12.2022

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