Effects of thymoquinone and etoposide combination on cell viability and genotoxicity in human cervical cancer hela cells
Year 2022,
Volume: 52 Issue: 3, 258 - 264, 30.12.2022
Hediye Gamze Nur Çelebioğlu
,
Merve Becit-kızılkaya
,
Aydan Çağlayan
,
Sevtap Aydın Dilsiz
Abstract
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 ef- fect 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. Thymo- quinone 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.
Supporting Institution
Hacettepe University Scientific Research Projects Coordination
Project Number
THD-2021-19378
References
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- Becit, M., & Aydın Dilsiz, S. (2020). An In Vitro Study on the Inter- actions of Pycnogenol® with Cisplatin in Human Cervical Cancer Cells. Turkish Journal of Pharmaceutical Sciences, 17(1), 1-6.
- Collins, A. R., Dobson, V. L., Dusinka, M., Kennedy, G., & Stetina, R. (1997). The comet assay: what can it really tell us? Mutation Re- search, 375, 183-93.
- Coutts, A. S., & La Thangue, N., (2006). The p53 response during DNA damage: impact of transcriptional cofactors. Biochemical Society Symposia, 6, 181–189.
- Dirican, A., Atmaca, H., Bozkurt, E., Erten, C., Karaca, B., & Uslu, R. (2015). Novel combination of docetaxel and thymoquinone in-duces synergistic cytotoxicity and apoptosis in DU-145 human prostate cancer cells by modulating PI3K–AKT pathway. Clinical and Translational Oncology, 17(2), 145-151.
- El-Mahdy, M. A., Zhu, Q., Wang, Q. E., Wani, G., & Wani, A. A. (2005). Thymoquinone induces apoptosis through activation of cas- pase-8 and mitochondrial events in p53-null myeloblastic leu- kemia HL-60 cells. International Journal of Cancer, 117, 409–417.
- ElKhoely, A., Hafez, H. F., Ashmawy, A. M., Badary, O., Abdelaziz, A., Mostafa, A., & Shouman, S. A. (2015). Chemopreventive and thera- peutic potentials of thymoquinone in HepG2 cells: mechanistic perspectives. Journal of Natural Medicines, 69(3), 313-23.
- 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.
- Green, J. A., Kirwan, J. M., & Tierney, J.F. (2001). Survival and re- currence after concomitant chemotherapy and radiotherapy for cancer of the uterine cervix: a systematic review and meta-analy- sis. Lancet, 358, 781–786.
- Gurung, R. L., Lim, S. N., Khaw, A. K., Soon, J.F., Shenoy, K., Mo- hamed Ali, S., … Hande, M. P. (2010). Thymoquinone induces telomere shortening, DNA damage and apoptosis in human glio- blastoma cells. PLoS One, 5(8), e12124.
- 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 Thera- py, 7, 1375-87.
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- Khader, M., Bresgen, N., & Eckl, P. M. (2009). In vitro toxicological properties of thymoquinone. Food and Chemical Toxicology, 47(1), 129-33.
- 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.
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- Kluska, M., & Woźniak, K. (2021). Natural Polyphenols as Modu- lators of Etoposide Anti-Cancer Activity. International Journal of Molecular Sciences, 20, 22(12), 6602.
- Li, F., Rajendran, P., & Sethi, G. (2010). Thymoquinone inhibits pro- liferation, induces apoptosis and chemosensitizes human multi- ple myeloma cells through suppression of signal transducer and activator of transcription 3 activation pathway. British Journal of Pharmacology, 161, 541–554.
- Mahmoud, Y. K., & Abdelrazek, H. M. A. (2019). Cancer: Thymoqui- none antioxidant/pro-oxidant effect as potential anticancer rem- edy. Biomedicine & Pharmacotherapy, 115, 108783.
- Mosmann T. (1983). Rapid colorimetric assay for cellular growth and survival: Application to proliferation and cytotoxicity assays. Journal of Immunological Method, 65, 55-63.
- Negrette-Guzmán, M. (2019). Combinations of the antioxidants sulforaphane or curcumin and the conventional antineoplastics cisplatin or doxorubicin as prospects for anticancer chemothera- py. European Journal of Pharmacology, 15(859), 172513.
- Ng, W. K., Yazan, L. S., & Ismail, M. (2011). Thymoquinone from Ni- gella sativa was more potent than cisplatin in eliminating of SiHa cells via apoptosis with down-regulation of Bcl-2 protein. Toxicol- ogy In Vitro, 25(7), 1392-8.
- Pucci, C., Martinelli, C., & Ciofani G. (2019). Innovative approaches for cancer treatment: current perspectives and new challenges. Ecancermedicalscience, 13, 961.
- Racoma, I. O., Meisen, W. H., Wang, Q. E., Kaur, B., & Wani, A. A. (2013). Thymoquinone inhibits autophagy and induces cathep- sin-mediated, caspase-independent cell death in glioblastoma cells. PLoS One, 8(9), e72882.
- 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.
- 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.
- 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.
- 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.
- 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 Oncol- ogy, 22, 107–113.
- 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.
- Sinkule, J. A. (1984). Etoposide: a semisynthetic epipodophyllo- toxin. Chemistry, pharmacology, pharmacokinetics, adverse ef- fects and use as an antineoplastic agent. Pharmacotherapy, 4(2), 61-73.
- 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 Jour- nal for Clinicians, 71(3), 209-249.
- Surendiran, A., Balamurugan, N., Gunaseelan, K., Akhtar, S., Reddy, K. S., & Adithan, C. (2010). Adverse drug reaction profile of cispl- atin-based chemotherapy regimen in a tertiary care hospital in India: An evaluative study. Indian Journal of Pharmacology, 42(1), 40-3.
- Woo, C. C., Kumar, A. P., Sethi, G., & Tan, K. H. (2012). Thymoqui- none: potential cure for inflammatory disorders and cancer. Bio- chemical Pharmacology, 15, 83(4), 443-51.
- 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 Me- dicinal Chemistry, 10, 80, 267-77.
- 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 signal- ing pathway. Biochemical and Biophysical Research Communica- tions, 19, 545, 81-88.
- Yıldırım, H. Y., Azzawri, A. A., & Duran, T. (2019). Thymoquinone in- duces apoptosis via targeting the Bax/BAD and Bcl-2 pathway in breast cancer cells. Dicle Tıp Dergisi, 46(3), 411-417.
Year 2022,
Volume: 52 Issue: 3, 258 - 264, 30.12.2022
Hediye Gamze Nur Çelebioğlu
,
Merve Becit-kızılkaya
,
Aydan Çağlayan
,
Sevtap Aydın Dilsiz
Project Number
THD-2021-19378
References
- Al-Shdefat, R. I., Abd-ElAziz, M. A., & Al-Saikhan, F. I. (2014). Geno- protective and Genotoxic Effects of Thymoquinone on Doxoru- bicin-Induced Damage in Isolated Human Leukocytes. Tropical Journal of Pharmaceutical Research, 13 (12), 2015-2020.
- Ashley, R. E. & Osheroff, N. (2014). Natural products as topoisom- erase II poisons: effects of thymoquinone on DNA cleavage medi- ated by human topoisomerase IIα. Chemical Research in Toxicol- ogy, 27(5), 787-93.
- 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 Pharma- cology, 27(5), 557-69.
- 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.
- Becit, M., & Aydın Dilsiz, S. (2020). An In Vitro Study on the Inter- actions of Pycnogenol® with Cisplatin in Human Cervical Cancer Cells. Turkish Journal of Pharmaceutical Sciences, 17(1), 1-6.
- Collins, A. R., Dobson, V. L., Dusinka, M., Kennedy, G., & Stetina, R. (1997). The comet assay: what can it really tell us? Mutation Re- search, 375, 183-93.
- Coutts, A. S., & La Thangue, N., (2006). The p53 response during DNA damage: impact of transcriptional cofactors. Biochemical Society Symposia, 6, 181–189.
- Dirican, A., Atmaca, H., Bozkurt, E., Erten, C., Karaca, B., & Uslu, R. (2015). Novel combination of docetaxel and thymoquinone in-duces synergistic cytotoxicity and apoptosis in DU-145 human prostate cancer cells by modulating PI3K–AKT pathway. Clinical and Translational Oncology, 17(2), 145-151.
- El-Mahdy, M. A., Zhu, Q., Wang, Q. E., Wani, G., & Wani, A. A. (2005). Thymoquinone induces apoptosis through activation of cas- pase-8 and mitochondrial events in p53-null myeloblastic leu- kemia HL-60 cells. International Journal of Cancer, 117, 409–417.
- ElKhoely, A., Hafez, H. F., Ashmawy, A. M., Badary, O., Abdelaziz, A., Mostafa, A., & Shouman, S. A. (2015). Chemopreventive and thera- peutic potentials of thymoquinone in HepG2 cells: mechanistic perspectives. Journal of Natural Medicines, 69(3), 313-23.
- 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.
- Green, J. A., Kirwan, J. M., & Tierney, J.F. (2001). Survival and re- currence after concomitant chemotherapy and radiotherapy for cancer of the uterine cervix: a systematic review and meta-analy- sis. Lancet, 358, 781–786.
- Gurung, R. L., Lim, S. N., Khaw, A. K., Soon, J.F., Shenoy, K., Mo- hamed Ali, S., … Hande, M. P. (2010). Thymoquinone induces telomere shortening, DNA damage and apoptosis in human glio- blastoma cells. PLoS One, 5(8), e12124.
- 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 Thera- py, 7, 1375-87.
- Hansen, M. B., Nielsen, S. E., & Berg, K. (1989). Re-examination and further development of a precise and rapid dye method for mea- suring cell growth/cell kill. Journal of Immunological Methods, 119, 203-210.
- Khader, M., Bresgen, N., & Eckl, P. M. (2009). In vitro toxicological properties of thymoquinone. Food and Chemical Toxicology, 47(1), 129-33.
- 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.
- Khan, M. A., Tania, M., & Fu, J. (2019). Epigenetic role of thymo- quinone: impact on cellular mechanism and cancer therapeu- tics. Drug Discovery Today, 24(12), 2315-2322.
- Kluska, M., & Woźniak, K. (2021). Natural Polyphenols as Modu- lators of Etoposide Anti-Cancer Activity. International Journal of Molecular Sciences, 20, 22(12), 6602.
- Li, F., Rajendran, P., & Sethi, G. (2010). Thymoquinone inhibits pro- liferation, induces apoptosis and chemosensitizes human multi- ple myeloma cells through suppression of signal transducer and activator of transcription 3 activation pathway. British Journal of Pharmacology, 161, 541–554.
- Mahmoud, Y. K., & Abdelrazek, H. M. A. (2019). Cancer: Thymoqui- none antioxidant/pro-oxidant effect as potential anticancer rem- edy. Biomedicine & Pharmacotherapy, 115, 108783.
- Mosmann T. (1983). Rapid colorimetric assay for cellular growth and survival: Application to proliferation and cytotoxicity assays. Journal of Immunological Method, 65, 55-63.
- Negrette-Guzmán, M. (2019). Combinations of the antioxidants sulforaphane or curcumin and the conventional antineoplastics cisplatin or doxorubicin as prospects for anticancer chemothera- py. European Journal of Pharmacology, 15(859), 172513.
- Ng, W. K., Yazan, L. S., & Ismail, M. (2011). Thymoquinone from Ni- gella sativa was more potent than cisplatin in eliminating of SiHa cells via apoptosis with down-regulation of Bcl-2 protein. Toxicol- ogy In Vitro, 25(7), 1392-8.
- Pucci, C., Martinelli, C., & Ciofani G. (2019). Innovative approaches for cancer treatment: current perspectives and new challenges. Ecancermedicalscience, 13, 961.
- Racoma, I. O., Meisen, W. H., Wang, Q. E., Kaur, B., & Wani, A. A. (2013). Thymoquinone inhibits autophagy and induces cathep- sin-mediated, caspase-independent cell death in glioblastoma cells. PLoS One, 8(9), e72882.
- 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.
- 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.
- 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.
- 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.
- 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 Oncol- ogy, 22, 107–113.
- 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.
- Sinkule, J. A. (1984). Etoposide: a semisynthetic epipodophyllo- toxin. Chemistry, pharmacology, pharmacokinetics, adverse ef- fects and use as an antineoplastic agent. Pharmacotherapy, 4(2), 61-73.
- 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 Jour- nal for Clinicians, 71(3), 209-249.
- Surendiran, A., Balamurugan, N., Gunaseelan, K., Akhtar, S., Reddy, K. S., & Adithan, C. (2010). Adverse drug reaction profile of cispl- atin-based chemotherapy regimen in a tertiary care hospital in India: An evaluative study. Indian Journal of Pharmacology, 42(1), 40-3.
- Woo, C. C., Kumar, A. P., Sethi, G., & Tan, K. H. (2012). Thymoqui- none: potential cure for inflammatory disorders and cancer. Bio- chemical Pharmacology, 15, 83(4), 443-51.
- 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 Me- dicinal Chemistry, 10, 80, 267-77.
- 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 signal- ing pathway. Biochemical and Biophysical Research Communica- tions, 19, 545, 81-88.
- Yıldırım, H. Y., Azzawri, A. A., & Duran, T. (2019). Thymoquinone in- duces apoptosis via targeting the Bax/BAD and Bcl-2 pathway in breast cancer cells. Dicle Tıp Dergisi, 46(3), 411-417.