ORIGINAL RESEARCH

The Effects of Thymoquinone, A Bioactive Compound of Nigella Sativa, in Combination With Cisplatin on the Viability of HeLa Cervical Cancer Cells
Received Date : 10 Sep 2021
Accepted Date : 16 Dec 2021
Available Online : 23 May 2022
Doi: 10.56054/ajohoim.2021-86165 - Article's Language: EN
Acad. J. Homeopat. & Integ. Med. 2023;1(1):1-10
ABSTRACT
Objective: Thymoquinone, a monoterpene molecule which is derived from the volatile oil of Nigella sativa L. seed known as black seed “çörek otu” in Turkey, is a bioactive compound. Although it is consumed as food, it is claimed that thymoquinone has a wide range of effects, from its anticancer effect to its antiallergic effect, together with its main antioxidant property. Thymoquinone is thought to play an important role in the prevention and treatment of various types of cancers. Cisplatin, a platinum-based anticancer agent, has been used for many years in the treatment of solid tumors such as ovarian, cervix, testis, prostate, bladder and lung cancers despite its toxic effects. The use of plant extracts in combination with chemotherapeutic drugs in order to reduce the side effects of drugs used in chemotherapy and to increase their intra-body effects is the focus of research. The aim of this study is to reveal the cytotoxic effects of the combined use of thymoquinone, which is known to have important biological effects and whose importance has increased in recent studies, together with the anticancer drug cisplatin, on cervical cancer cells (HeLa). Material and Methods: HeLa cervical carcinoma cells provided from the American Type Culture Collection were used. The effects of thymoquinone in the concentration range of 1.95 μM-1000 μM and cisplatin in the concentration range of 0.49 μM - 250 μM on HeLa cell viability were measured by MTT method after 24 and 48 hours of incubation. After determining the IC50 values of each substance alone, the effects of combinations of thymoquinone with IC50 doses of cisplatin on cell viability were determined by MTT method after 24 and 48 hours of incubation. Results: In HeLa cells, for 24 h and 48 h incubations, IC50 values of thymoquinone were found to be 143.7 μM and 67.5 μM, respectively and IC50 values of cisplatin were found to be 20.3 μM and 12.9 μM, respectively. According to the study on the effects of the combined administration of thymoquinone on cisplatin cytotoxicity. Thymoquinone in the concentration range of 7.8-250 μM for 24 hours of incubation, statistically decreased the IC50 value of cisplatin (20 μM) in a dose-dependent manner (20.6%, 33.3%, 46.8%, 56.5%, 70.8%, 84.2% for 7.8 μM, 15.6 μM, 31.3 μM, 62.5 μM, 125 μM, 250 μM, respectively) in HeLa cells. Thymoquinone in the concentration range of 15.68-250 μM for 48 hours of incubation, statistically decreased the IC50 value of cisplatin in a dose-dependent manner (41.6%, 44.2%, 62.2%, 71.1%, 81.9% for 15.6 μM, 31.3 μM, 62.5 μM, 125 μM, 250 μM, respectively). Conclusion: In conclusion, our findings show that the combination of thymoquinone with cisplatin can increase the cytotoxicity of cisplatin in HeLa cells, and therefore it is thought that thymoquinone may increase the anticancer effect of cisplatin; however, to confirm their clinical use and to determine its interactions with anticancer drugs, the advanced in vitro and in vivo studies are needed.
REFERENCES
  1. Gullett NP, Ruhul Amin AR, Bayraktar S, Pezzuto JM, Shin DM, Khuri FR, et al. Cancer prevention with natural compounds. Semin Oncol. 2010;37:258-81. [Crossref]  [PubMed] 
  2. AbuKhader MM. Thmoquinone in the clinical treatment of cancer: fact or fiction? Pharmacogn Rev. 2013;7(14):117-20. [Crossref]  [PubMed]  [PMC] 
  3. Ng WK, Yazan LS, Yap LH, Hafiza WA ve ark. Thymoquinone-loaded nanostructured lipid carrier exhibited cytotoxicity towards breast canser cell lines (MDA-MB-231 and MCF-7) and cervical cancer cell lines (HeLa and SiHa). Biomed Res Int. 2015;2015:263131. [Crossref]  [PubMed]  [PMC] 
  4. Mostafa, AGM Hossain MK, Basak, D Bin Sayeed MS. Thymoquinone as potential adjuvant therapy for cancer treatment: evidence from preclinical studies. Front Pharmacol. 2017;8:295. [Crossref]  [PubMed]  [PMC] 
  5. Majdalawieh AF, Fayyad MW, Nasrallah GK. Anti-cancer properties and mechanisms of action of thymoquinone, the major active ingredient of Nigella sativa. Crit Rev Food Sci Nutr. 2017;57(18):3911-28. [Crossref]  [PubMed] 
  6. Ramadan MF. Nutritional value, functional properties and nutraceutical application of black cumin (Nigella sativa L.): an overview. IJFS Technol. 2007;42:1208-18. [Crossref] 
  7. Ragheb A, Attia A, Eldin WS, Elbarbry F, Gazarin S, Shoker A. The protective effect of thymoquinone, an anti-oxidant and anti-inflammatory agent, against renal injury: a review. Saaudi J Kidney Dis Transpl. 2009;20(5):741-52.
  8. Cetin R, Devrim E, Kiliçoğlu B, Avci A, Candir O, Durak I. Cisplatin impairs antioxidant system and causes oxidation in rat kidney tissue: possible portective roles of natural antioxidant foods. J Appl Toxicol. 2006;26(1):42-6. [Crossref]  [PubMed] 
  9. Dasari S, Tchounwou PB. Cisplatin in cancer therapy: molecular mechanisms of action. Eur J Pharmacol. 2014;740:364-78. [Crossref]  [PubMed]  [PMC] 
  10. Becit M, Aydin Dilsiz S, Başaran N. Effects of pycnogenol and its combinations with cisplatin on hepatocellular carcinoma cell viability. FABAD J Pharm Sci. 2021;46(1):13-22.
  11. Khader M, Eckl PM. Thymoquinone: an emerging natural drug with a wide range of medical applications. Iran J Basic Med Sci. 2014;17(12): 950-7.
  12. Darakhshan S, Bidmeshki PA, Hosseinzadeh CA, Sisakhtnezhad S. Thymoquinone and its therapeutic potentials. Pharmacological Res. 2015;95-96:138-58. [Crossref]  [PubMed] 
  13. Tchounwou PB, Dasari S, Noubissi FK, Ray P, Kumar S. Advances in our understanding of the molecular mechanisms of action of cisplatin in cancer therapy. J Exp Pharmacol. 2021;13:303-28. [Crossref]  [PubMed]  [PMC] 
  14. Chan MM, Fong D. Overcoming ovarian cancer drug resistance with phytochemicals and other compounds. In: Drug Resistance Neoplasms. Varie EG (ed.). New York, Nova Science Publishers. 2007.
  15. Sarkar FH, Li Y. Using chemopreventive agents to enhance the efficacy of cancer therapy. Cancer Res. 2006;66(7):3347-50. [Crossref]  [PubMed] 
  16. Hafiza WA, Latifah SY. Potential implications of GRP58 expression and susceptibility of cervical cancer to cisplatin and thymoquinone-based therapy. Onco. Targets. Ther. 2014;7:1375-87. [Crossref]  [PubMed]  [PMC] 
  17. Khader M, Bresgen N, Eckl PM. In vitro toxicological properties of thymoquinone. Food Chem. Toxicol. 2009;.47(1):129-33. [Crossref]  [PubMed] 
  18. Peng L, Liu A, Shen Y, Xu HZ, Yang SZ, Ying XZ, et al. Antitumor and anti-angiogenesis effects of thymoquinone on osteosarcoma through the NF-KB pathway. Oncol. Rep. 2013;29(2):571-78. [Crossref]  [PubMed] 
  19. Sethi G, Ahn KS, Aggarwal BB. Targeting nuclear factor-kappa B activation pathway by thymoquinone: role in suppression of antiapoptotic gene products and enhancement of apoptosis. Mol Cancer Res. 2008;6(6):1059-70. [Crossref]  [PubMed] 
  20. Ahmad A, Husain A, Mujeeb M, Khan SA, Najmi AK, Siddique NA, et al. A review on therapeutic potential of Nigella sativa: a miracle herb. Asian Pac J Trop Biomed. 2013;3(5):337-52. [Crossref]  [PubMed] 
  21. Galluzzi L, Senovilla L, Vitale I, Michels J, Martins I, Kepp O, et al. Molecular mechanisms of cisplatin resistance. Oncogene. 2012;31(15):1869-83. [Crossref]  [PubMed] 
  22. Shoieb AM, Elgayyar M, Dudrick PS, Bell JL,Tithof PK. In vitro inhibition of growth and induction of apoptosis in cancer cell lines by thymoquinone. Int J Oncol. 2003;22(1):107-13. [Crossref]  [PubMed] 
  23. Ng WK, Yazan LS, Ismail M. Thymoquinone from Nigella sativa was more potent than cisplatin in eliminating of SiHa ells via apoptosis with down-regulation of Bcl-2 protein. Toxicol in vitro. 2011;25:1392-98. [Crossref]  [PubMed] 
  24. Jafri SH, Glass J, Shi R, Zhang S, Prince M, Kleiner-Hancock H. Thymoquinone and cisplatin as a therapeutic combination in lung cancer: in vitro and in vivo. J Exp Clin Cancer Res. 2010;29:87. [Crossref]  [PubMed]  [PMC] 
  25. Nessa MU, Beale P, Chan C, Yu JQ, Huq F. Synergism from combinations of cisplatin and oxaliplatin with quercetin and thymoquinone in human ovarian tumour models. Anticancer Res. 2011;31(11): 3789-3797.
  26. Wilson AJ, Saskowski J, Barham W, Yull F, Khabele D. Thymoquinone enhances cisplatin-response through direct tumor effects in a syngeneic mouse model of ovarian cancer. J Ovarian Res. 2015;8:46. [Crossref]  [PubMed]  [PMC] 
  27. Alaufi OM, Noorwali A, Zahran F, Al-Abd AM, Al-Attas S. Cytotoxicity of thymoquinone alone or in combination with cisplatin (CDDP) against oral squamous cell carcinoma in vitro. Sci Rep. 2017;7(1):13131. [Crossref]  [PubMed]  [PMC] 
  28. Lei X, Lv X, Liu M, Yang Z, Ji M, Guo X, Dong W. Thymoquinone inhibits growth and augments 5-fluorouracil-induced apoptosis in gastric cancer cells both in vitro and in vivo. Biochem Biophys Res Commun. 2012;417(2):864-8. [Crossref]  [PubMed] 
  29. Effenberger-Neidnicht K, Schobert R. Combinatorial effects of thymoquinone on the anti-cancer activity of doxorubicin. Cancer Chemother Pharmacol. 2011;67(4):867-74. [Crossref]  [PubMed] 
  30. Ganji-Harsini S, Khazaei M, Rashidi Z, Ghanbari A. Thymoquinone could increase the efficacy of tamoxifen induced apoptosis in human breast cancer cells: an in vitro study. Cell J. 2016;18(2):245-54.

INDEX