Asian Pacific Journal of Cancer Prevention

Asian Pacific Journal of Cancer Prevention (아시아태평양암예방학회)
권호 표지
DOI QR코드

DOI QR Code

Roles of p53 and Caspases in Induction of Apoptosis in MCF-7 Breast Cancer Cells Treated with a Methanolic Extract of Nigella Sativa Seeds

  • Alhazmi, Mohammed I. (Molecular Cancer Biology Research Lab., Dept. of Food Science and Nutrition, College of Food and Agricultural Sciences, King Saud University) ;
  • Hasan, Tarique N. (Molecular Cancer Biology Research Lab., Dept. of Food Science and Nutrition, College of Food and Agricultural Sciences, King Saud University) ;
  • Shafi, Gowhar (Molecular Cancer Biology Research Lab., Dept. of Food Science and Nutrition, College of Food and Agricultural Sciences, King Saud University) ;
  • Al-Assaf, Abdullah H. (Molecular Cancer Biology Research Lab., Dept. of Food Science and Nutrition, College of Food and Agricultural Sciences, King Saud University) ;
  • Alfawaz, Mohammed A. (Molecular Cancer Biology Research Lab., Dept. of Food Science and Nutrition, College of Food and Agricultural Sciences, King Saud University) ;
  • Alshatwi, Ali A. (Molecular Cancer Biology Research Lab., Dept. of Food Science and Nutrition, College of Food and Agricultural Sciences, King Saud University)
  • Published : 2014.12.18
Download PDF
⟨ Previous Next ⟩

Abstract

Background: Nigella Sativa (NS) is an herb from the Ranunculaceae family that exhibits numerous medicinal properties and has been used as important constituent of many complementary and alternative medicines (CAMs). The ability of NS to kill cancer cells such as PC3, HeLa and hepatoma cells is well established. However, our understanding of the mode of death caused by NS remains nebulous. The objective of this study was to gain further insight into the mode and mechanism of death caused by NS in breast cancer MCF-7 cells. Materials and Methods: Human breast cancer cells (MCF-7) were treated with a methanolic extract of NS, and a dose- and time-dependent study was performed. The $IC_{50}$ was calculated using a Cell Titer $Blue^{(R)}$ viability assay assay, and evidence for DNA fragmentation was obtained by fluorescence microscopy TUNEL assay. Gene expression was also profiled for a number of apoptosis-related genes (Caspase-3, -8, -9 and p53 genes) through qPCR. Results: The $IC_{50}$ of MCF-7 cells was $62.8{\mu}L/mL$. When MCF-7 cells were exposed to $50{\mu}L/mL$ and $100{\mu}L/mL$ NS for 24h, 48h and 72h, microscopic examination (TUNEL assay) revealed a dose- and time-dependent increase in apoptosis. Similarly, the expression of the Caspase-3, -8, -9 and p53 genes increased significantly according to the dose and time. Conclusions: NS induced apoptosis in MCF-7 cells through both the p53 and caspase pathways. NS could potentially represent an alternative source of medicine for breast cancer therapy.

Keywords

References

  1. Al-Hader A, Aqel M, Hasan Z (1993). Hypoglycemic effects of the volatile oil of Nigella sativa seeds. Int J Pharmacogn, 31, 96-100. https://doi.org/10.3109/13880209309082925
  2. Alshatwi AA, Shafi G, Hasan TN, Syed NA, Khoja KK (2013). Fenugreek induced apoptosis in breast cancer MCF-7 cells mediated independently by fas receptor change. Asian Pac J Cancer Prev, 14, 5783-8. https://doi.org/10.7314/APJCP.2013.14.10.5783
  3. Chopra RN, Nayar SL, Chopra IC (1956). Glossary of Indian Medicinal Plants. CSIR, New Delhi, India, p. 175.
  4. Czabotar PE, Lessene G, Strasser A, Adams JM (2014). Control of apoptosis by the BCL-2 protein family: implications for physiology and therapy. Nat Rev Mol Cell Biol, 15, 49-63.
  5. Earnshaw WC, Martins LM, Kaufmann SH (1999). Mammalian caspases Structure, activation, substrates, and functions during apoptosis. Annu Rev Biochem, 68, 383-424. https://doi.org/10.1146/annurev.biochem.68.1.383
  6. El Sharkawi FZ, El Shemy HA, Khaled HM (2014). Possible anticancer activity of rosuvastatine, doxazosin, repaglinide and oxcarbazepin. Asian Pac J Cancer Prev, 15, 199-203. https://doi.org/10.7314/APJCP.2014.15.1.199
  7. El-Daly ES (1998). Protective effect of cysteine and Vitamin E, Crocus sativus and Nigella sativa extracts on cisplatininduced toxicity in rats. J Pharm Belg, 53, 87-95.
  8. El-Kamali HH, Ahmed AH, Mohammed AAM (1998). Antibacterial properties of essential oils from Nigella sativa seeds, Cymbopogon citratus leaves and Pulicaria undulata aerial parts. Fitoterapia, 69, 77-8.
  9. El-Shabrawy OA, Nada SA (1996). Biological evaluation of multicomponent tea used as hypoglycemic in rats. Fitoterapia, 67, 99-102.
  10. El-Tahir KEH, Ashour MMS, Al-Harbi MM (1993). The respiratory effects of the volatile oil of the black seed (Nigella sativa) in guinea-pigs: elucidation of the mechanism(s) of action. Gen Pharmacol, 24, 1115-22. https://doi.org/10.1016/0306-3623(93)90358-5
  11. Florea AM, Busselberg D (2011). Cisplatin as an anti-tumor drug: cellular mechanisms of activity, drug resistance and induced side effects. Cancer, 3, 1351-71. https://doi.org/10.3390/cancers3011351
  12. Fridman JS, Lowe SW (2003). Control of apoptosis by p53. Oncogene, 22, 9030-40. https://doi.org/10.1038/sj.onc.1207116
  13. Gali-Muhtasib H, Diab-Assaf M, Boltze C, et al (2004). Thymoquinone extracted from black seed triggers apoptotic cell death in human colorectal cancer cells via a p53dependent mechanism. Int J Oncol, 25, 857-66.
  14. Goreja WG (2003). Black Seed: Nature's Miracle Remedy. New York, NY 7 Amazing Herbs Press.
  15. Hasan TN, Ahmed SN, Aalam SMM, Kumar C and Shafi G (2007). Evaluation of cichorium extract for the growth supporting property in rat hepatocyte primary culture. Asian J Plant Sci, 6, 431-4. https://doi.org/10.3923/ajps.2007.431.434
  16. Hasan TN, Grace BL, Shafi G, Al-Hazzani AA, Alshatwi AA (2011). Anti-proliferative effects of organic extracts from root bark of Juglans Regia L. (RBJR) on MDA-MB-231 human breast cancer cells: role of Bcl-2/Bax, caspases and Tp53. Asian Pac J Cancer Prev, 12, 525-30.
  17. Hasan TN, Shafi G, Syed NA, et al (2013). Methanolic extract of Nigella sativa seed inhibits SiHa human cervical cancer cell proliferation through apoptosis. Nat Prod Commun, 8, 213-6.
  18. Hengartner MO (2000). The biochemistry of apoptosis. Nature, 407, 770-6. https://doi.org/10.1038/35037710
  19. Houghton PJ, Zarka R, De las Heras B, Hoult JRS (1995). Fixed oil of Nigella sativa and derived thymoquinone inhibit eicosanoid generation in leukocytes and membrane lipid peroxidation. Planta Med, 61, 33-6. https://doi.org/10.1055/s-2006-957994
  20. Ivankovic S, Stojkovic R, Jukic M, Milos M, Jurin M (2006). The anti tumor activity of thymoquinone and thymohydoquinone in vitro and invivo. Exp Oncol, 28, 220-4.
  21. Khanna T, Zaidi FA, Dandiya PC (1993). CNS and analgesic studies on Nigella sativa. Fitoterapia, 64, 407-10.
  22. Kma L (2013). Roles of plant extracts and constituents in cervical cancer therapy. Asian Pac J Cancer Prev, 14, 3429-36 https://doi.org/10.7314/APJCP.2013.14.6.3429
  23. Kundu J, Kim DH, Kundu JK, Chun KS (2014). Thymoquinone induces heme oxygenase-1 expression in HaCaT cells via Nrf2/ARE activation: Akt and $AMPK{\alpha}$ as upstream targets. Food Chem Toxicol, 65, 18-26. https://doi.org/10.1016/j.fct.2013.12.015
  24. Mahmoud MR, El-Abhar HS, Saleh S (2002). The effect of Nigella sativa oil against the liver damage induced by Schistosoma mansoni infection in mice. J Ethnopharmacl, 79, 1-11. https://doi.org/10.1016/S0378-8741(01)00310-5
  25. Mangal M, Sagar P, Singh H, Raghava GP, Agarwal SM (2013). NPACT: Naturally Occurring Plant-based Anti-cancer Compound-Activity-Target database. Nucleic Acids Res, 41, 1124-9. https://doi.org/10.1093/nar/gks1045
  26. Motomura M, Kwon KM, Suh SJ, et al (2008). Propolis induces cell cycle arrest and apoptosis in human leukemic U937 cells through Bcl-2/Bax regulation. Environ Toxicol Pharmacol, 26, 61-7. https://doi.org/10.1016/j.etap.2008.01.008
  27. Mouhajir F, Pedersen JA, Rejdali M, Towers GHN (1999). Antimicrobial thymohydroquinones of Moroccan Nigella sativa seeds detected by electron spin resonance. Pharm Biol, 37, 391-5. https://doi.org/10.1076/phbi.37.5.391.6052
  28. Mutabagani A, El-Mahdy SAM (1997). A study of the antiinflammatory activity of Nigella sativa L. and thymoquinone in rats. Saudi Pharmaceutical J, 5, 110-3.
  29. Nadakarni KM (1976). Corcus sativus, Nigella sativa. In: Nadkarni, K.M. (Ed.), Indian materia medica. popular Prakashan, Bombay, India; p 386-411.
  30. Nair SC, Salomi MJ, Panikkar B, Panikkar KR (1991). Modulatory effects of Crocus sativus and Nigella sativa extracts on cisplatin-induced toxicity in mice. J Ethnopharmacol, 31, 75-83. https://doi.org/10.1016/0378-8741(91)90146-5
  31. Pekar O, Molotski N, Savion S, et al (2007). p53 regulates cyclophosphamide teratogenesis by controlling caspases 3, 8, 9 activation and NF-kappaB DNA binding. Reproduction, 134, 379-88. https://doi.org/10.1530/REP-07-0086
  32. Pommier Y, Sordet O, Antony S, Haywrd RL, Kohn KW (2004). Apoptosis defects and chemotherapy resistance: molecular interaction maps and networks. Oncogene, 23, 2934-49. https://doi.org/10.1038/sj.onc.1207515
  33. Salim LZ, Mohan S, Othman R, Abdelwahab SI, Kamalidehghan B, Sheikh BY, et al. (2013). Thymoquinone induces mitochondria-mediated apoptosis in acute lymphoblastic leukaemia in vitro. Molecules, 18, 11219-40. https://doi.org/10.3390/molecules180911219
  34. Salomi NJ, Nair SC, Jayawardhanan KK (1992). Antitumour principles from Nigella sativa seeds. Cancer Lett, 63, 41-6. https://doi.org/10.1016/0304-3835(92)90087-C
  35. Salomi NJ, Nair SC, Jayawardhanan, KK, Vargghese CD, Pantikkar KR (1992). Anti-tumor principles from Nigella sativa seeds. Cancer Lett, 63, 41-6. https://doi.org/10.1016/0304-3835(92)90087-C
  36. Salomi NJ, Nair SC, Panikkar KR (1991). Inhibitory effects of Nigella sativa and Saffron (Crocus sativus) on chemical carcinogenesis in mice. Nutr Cancer, 16, 67-72. https://doi.org/10.1080/01635589109514142
  37. Shafi G, Hasan TN, Sayed NA (2008). Methanolic extract of Nigella sativa seeds is potent clonogenic inhibitor of pc3 cells. Int J Pharmacol, 4, 477-81. https://doi.org/10.3923/ijp.2008.477.481
  38. Shafi G, Munshi A, Hasan TN, et al (2009). Induction of apoptosis in HeLa cells by chloroform fraction of seed extracts of Nigella sativa. Cancer Cell Int, 9, 29. https://doi.org/10.1186/1475-2867-9-29
  39. Sonis ST (2011). Oral mucositis. Anticancer Drugs, 22, 607-12. https://doi.org/10.1097/CAD.0b013e3283462086
  40. Takaoka A, Hayakawa S, Yanai H, et al (2003). Integration of interferon-${\alpha}/{\beta}$ signalling to p53 responses in tumour suppression and antiviral defense. Nature, 424, 516-23. https://doi.org/10.1038/nature01850
  41. Thabrew MI, Mitry RR, Morsy MA, Hughes RD (2005). Cytotoxic effects of a decoction of Nigella sativa, Hemidesmus indicus and Smilax glabra on human hepatoma HepG2 cells. Life Sci, 77, 1319-30. https://doi.org/10.1016/j.lfs.2005.01.022
  42. Yesilada E, Honda G, Sezik E, et al (1995). Traditional medicine in Turkey. V. Folk medicine in the inner Taurus Mountains. J Ethnopharmacol, 46, 133-52. https://doi.org/10.1016/0378-8741(95)01241-5
  43. Yuan JS Reed A, Chen F, Stewart Jr CN (2006). Statistical analysis of real-time PCR data, BMC Bioinformatics, 7, 85. https://doi.org/10.1186/1471-2105-7-85
  44. Zaoui A, Cherrah Y, Alaoui K, et al (2002). Effects of Nigella sativa fixed oil on blood homeostasis in rat. J Ethnopharmacol, 79, 23-6. https://doi.org/10.1016/S0378-8741(01)00342-7
  45. Zhang H, Lei Y, Yuan P, et al (2014). ROS-mediated autophagy induced by dysregulation of lipid metabolism plays a protective role in colorectal cancer cells treated with gambogic acid. PLoS One, 9, 96418. https://doi.org/10.1371/journal.pone.0096418

Cited by

  1. Silibilin-Induces Apoptosis in Breast Cancer Cells by Modulating p53, p21, Bak and Bcl-xl Pathways vol.16, pp.5, 2015, https://doi.org/10.7314/APJCP.2015.16.5.2087
  2. Dietary Natural Products for Prevention and Treatment of Breast Cancer vol.9, pp.7, 2017, https://doi.org/10.3390/nu9070728
  3. Changes in apoptosis-related gene expression profiles in cancer cell lines exposed to usnic acid lichen secondary metabolite vol.41, pp.13036092, 2017, https://doi.org/10.3906/biy-1609-40
  4. Effect of CCNB1 silencing on cell cycle, senescence, and apoptosis through the p53 signaling pathway in pancreatic cancer vol.234, pp.1, 2019, https://doi.org/10.1002/jcp.26816