Anti-proliferative effect of Thymoquinone on Raji cell line in Burkitt's lymphoma
Subject Areas : Journal of Animal BiologyMorteza Davoodi 1 , Shahriyar Saeediyan 2 , Reza Saghiri 3 , Zahra Zamani 4 , Gholamreza Bakhshi Khaniki 5
1 - Department of Biochemistry, Payame Noor university, Tehran, Iran
2 - Department of Biochemistry, Payame Noor university, Tehran, Iran
3 - Department of Biochemistry, Pasteur Institute of Iran, Tehran, Iran
4 - Department of Biochemistry, Pasteur Institute of Iran, Tehran, Iran
5 - Department of Plant Sciences, Payame Noor university, Tehran, Iran
Keywords: Nigella sativa, RT-PCR, Thymoquinone, Burkitt's lymphoma, Raji cells,
Abstract :
Medicinal plants are of interest due to their cheapness, accessibility and better acceptance by patients. One of these plants is black seed (Nigella sativa). In this study, the anti-proliferative effect of thymoquinone, which is the main component of black seed oil, is investigated on Raji cells. Raji cells are cancerous B lymphocytes that are seen in the germinal centers of Burkitt's lymphoma. In present study, Raji cells were treated with different dilutions of thymoquinone from 0 to 1000 μg/ml and the percentage of living cells was determined by trypan blue method and MTT test. Also, flow cytometry and Annexin V-FITC/PI kit were used to show the percentage of cells in different stages of growth. The expression of c-Myc gene, which is the most important altered gene in the development of Burkitt's lymphoma, was investigated by Real Time-PCR method. Statistical analysis was also done using SPSS 2020 software. This study showed that thymoquinone can inhibit the growth of Raji cells in a concentration-dependent and time-dependent manner. Thymoquinone, while suppressing the expression of c-Myc gene with a significant percentage, causes Raji cells to enter the stage of programmed death or apoptosis, and has the potential to be used as an adjunctive treatment for Burkitt's lymphoma.
1-Abukhader M.M. 2013. Thymoquinone in the clinical treatment of cancer: Fact or fiction? Pharmacognosy Reviews, 7(14): 117-120.
2- Adhikary S., Eilers M. 2005. Transcriptional regulation and transformation by Myc proteins. Molecular Cell Biology, 6(8):635- 645.
3- Banerjee S., Kaseb A.O., Wang Z., Kong D., Mohammad M., Padhye S., Sarkar F.H., Mohammad R.M. 2009. Antitumor activity of gemcitabine and oxaliplatin is augmented by thymoquinone in pancreatic cancer. Cancer Research, 69(13):5575-5583.
4- Beaulieu M.E., Castillo F., Soucek L. 2020. Structural and Biophysical Insights into the Function of the Intrinsically Disordered Myc Oncoprotein. Cells, 9(1038):1-27.
5- Burkitt D.P. 1983. The discovery of Burkitt’s lymphoma. Cancer, 51(10):1777-86.
6- Burkitt D.P. 1958. A sarcoma involving the jaws in African children. The British Journal of Surgery, 46(197):218-223.
7- Caponetti G., Bagg A. 2017. Demystifying the diagnosis and classification of lymphoma: a hematologist/oncologist’s guide to the hematopathologist’s galaxy. Journal of Community and Supportive Oncology, 15(1):43-48.
8- Chen H., Liu H., Qing G., 2018. Targeting oncogenic Myc as a strategy for cancer treatment. Signal Transduction and Targeted Therapy, 3(5):1-7.
9- Gilani A., Jabeen Q., Ullah Khan M. 2004. A review of medicinal uses and pharmacological activities of Nigella sativa. Pakistan Journal of Biological Sciences, 7: 441-451.
10- Hannan A., Rahman A., Sohag A.A., Uddin J., Dash R., Sikder M.H., Rahman S., Timalsina B., Munni Y.A., Sarker P.P., Alam M., Mohibbullah M., Haque N., Jahan I. Hossain T., Afrin T., Rahman M., Arif T.U., Mitra S., Oktaviani D.F., Khan K., Choi H.J., Moon I. Kim B. 2021. Black Cumin (Nigella sativa L.): A Comprehensive Review on Phytochemistry, Health Benefits, Molecular Pharmacology, and Safety. Nutrients, 13(6):1784.
11- Hu H.M., Kanda K., Zhang L., Boxer L.M. 2007. Activation of the c-Myc p1 promoter in Burkitt's lymphoma by the hs3 immunoglobulin heavy-chain gene enhancer. Leukemia, 21(4):747-753.
12- Kalkat M., Resetca D., Lourenco C., Chan P., Wei Y., 2018. MYC Protein Interactome profiling reveals functionally distinct regions that cooperate to drive tumorigenesis. Molecular Cell, 72(5):836-848.
13- Khan A., Tania M., Fu S., Fu J. 2017. Thymoquinone, as an anticancer molecule: from basic research to clinical investigation. Oncotarget, 8(31): 51907-51919.
14– Lewis D.W., Lilly S., Jones K.L. 2020. Lymphoma: Diagnosis and Treatment. Am Fam Physician, 101(1):34-41.
15- Ling Yong, CL., Wei Ow D.S., Tandiono T., Mei Heng L.L., Keung Chan K.K., Dieter Ohl C., Klaseboer E., Wan Ohl S., Hwa Choo A.B. 2014. Microbubble-mediated sonoporation for highly efficient transfection of recalcitrant human B- cell lines. Biotechnology Journal, 9(8):1081-1087.
16- Meerloo J.V., Kaspers G.J.L., Cloos J. 2015. Cell sensitivity assays: The MTT assay. Methods in Molecular Biology. Springer Ukraine, MIMB, (731): 237-244.
17- Misteli T. 2011. The inner life of the genome. HHS Author Manuscripts, 304(2): 66-73.
18- Mugnaini E.N., Ghosh N. 2016. Lymphoma. Prim Care. Lymphoma Research Gate, 43(4):661-675.
19- Nickavar B., Mojab F., Amoli M. 2003. Chemical composition of the fixed and volatile oils of Nigella sativa L. from Iran. National Library of Medicine, 58(9/10):629-631.
20- Padhye S., Banerjee S., Ahmad A., Mohammad R., Sarkar F.H. 2008. From here to eternity-the secret of Pharaohs: Therapeutic potential of black cumin seeds and beyond. Cancer Therapy, 6(b):495-510.
21- Potre O., Pescaru M., Sima A., Ionita L., Tudor R., Borsi E., Samfireag M., Potre C. 2021. Evaluation of the Relapse Risk and Survival Rate in Patients with Hodgkin Lymphoma: A Monocentric Experience. Medicina (Kaunas), 57(10): 1026.
22- Pourbakhsh H., Taghiabadi E., Abnous K., Hariri A.T., Hosseini S.M., Hosseinzadeh H. 2014. Effect of Nigella sativa fixed oil on ethanol toxicity in rats. Iran Journal Basic Medicine, 17(12):1020-31.
23- Pulvertaft, R.J.V. 1965. A study of malignant tumours in Nigeria by short-term tissue culture. Journal of Clinical Pathology, 18(3):261-273.
24- Rieger A.M., Nelson K.L., Konowalchuk J.D., Barreda, D.R. 2011. Modified annexin v/propidium iodide apoptosis assay for accurate assessment of cell death. Journal of Visualized Experiments, 24(50):2597.
25- Roix J.J., McQueen P.G., Munson P.J., Parada L.A., Misteli T. 2003. Spatial proximity of translocation-prone gene loci in human lymphomas. Nature Genetics, 34(3):287-291.
26- Schmitz R., Ceribelli M., Pittaluga S., Wright G., Staudt L.M., 2014. Oncogenic Mechanisms in Burkitt Lymphoma. Cold Spring Harbor Perspectives in Medicine, 4(2):a014282.
27- Singh S.K., Apata T., Gordetsky J.B., Singh R., 2019. Docetaxel Combined with Thymoquinone Induces Apoptosis in Prostate Cancer Cells via Inhibition of the PI3K/AKT Signaling Pathway. Cancers, 11(1390):1-13.
28- Wasylishen A., Penn L.Z. 2010. Myc The Beauty and the Beast. Genes Cancer, 1(6):532-541.
29- Yu Z., Wang R., Fok W.C., Cloes A., Salmon A.B., Perez V.I. 2015. Rapamycin and Dietary Restriction Induce Metabolically Distinctive Changes in Mouse Liver. Journals of Gerontology: Biological Sciences, 70(4):410-420.
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