اثر باز دارنده رشد عصاره شبدر قرمز بر سلولهای سرطانی راجی در لنفوم بورکیت
Subject Areas : Journal of Medicinal Herbs, "J. Med Herb" (Formerly known as Journal of Herbal Drugs or J. Herb Drug)مرتضی داودی 1 , شهریار سعیدیان 2 , رضا صغیری 3 , زهرا زمانی 4 , غلام رضا بخشی خانیکی 5
1 - گروه بیوشیمی، دانشگاه پیام نور، تهران، ایران;
2 - گروه بیوشیمی، دانشگاه پیام نور، تهران، ایران;
3 - انستیتو پاستور ایران، تهران، ایران;
4 - انستیتو پاستور ایران، تهران، ایران;
5 - گروه زیست شناسی، دانشگاه پیام نور، تهران، ایران;
Keywords: ID3, Red Clover, p53, Burkitt's lymphoma, c-MYC, Raji cells,
Abstract :
Background & Aim: In folk medicine, red clover (Trifolium pratense) use to treat some diseases such as whooping cough, asthma, eczema, and some ophthalmic dysfunction. Due to its phytoestrogens and antioxidant-rich compounds, it is recommended to alleviate some cancers. This paper was conducted to study the anti-cancer and apoptosis effects of red clover extract (RCE) on the Raji cell line in Burkitt's lymphoma.Experimental: Red clover lyophilized powder (CAS = 85085-25-2) with a fixed ratio of Formononetin, Biochanin A, Daidzein, and Genistein was acquired from Sigma Aldrich German company. Raji cells were obtained from the Pasteur Institute of Iran-Tehran. Raji cells were treated with different concentrations of RCE, and viable cells were measured at an interval of 24, 48, and 72 hours incubation in three days by MTT assay. Agarose gel electrophoresis was used to determine the DNA degradation. Annexin V-FITC/PI kit and flow cytometry assay were used to determine the percent of apoptotic and necrotic treated Raji cells. Also, expression changes in cMYC, ID3, and P53 genes were measured by real-time PCR. Results: Red clover extract can prevent in vitro proliferation of the Raji cells in a time and dose-dependent manner. RCE, as well as c-MYC gene suppression and induction of ID3 and P53 genes, enters the Raji cells in the apoptosis stages with an acceptable percentage and has complication cure potential in Burkitt's lymphoma. Recommended applications/industries: The extract used in this study combines four components. The individual evaluation effects of Formononetin, Biochanin A, Daidzein, and Genistein on Raji cells are recommended.
Akbaribazm, M. Khazaei, M.R. and Khazaei, M. 2020. Trifolium pratense L. (red clover) extract and Doxorubicin synergistically inhibit proliferation of 4T1 breast cancer in tumor-bearing BALB/c mice through modulation of apoptosis and increase antioxidant and anti-inflammatory related pathways. Food Science and Nature, 8(8): 4276-4290.
Beaulieu, M.E., Castillo, F. and Soucek, L. 2020. Structural and biophysical insights into the function of the intrinsically disordered MycOncoprotein. Cells, 9(1038): 1-27.
Caponetti, G. and Bagg, A. 2017. Demystifying the diagnosis and classification of lymphoma: hematologist/oncologist's guide to the hematopathologist's galaxy. Journal of Community and Supportive Oncology, 15(1): 43-48.
Chen, D.R. and Cohen, P.L. 2012. Living life without B cells: is repeated B-cell depletion a safe and effective long-term treatment plan for rheumatoid arthritis?. National Library of Medicine,7(2): 159-166.
Dorvignit, D., Palacios, J.L., Merino, M., Hernandez, T., Sosa, K., Casaco, A., Lopez-Requena, A. and Mateo de Acosta, C. 2012. Expression and biological characterization of an anti-CD20 biosimilar candidate antibody. National Library of Medicine Mabs, 4(4): 488-496.
Green, M. R. and Sambrook, J. 2019. Analysis of DNA by agarose gel electrophoresis. Cold Spring Harbor Protocols, 2019(1): 1-3.
Hu, H-M., Kanda, K., Zhang, L. and Boxer, L.M. 2007. Activating the c-Mycp1 promoter in Burkitt's lymphoma by the hs3 immunoglobulin heavy-chain gene enhancer. Leukemia, 21(4): 747-53.
Janas, E., Priest, R., Wilde, J.I. White, J.H. and Malhotra, R. 2005. Rituxan (anti-CD20 antibody)-induced translocation of CD20 into lipid rafts is crucial for calcium influx and apoptosis. National Library of Medicine, 139(3): 436-446.
Kianinodeh, F., Tabatabaei, S.M., Alibakhshi, A., Gohari, M. and Tari, K. 2017. Anti-tumor effects of essential oils of red clover and ragweed on Mcf-7 breast cancer cell line. Scientific Information Database, 1(4): 17-23.
Kucuhboyaci, N., Kadioglu, O., Adiguzel, N., Tamer, U., Guvenc, A. and Bani, B. 2013. Determination of isoflavone content by HPLC-UV method and in vitro antioxidant activity of red clover (Trifolium pratense L). Turk Journal of Pharmacology Science, 10(3): 463-472.
Lewis, D.W., Lilly, S. and Jones, K.L. 2020. Lymphoma: diagnosis and treatment. National Library of Medicine, Am Fam Physician. 101(1): 34-41.
Majtnerova, P. and Rousar, T. 2018. An overview of apoptosis assays detecting DNA fragmentation. National Library of Medicine, 45(5): 1469-1478.
Mallick, I. 2022. B-cells in your immune system. Verywell Health, Updated on march 23, 2022. medically reviewed by Douglas A. and Nelson M.D. Available on internet at: https://www,verywellhealth.com> how-dose- imm.
Mannella, P. 2012. Effects of red clover extract on breast cancer cell migration and invasion. Gynecology and Endocrinology, 28(1):29-33.
Meerloo, J. V., Kaspers, G.J.L. and Cloos, J. 2015. Cell sensitivity assays: The MTT assay. Methods in molecular biology. Springer Ukraine, MIMB, (731): 237-244.
Misteli, T. 2011. The inner life of the genome. National Library of Medicine, HHS Author Manuscripts, 304(2): 66-73.
Mugnaini, E.N. and Ghosh, N. 2016. Lymphoma. Prim Care. Lymphoma Research Gate, 43(4): 661-675.
Nelsen, J., Ulbricht, C., Paul Barrette, E., Sollars, D., Tsourounis, C., Rogers A., Basch, S., Hashemi, S., Bent, S. and Basch, E. 2002. Red clover (Trifolium pretense) monograph: A clinical decision support tool. Journal of Herbal Pharmacotherapy, 2(3): 49-72.
Omoto, Y. and Iwase, H. 2015. Clinical significance of estrogen receptor β in breast and prostate cancer from biological aspects. Cancer Sciences, 106(4): 337-343.
Pierpont, T.M., Limper, C.B. and Richards, K.L. 2018. Past, present, and future of Rituximab—The world's first oncology monoclonal antibody therapy. Available on internet at Frontiers in Oncology, https://doi.org/10.3389/fonc.2018.00163.
Potre, O., Pescaru, M., Sima, A., Ionita, L., Tudor, R., Borsi, E., Samfireag, M. and Potre, C. 2021. Evaluation of the relapse risk and survival rate in patients with hhodgkin lymphoma: A monocentric experience. National Library of Medicine, Medicina (Kaunas), 57(10): 1026.
Pulvertaft, R.J. 1965. A study of malignant tumours in Nigeria by short-term tissue culture. National Library of Medicine, 18(3): 261-273.
Rieger, A.M. Nelson, K.L., Konowalchuk, J.D. and Barreda, D.R. 2011. Modified Annexin V/Propidium Iodide Apoptosis Assay For Accurate Assessment of Cell Death. Journal of Visualized Experiments, 24(50): 2597.
Rohde, M., Bonn, B.R., Zimmermann, M., Lange, J., Moricka, A., Klapper, W., Oschlies, I., Szczepanowski, M., Nagel, I. and Schrappe, M. 2017. Relevance of ID3-TCF3-CCND3 pathway mutations in pediatric aggressive B-cell lymphoma treated according to the non-Hodgkin lymphoma. National Library of Medicine, Haematologica, 102(6): 1091-1098.
Roix, J.J., McQueen, P.G., Munson, P.J., Parada, L.A. and Misteli, T. 2003. Spatial proximity of translocation-prone gene loci in human lymphomas. Nature Genetics, 34(3): 287-91.
Smardove, J., Grochova, D., Moulis, M. and Fabian, P. 2008. An unusual p53 mutation was detected in Burkitt's lymphoma: 30 bp duplication. Oncology Reports, 20(4):773-778.
Wasylishen, A. and Penn, L. Z. 2010. Myc: the beauty and the beast. Genes Cancer, 1(6): 532-41.
Yakimchuk, K., Sharif Hasni, M., Guan, J., Chao, M.P., Sander, B. and Okret, S. 2014. Inhibition of lymphoma vascularization and dissemination by estrogen receptor β agonists. Blood, 123(13): 2054-2061.
Yuan, J.D., Reed, A., Chen, F. and Stewart, N. 2006. Statistical analysis of real-time PCR data. MMC Bioinformatics. 7, 85(2006).
Zaklos-Szyda, M. and Budryn, G. 2020. The effects of Trifolium pratense L. sprouts' phenolic compounds on cell growth and migration of MDA-MB-231, MCF-7, and HUVEC cells. Nutrients, 12(1): 1-26.