Document Type : Research Paper

Authors

1 University of Baghdad - College of Science

2 Al-Mustansiriya University - Iraqi Center for Cancer and Medical Genetics Research

3 Al-Mustansiriya University - College of Science

Abstract

The present investigation represents a preliminary study of the effect of crude extracts (aqueous, ethanolic and hexanic) of date pits (Phoenix dactylifera L. cv. Zahdi) on two malignant cell lines (human laryngeal carcinoma-Hep2 and murine mammary adenocarcinoma-AMN3) and one normal cell line (rat embryo fibroblast-REF). The study also includes evaluation of the effect of these extracts on several cytogenetic parameters such as mitotic index (MI%), blast index (BI%) and chromosomal aberrations (CA) after in vitro culture of peripheral blood lymphocytes. This work also includes a study of the therapeutic potential of one of these extracts in the treatment of transplanted murine mammary adenocarcinoma in mice.The in vitro cell growth assay showed that there was time- and concentration-dependent cytotoxic effects of crude extracts of date pits on Hep2 and AMN3 cell lines. The highest significant effect of these extracts was achieved after 72 hrs of exposure with the highest concentration (10000 μg/ml). Ethanolic extract of pits (EP) caused growth inhibition percentage 89.4% , 93.4% for Hep2 and AMN3 respectively. However, 72 hrs exposure to EP at concentration of 10000 μg/ml caused slight inhibitory effect on REF cell line, reaching 17.7%.On the other hand, the crude extracts of pits caused significant reduction in the mitotic index and blast index of peripheral human lymphocytes, but without any structural or numerical chromosomal aberrations. Also these extracts neither replaced phytohemagglutinin (PHA) as mitogenic agent, nor colcemide as mitotic arresting agent at metaphase.The therapeutic doses of EP were determined according to LD50 in mice. The results indicated high effectiveness of this extract in a dose- and time-dependent manner. The highest therapeutic doses of EP (1 gm/kg B.wt.) showed the best therapeutic effect by reducing the tumor volume in mice to about 83.8%.The comparison of relative tumor volumes of different groups revealed highly significant differences among all treated groups and those of untreated (control) group.

Main Subjects

1- Greenlee, R.T. ; Hill-Hamon, M.B. ; Murray, T. and Thun, M. (2001). Cancer statistics 2001. CA. Cancer J. Clin., 51:15-36.
2- Jemal, A. ; Siegel, R. ; Ward, E. ; Murray, T. ; Xu, J. and Thun, M.J. (2007). Cancer Statistics, 2007. CA Cancer J. Clin., 57:43-66.
3- Ministry of Health. (2003). Results of Iraqi Cancer Registry 1999-2001, Iraqi Cancer Board, Baghdad, Iraq.  
4- Hellman, S. (1997). Principles of cancer management: Radiation therapy. In: V.T. DeVita ; S. Hellman and S.A. Rosenberg (Eds.), Cancer: principles and practice of oncology, (5th ed.), Lippincott-Raven Publishers, Philadelphia, pp.307-332.
5- Lopez-Lazaro, M. (2002). Flavonoids as anticancer agents: structure-activity relationship study. Curr. Med. Chem.-Anti-Cancer Agents, 2:691714.
6- DeVita, V.T. (1997). Principles of cancer management : chemotherapy. In:V.T. DeVita ; S. Hellman ; S.A. Rosenberg and L.H. Raven (ed.). Cancer principles and practice of oncology (5th ed.), Lippincott-Raven Publishers, Philadelphia, pp.333-348.
7- Kirn, D.H. (2000). Replication-selective microbiological agents : Fighting cancer with targeted germ warfare. J. Clinc. Invest., 105:837-839.
8- Katz, A.E. (2002). Flavonoid and botanical approaches to prostate health. The Journal of Alternative and Complementary Medicine, 8:813-821.
9- Azaizeh, H. ; Saad, B. ; Khalil, K. and Said, O. (2006). The state of the art of traditional arab herbal medicine in the eastern region of the mediterranean: A review. eCAM, 3:229-235.
10- Mukherjee, A.K. ; Basu, S. ; Sarkar, N. and Ghosha, A.C. (2001). Advances in cancer therapy with plant based natural products. Current Medicinal Chemistry, 8:1467-1486.
11- Jiménez-Medina, E. ; Garcia-Lora, A. ; Paco, L. ; Algarra, I. ; Collado, A. and Garrido, F. (2006). A new extract of the plant Calendula officinalis produces a dual in vitro effect: cytotoxic anti-tumor activity and lymphocyte activation. BMC Cancer, 6:119-132.
12- Yaseen, N.Y.; Hussein, S.M.; Saleh, F.S. and Mohammad, M.H. (2008). Plant and biological extracts in cancer therapy. Iraqi Center for Cancer and Medical Genetics Research(ICCMGR), Baghdad, Iraq.
13- Ahmed, I.A. ; Ahmed, A.W.K. and Robinson, R.K. (1995). Chemical composition of date varieties as influenced by the stage of ripening. Food Chemistry, 54:305-309.
14- Al-Qarawi, A.A. ; Ali, B.H. ; Al-Mougy, S.A. and Mousa, H.M. (2003). Gastrointestinal transit in mice treated with various extracts of date (Phoenix dactylifera L.). Food and Chemistry Technology, 41: 37-39.
15- Besbes, S. ; Blecker, C. ; Deroanne, C. ; Lognay, G. ; Drira, N.E. and Attia, H. (2004). Quality characteristics and oxidative stability of date seed oil during storage. Food Science and Technology International, 10: 333-338.
16- Dammak, I.; Abdallah, F.B.; Boudaya, S.; Besbes, S.; Keskes, L.; El Gaied, A.; Turki, H.; Attia, H. and Hentati, B. (2007). Date seed oil limit oxidative injuries induced by hydrogen peroxide in human skin organ culture. Biofactors, 29(2-3):137-45.
17- Ines, D.; Sonia, B.; Fatma, B.A.; Souhail, B.; Hamadi, A.; Hamida, T. and Basma, H. (2010). Date seed oil inhibits hydrogen peroxide-induced oxidative stress in human epidermal keratinocytes. Int. J. Dermatol., 49(3):262-8.
18- Kamel, B.S. ; Diab, M.F. ; Ilian, M.A. and Salman, A.J. (1981). Nutritional value of whole dates and date pits in broiler rations. Poultry Science, 60: 1005-1111.
19- Elgasim, E.A. ; Alyousef, Y.A. and Humeida A. M. (1995). Possible hormonal activity of date pits and flesh fed to meat animals. Food Chemistry, 52: 149-152.
20- Yousif, O.M. ; Osman, M.F. and Alhadrami, G.A. (1996). Evaluation of dates and date pits as dietary ingredients in Tilapia (Oreochromis aureus) diets differing in protein sources. Bioscience and Technology, 57: 81-85. 
21- Barreveld, W.H. (1993). Date palm products. FAO Agricultural Services Bulletin No. 101. Food and Agriculture Organization of the United Nations, Rome, Italy.
22- Hozumi, T. ; Yoshida, M. ; Ishida, Y. ; Mimoto, H. ; Sawa, J. ; Doi, K. and Kazumi, T.(1995). Long-term effects of dietary fiber supplementation on serum glucose and lipoprotein  levels in diabetic rats fed a high cholesterol diet. Endocr. J., 42:187-192.
23- Vuksan, V. ; Jenkins, D. ; Spadafora, P. ; Sievenpiper, J.L. ; Owen, R. ; Vidgen, E.; Brighenti, F. ; Josse, R. ; Leiter, L.A. and Bruce-Thompson, C.(1999). Konjac-mannan (glucamannan) improves glycemia and other associated risk factors for coronary heart diseases in type 2 diabetes. A randomized controlled metabolic trial. Diabetes Care, 22:913-919.
24- Walsh, D.E. ; Yaghoubian, V. and Behforooz, A. (1984). Effect of glucomannan on obese patients: a clinical study. Int.  J.  Obes., 8: 289-293.
25- Erskine, W.; Moustafa, A.T.; Osman, A.E.; Lashine, Z.; Nejatian, A.; Tamer Badawi, T. and Ragy, S.M.. Date Palm in the GCC countries of the Arabian Peninsula. Available at: {http://www.icarda.org/aprp/Datepalm/introduction/intro-body.htm}.
26- Harborne, J.B. ; Marbay, T.J. and Mabray, H. (1975). Physiology and function of flavonoids. Academic Press, New York, pp.970.
27- Simandi, B. ; Kery, A. ; Kristo, S.T. ; Andras, C. ; Prechi, A. and Fekete, J. (2001). Supercritical fluid extraction of non-volatile terpenoids from medicinal plants. Acta. Pharm. Hung.,71: 318-324.
28- Freshney, R.I. (2000). Culture of animal cells: A manual for basic technique (4th ed.). Wiley-liss, A John Wiley & Sons Inc. publication, New York.
29- Freshney, R.I. (1994). Culture of animal cells: A manual for basic technique (3th ed.). Wiley-liss, A John Wiley & Sons Inc. publication, New York.
30- Mahony, D.E. ; Gilliat, T. ; Dawson, S. ; Stockdale, E. and Lee, S.H. (1989). Vero cell assay for rapid detection of Clostridium perfringens enterotoxins. Applied and Environmental Microbiology. Pp:2141-2143.
31- Abdul-Majeed, M.R. (2000). Induction and characterization of SU.99 plasmacytoma cell line and its effect on mice immune response. Ph.D. Thesis, Nahrain University, Baghdad.
32- Betancur-Galvis, L.A. ; Saez, J. ; Granades, H. ; Salazer, A. and Ossa, J.E. (1999). Antitumor and antiviral activity of Colombian medicinal plant extracts. Mem. Inst. Oswaldo Cruz, 94: 531-535.
33- Yaseen, N.Y. ; Tawfiq, M.S. ; Hamadi, A.A. and Estivan, A.G. (1998). Cytogenetic studies on patient with chronic mylocytic leukemia. Med. J. Tikrit University, 4:5-9.
34- Yaseen, N.Y. (1990). Cytogenetic study on human colorectal cancer cell. Ph.D. Thesis, University of Sheffield.
35- Stites, D. (1979). Laboratory methods of detection cellular immune function. In: Fundberg, H.H. ; Stites, D. and Coldwell, J. (ed). Basic and clinical immunuology, LAGE, Medical Publication, California, pp. 318-322.
36- Shubber, E.K. and Al-Allak, B.M. (1986). Spontaneous chromosomal aberration and SCE in human lymphocytes : Effects of culture conditions. Nucleus, 22: 92-98.
37- ISCN. (1995). An international system for human cytogenetic nomenclature (1995). Mitelman, F.(ed.), S. Karger Publishers, Inc., Basel, Switzerland.
38- Dixon, W.J. (1980). Efficient analysis of experimental observations. Ann. Rev. Pharmacol. Toxicol., 20: 441-462.
39- Shibata, T. ; Giaccia, A.J. and Brown, M. (2002). Hypoxia-inducible regulation of a prodrug-activating enzyme for tumor-specific gene therapy. Neoplasia, 4:40-8.
40- Grote, D. ; Russell, S.J. ; Cornu, T.I. ; Cattaneo, R. ; Vile, R. ; Poland, G.A. and Fielding, A.K. (2001). Live attenuated measles virus induces regression of human lymphoma xenografts in immunodeficient mice. Blood, 97(12):3746-3754.
41- Blumenthal, R.M. ; Sharkey, R.M. ; Natale, A.M. ; Kashi, R. ; Wong, G. and Goldenberg, D.M. (1994). Comparison of equitoxic radio-immunotherapy and chemotherapy in treatment of human colonic cancer xenografts. Cancer Res., 54: 142-151.
42- Phuangsab, A. ; Lorence, R.M. ; Reichard, K.W. Peeples, M.E. and Walter, R.J. (2001). Newcastle disease virus therapy of human tumor xenografts : antitumor effects of local or systemic administration. Cancer Letters, 172: 27-36.
43 - Shubber, E.K. ; Amin, N.S. and El-Adhami, B.H. (1998). Cytogenetic effects of copper-containing intrauterine contraceptive device (IUCD) on blood lymphocytes. Mutat. Res., 417: 57-63.
44- Miller, O.J. and Therman, E. (2001). Human chromosome, (4th ed.), Springer-Verlag Inc., New York, pp.4.
45- Amany, M. M. B.; Shaker, M. A. and Abeer, A. K. (2012). Antioxidant activities of date pits in a model meat system. International Food Research Journal,19(1): 223-227
46- Neto, C.C. (2007). Cranberry and its phytochemicals: A review of in vitro anticancer studies. J. Nutr., 137: 186S-193S.
47- Bagchi, D. ; Sen, C.K. ; Bagchi, M. and Atalay, M. (2004). Anti-angiogenic, antioxidant and anticarcinogenic properties of a novel anthocyanin-rich berry extract formula. Biochemistry (Mosc), 69: 75-80.
48- Atalay, M. ; Gordillo, G. ; Roy, S. ; Rovin, B. ; Bagchi, D. ; Bagchi, M. and Sen, C.K. (2003). Anti-angiogenic property of edible berry in a model of hemangioma. FEBS Lett., 544: 252-257.