POTENTIAL ANTIOXIDANT AND CYTOTOXIC ACTIVITIES OF ARECA NUT (ARECA CATECHU LINN.) EXTRACT IN HUMAN ORAL SQUAMOUS CELL CARCINOMA AND KERATINOCYTE CELLS
Â Objectives: Areca (betel, Areca catechu) nuts have been reported to show high contents of phenolic and flavonoid compounds with associated antioxidant activity. This study was undertaken to assess the anticancer potential of areca nut extract on human squamous carcinoma (HSC)-3, HSC-2, and human keratinocyte (HaCat) cancer cell lines, and to elucidate the mechanism of its action.
Methods: In this study, the areca nut extract was prepared in 96% ethanol and analyzed for the total phenolic and flavonoid content, 2,2-diphenyl-1- picrylhydrazyl (DPPH) assay, and 5-(3-carboxymethoxyphenyl)-2-(4,5-dimenthylthiazoly)-3-(4-sulfophenyl)tetrazolium inner salt assay.
Results: The indicated total phenolic content of the areca nut extract was 80.3 mg tannic acid equivalent/g and total flavonoid content 238.5 mg catechin equivalent/g. The extract showed good scavenging activity against DPPH radical (efficient concentration 15.95Â±0.84 Î¼g/ml). The extract showed higher toxicity on HSC-3 than HSC-2 cells, IC50 was 164.06 Î¼g/ml in HSC-3 versus 629.50 Î¼g/ml in HSC-2. The extract had no cytotoxic effect on HaCat cells.
Conclusions: The antioxidant activity of the areca nut extract might be associated with the presence of high content of phenolic and flavonoid compounds. The extract showed selective cytotoxicity against HSC-3 and HSC-2 cells but not on HaCat cells. Instead, the extract induced high rate of proliferation in HaCat cells.
2. Ascani G, Balercia P, Messi M, Lupi L, Goteri G, Filosa A, et al. Angiogenesis in oral squamous cell carcinoma. Acta Otorhinolaryngol Ital 2005;25(1):13-7.
3. Jemal A, Siegel R, Ward E, Hao Y, Xu J, Murray T, et al. Cancer statistics, 2008. CA Cancer J Clin 2008;58(2):71-96.
4. Siegel RL, Miller KD, Jemal A. Cancer statistics, 2015. CA Cancer J Clin 2015;65(1):5-29.
5. Neville BW, Damm DD, Allen CM, dan Bouquot JE. Squamous Cell Carcinoma in Oral and Maxillofacial Pathology. 2nd ed. Philadelphia, PA, Pennsylvania: Saunders; 2002. 6. Sreejamole KL, Radhakrishnan CK. Antioxidant and cytotoxic activities of ethyl acetat extract of the Indian green Perna viridis. Asian J Pharm Clin Res 2013;6(3):197-201.
7. Chitra T, Jayashree S, Rathinamala J. Evaluation of anticancer of Vetiveria zizanioides against breast cancer cell line. Int J Pharm Pharm Sci 2014;6(1):164-6.
8. Hamsar MN, Ismail S, Mordi M, Ramanathan S, Mansor SM. Antioxidant capacity and the effect of different parts of Areca catechu extracts on gluthatione-S-transferase activity in vitro. Free Radic Antioxid 2011;1(1):28-33.
9. Jaiswal P, Kumar P, Singh VK, Singh DK. Areca catechu L.: A valuable herbal medicine againts different health problems. Res J Med Plant 2011;5(2):145-52.
10. Kim BJ, Kim JH, Kim HP, Heo MY. Biological screening of 100 plant extracts for cosmetic use (II): Anti-oxidative activity and free radical scavenging activity. Int J Cosmet Sci 1997;19:299-307.
11. Lee KK, Choi JD. The effects of Areca catechu L extract on anti-inflammation and antimelanogenesis. Int J Cosmet Sci 1999;21(4):275-80.
12. Lee KK, Choi JD. The effects of Areca catechu L extract on anti-aging. Int J Cosmet Sci 1999;21(4):285-95.
13. Bhandare AM, Kshirsagar AD, Vyawahare NS, Hadambar AA, Thorve VS. Potential analgesic, anti-inflammatory and antioxidant activities of hydroalcoholic extract of Areca catechu L. nut. Food Chem Toxicol 2010;48(12):3412-7.
14. Velioglu YS, Mazza G, Gao L, Oomah BD. Antioxidant activity and total phenolics in selected fruits, vegetables, and grain products. J Agric Food Chem 1998;46(9):4113-7.
15. Ahsan H, Ali A, Ali R. Oxygen free radicals and systemic autoimmunity. Clin Exp Immunol 2003;131(3):398-404.
16. Zhang WM, Wei J, Chen WX, Zhang HD. The chemical composition and phenolic antioxidants of Areca (Areca catechu L.) seeds. Int Conf Agric Biosyst Eng 2011;1-2:16-22.
17. Huang PL, Chi CW, Liu TY. Effects of Areca catechu L. containing procyanidins on cyclooxygenase-2 expression in vitro and in vivo. Food Chem Toxicol 2010;48(1):306-13.
18. Xing Z, Jiao W, Zhuang H, Li MW, Fu DH. Antioxidant and cytotoxic phenolic compounds of Areca nut (Areca catechu L.). Chem Res Chin Univ 2010;26(1):161-4.
19. Roy S, Choudhury MD, Paul SB. Antioxidant potential of rhiome of Alocasia decipiens schott. Asian J Pharm Clin Res 2013;6(2):120-2.
20. Ramamoorthy P, Bono A. Antioxidant activity, total phenolic, and flavonoid content of Morinda citrifolia fruit extracts from various extraction processes. Int J Environ Sci Technol 2007;2(1):70-80.
21. Zhang WM, Huang WY, Chen WX, Han L, Zhang HD. Optimalization of extraction conditions of Areca seed polyphenols and evaluation of their antioxidant activities. Molecules 2014;19:16416-27.
22. Kamiya Y, Ohshima T. The individual cell properties of oral squamous carcinoma and tumor suppressor gene mutation. Oral Sci Int 2005;2(2):104-17.
23. Okamura M, Shimada J, Sakagami H. Comparative analysis of cell death induction by cisplatin and 5-FU in human oral squamous and hepatocellular carcinoma cell lines. Anticancer Res 2008;28:253-60.
24. Boukamp P, Petrussevska RT, Breitkreutz D, Hornung J, Markham A, Fusenig NE. Normal keratinization in a spontaneously immortalized aneuploid human keratinocyte cell line. J Cell Biol 1988;106(3):761-71.
25. Dewoto HR. The developing of Indonesia traditional herbal medicines in to phytopharmaca. Maj Kedokt Indon 2007;57(7):205-11.
26. Bhisey RA, Boucher BJ, Hsi-Chen TH, Gajalakshmi V, Gupta PC, et al. Monographs on the evaluation of carcinogenic risks to humans. Betel-Quid and Areca-Nut Chewing and Some areca-Nut Related Nitrosamines. Vol. 85. California: IARCS Monographs; 2004. p. 1-293.
27. Nagao T, Ikeda N, Warnakulasuriya S, Fukano H, Yuasa H, Yano M, et al. Serum antioxidant micronutrients and the risk of oral leukoplakia among Japanese. Oral Oncol 2000;36(5):466-70.
28. Carossa S, Pera P, Doglio P, Lombardo S, Colagrande P, Brussino L, et al. Oral nitric oxide during plaque deposition. Eur J Clin Invest 2001;31(10):876-9.
29. Nair U, Bartsch H, Nair J. Alert for an epidemic of oral cancer due to use of the betel quid substitutes gutkha and pan masala: A review of agents and causative mechanisms. Mutagenesis 2004;19(4):251-62.
30. Johnson NW, Jayasekara P, dan Amarasinghe HK. Squamous cell carcinoma and precursor lesions of the oral cavity: Epidemiology and aetiology. In: Periodontology. 2000. Vol. 57. Singapura: John Wiley and Sons; 2011.
31. Anttila A, Bhat RV, Bhond JA, Bhorgoff SJ, Bosch FX, Carlson GP, et al. Monographs on the evaluation of carcinogenic risks to humans. Some Traditional Herbal Medicines, Some Mycotoxins, Naphthalene and Styrene. Vol. 82. California: IARCS Monographs; 2002. p. 171-4.
32. Hannan A, Karan S, Chatterjee TP. A comparative study of in vitro antioxidant capacity of different extract of Areca seed collected from Areca catechu L plant grown in Assam. Int J Pharm Pharm 2012;4(2):420-7.
33. Tharakan B, Dhanasekaran M, Manyam BV. Antioxidant and DNA protecting properties of anti-fatigue herb Trichopus zeylanicus. Phytother Res 2005;19(8):669-73.
34. Kong Y, Ma W, Liu X, Zu Y, Fu Y, Wu N, et al. Cytotoxic activity of curcumin towards CCRF-CEM leukemia cells and its effect on DNA damage. Molecules 2009;14(12):5328-38.
35. Gibellini L, Pinti M, Nasi M, De Biasi S, Roat E, Bertoncelli L, et al. Interfering with ROS metabolism in cancer cells: The potential role of quercetin. Cancers (Basel) 2010;2(2):1288-311.
36. Sakano K, Kawanishi S. Metal-mediated DNA damage induced by curcumin in the presence of human cytochrome P450 isozymes. Arch Biochem Biophys 2002;405(2):223-30.
37. Gupta SC, Hevia D, Patchva S, Park B, Koh W, Aggarwal BB. Upsides and downsides of reactive oxygen species for cancer: The roles of reactive oxygen species in tumorigenesis, prevention, and therapy. Antioxid Redox Signal 2012;16(11):1295-322.
38. Dance-Barnes ST, Kock ND, Moore JE, Lin EY, Mosley LJ, Dâ€™gostino RB Jr, et al. Lung tumor promotion by curcumin. Carcinogenesis 2009;30(6):1016-23.
39. Berridge MV, Herst PM, Tan AS. Tetrazolium dyes as tools in cell biology: New insights into their cellular reduction. Biotechnol Annu Rev 2005;11:127-52.
40. Amruthraj NJ, Raj JP, Saravanan S, Lebel A. In vitro studies on anticancer activity of capsaicinoids from Capsicum Chinese against human hepatocellular carcinoma cells. Int J Pharm Pharm Sci 2014;6(4):254-8.
41. Barltrop J. 5-(3-carboxymethoxyphenyl)-2-(4,5-dimenthylthiazoly)-3-(4-sulfophenyl)tetrazolium inner salt (MTS) and MTS analogs of 3-(4,5-dimethylthiazolyl)-2,5-diphenyltetrazolium bromide (MTT) reducing to purple water-soluble formazans as cell-viability indicators. Bioorg Med Chem Lett 1991;1:611-4.
42. Sakagami H, Kobayashi M, Chien CH, Kanegae H, Kawase M. Selective toxicity and type of cell death induced by various natural and synthetic compounds in oral squamous cell carcinoma. In Vivo 2007;21(2):311-20.
The publication is licensed under CC By and is open access. Copyright is with author and allowed to retain publishing rights without restrictions.