Isaac Scientific Publishing

Journal of Advances in Molecular Biology

Phytochemical Profiling and Anti-cancer Study of Lyophilized Pure Fruit Juice of Citrus Limon (L.) Osbeck against Human Breast Cancer (MCF-7) Cell Line

Download PDF (506.3 KB) PP. 91 - 104 Pub. Date: September 1, 2017

DOI: 10.22606/jamb.2017.12002

Author(s)

  • Saranya. A.
    PG and Research Department of Zoology Presidency College (Autonomous) Chennai-5
  • Sivakumari. K.*
    Associate Professor of Zoology, Presidency College, Chennai.
  • Ashok. K.
    PG and Research Department of Zoology Presidency College (Autonomous) Chennai-5
  • Rajesh. S
    PG and Research Department of Zoology Presidency College (Autonomous) Chennai-5

Abstract

The present study aims at the phytochemical profiling of lyophilized pure fruit juice powder of Citrus limon and demonstrates its efficacy as an antitumor agent against human breast cancer cell line (MCF-7). The cytotoxicity was evaluated using the changes in cell morphology, cell viability, and DNA fragmentation, the percentage of cell viability was determined by MTT assay. Our results showed that lyophilized pure fruit juice powder of Citrus limon inhibited the proliferation of human breast cancer cell line MCF-7 with an IC50 98.16 μg/ml at 48 h incubation, it was shown to promote apoptosis as seen as DNA fragmentation using ladder assay. These results suggest that lyophilized pure fruit juice powder of Citrus limon has antiproliferative effect against MCF-7 cell by suppressing its growth.

Keywords

Citrus limon fruits, FT-IR, GC-MS, MTT, DNA fragmentation.

References

[1] Alison, MR., (2001). Department of Histopathology and hepatology Imperial college school of medicine London and histopathology unit. Imperial cancer Research Fund. London, UK Liver., 21: 367-73.

[2] Jemal, A., Murray, T., Samuels, A., Ghafoor, A., Ward, E. and Thun, M. (2003). Cancer statistics. Cancer J. Clin., 53: 5‐26.

[3] Balachandran, P. and Govindarajan, R. (2005). Cancer an ayurvedic perspective. Pharmaco. Res., 51: 9-30.

[4] Siegel, RL., Miller, KD. and Jemal, A. (2015). Cancer statistics. CA Cancer J. Clin., 65(1): 5-29.

[5] Kumbhare, MR., Dadmal, TL., Ramaiah, JM., Kishore, VSK., Valli, P., Tiwari, KM., Appalanaidu, K., Rao, KY. and Bhadra, PM. (2015). Synthesis and anticancer of novel triazole linked N-(pyrimidin-2-yl)benzol[d]thaizol-2-amine derivatives as inhibitors of cell survival proteins and inducers of apoptosis in MCF-7 breast cancer cells. Bioorg. Med. Chem. Lett., 25: 654-658.

[6] Tavakoli, J., Miar, S., Zadehzare, MM. and Akbari, H. (2012). Evaluation of effectiveness of herbal medication in cancer care: A Review Study. Iran. J. Cancer Prevent., 5(3): 144-56.

[7] American Cancer Society. (2015). For written permission, address the Legal department of the American Cancer Society. 250 Williams Street, NW, Atlanta, GA: 30303-1002.

[8] Ferlay, JBF., Pisani, P. and Parkin, DM. (2000). GLOBOCAN 2000. Cancer incidence and mortality and prevalence worldwide. Version 1.0. IARC Cancer., 19: 1941-52.

[9] Ferlay, J., Shin, HR., Bray, F., Forman, D., Mathers, C. and Parkin, DM. (2010). Estimates of worldwide burden of cancer in 2008: GLOBOCAN 2008. Int. J. Can., 127(12): 2893–2917.

[10] Harris, JR., Lippman, ME., Morrow, M. and Osborne, CK. (2000). Treatment of metastatic breast cancer, in: J.R. Harris, M.E. Lippmann, M. Morrow (Eds.), Diseases of the Breast, 2nd ed., Lippincott Williams and Wilkins, Philadelphia, 749.

[11] Cancer Research UK. (2013). Side Effects of Cancer Drugs. Available online: http://cancerhelp. cancerresearchuk.org/about cancer treatment cancer drugs side-effects.

[12] Raguz, S. and Yague, E. (2008). Resistance to chemotherapy: New treatments and novel insights into an old problem. Br. J. Cancer., 99: 387-391.

[13] Jemal, A., Siegel, R., Ward, E., Murray, T., Xu, J. and Thun, MJ. (2007). Cancer Statistics. CA Cancer J. Clin., 57: 43-66.

[14] Anand, P., Kunnumakara, AB., Sundaram, C., Harikumar, KB., Tharakan ST., Lai, OS, Sung, B. and Aggarwal, BB. (2008). Pharm. Res., 25: 2097.

[15] Sun, Y., Xun, K., Wang, Y. and Chen, X. (2009). The effects of plant falavonoids on mammalians cells. Anticancer. Drugs., 20: 757-769.

[16] Dev, S. (2000). Impact of natural products in modern drug development. Ind. J. Exp. Biol., 48: 191-198.

[17] Srivastava, SK., Rai, V., Srivastava, M., Rawat, AKS. and Mehrotra, S. (2000). Estimation of heavy metals in different berberis species and its market samples. Environ. Monit. Assess., 116: 315-320.

[18] Newman, DJ., Cragg, GM. and Snader, KM. (2003). Natural products as sources of new drugs over the period. J. Nat. Prod., 66: 1022-1037.

[19] Yang, HL., Chen, CS., Chang, WH., Lu, FJ., Lai, YC. and Chen, CC. (2006). Growth inhibition and induction of apoptosis in MCF-7 Breast cancer cells by Antodia camohorata. Cancer. Lett., 231(2): 215-27.

[20] Gupta, SC., Kim, JH., Prasad, S. and Aggarwal, BB. (2010). Regulation of survival, proliferation, invasion, angiogenesis, and metastasis of tumor cells through modulation of inflammatory pathways by nutraceuticals, Cancer Metastasis Rev., 29: 405-434.

[21] Agarwal, N., Majee, C. and Chakraborthy, GS. (2012). Natural herbs as anticancer drugs. Int. J. Pharam. Tech. Res., 4(3): 1142-1153.

[22] Gopinath, P., Vadivel, SD., Kamatchiammal, S. and Saroja, V. (2013). Anticancerous activity of Albizia amara (Roxb.) Bovin using human breast cancer cells (MCF-7) by in vitro methods. Int. J. Pharma. Res. Rev., 2(8): 23-32.

[23] Garcia Sancho, LEG., Yahia, EM., Garcia-Solis, P. and Gonzalez-Aguilar, GA. (2014). Inhibition of proliferation of breast cancer cells MCF-7 and MDA-MB-231 by lipophilic extracts of Papaya (Carica papaya L. var. Maradol) Fruit. Food. Nutr. Sci., 5: 2097-2103.

[24] Kalla, PK., Chitti, S., Aghamirzaei, ST., Senthilkumar, R. and Selvam, A. (2014). Anticancer activity ossilymarin on MCF-7 and NCIH-23 cell lines. Ad. J. Bio. Res., 8(2): 57-61.

[25] Graidist, P., Martla, M. and Sukpondma, Y. (2015). Cytotoxic activity of piper extracts in breast cancer cell lines. Nutrients., 7: 2707-2718.

[26] Serasanambati, M., Chilakapati, SR., Manikonda, PK. and Kanala, JR. (2015). Anticancer activity of methanolic extract of berberis in MCF-7 human breast cancer cell lines. Int. J. Life Sci. Biotech. Pharma. Res., 4(1): 443-461.

[27] Venkateswarlu, G., Rani Swarupa, M., Vani, PA. and Vineela, J. (2015). In vitro anticancer activity of petroleum ether extract of Cynodon dactylon. J. Pharma. Phytochem., 4(1): 164-168.

[28] Arora, R., Agarwal, P., Ray, S., Singh, AK., Singh, VJ., Tyagi, AK. and Kapoor, S. (2007). MADS-box gene family in rice genome-wide identification, organization and expression profiling during reproductive development and stress. BMC Genomics., 8: 242(1-21).

[29] Goktepe, M. and Ahmedna, L. (2012). Maness In vitro cancer research on ancient herbal remedies: A changing trend. J. App. Pharma. sci., 2(8):13-20.

[30] Rekha, S. and Bhaskar, M. (2013). In vitro screening and identification of antioxidant activities of orange (Citrus sinesis) peel extract in different solvents. Int. J. pharm. Bio. Sci., 4(4): 405-412.

[31] Adnan, M., Umer, A., Ahamed, I., Hayat, K. and Shakeel, NS. (2014). In vitro evalution of biological activities of citrus leaf extracts. Sains Malaysiana., 43(2):185-194.

[32] Parashar, S., Sharma, H. and Garg, M. (2014). Antimicrobial and antioxidant activities of fruit and vegetable peels: A review. J. Pharmacol. Phytochem., 3(1): 160-164.

[33] Safarzadeh, E., Shotorbani, SS. and Baradaran, B. (2014). Herbal medicine as inducers of apoptosis in cancer treatment. Adv. Pharm. Bull., 4(1): 421-427.

[34] Singh, S. and Singh, R. (2014). Herbal medicine treatment of malaria in aliero local government area, kebbi, Nigeria. J. M. Plants Stu., 2(2): 117-126.

[35] Mehra, S., Shukla, S., Srivastava, R., Mathew, J. and Mehra, M. (2015). Evalution of antimicrobial activity of peel and pulp extracts of C. paradise, C.medica and Limon against B. Cereus and M. Luteus. Australian. J. Basic. Applied. Sci., 9(1): 174-182.

[36] Rosangkima, G. and Prasad, SB. (2004). Antitumour activity of some plants from Meghalaya and Mizoram against murine ascites Dolton's lymphoma. Indian J. Exp. Biol., 42: 981-988.

[37] Chong, FW., Srikumar, C., Nagaraja, HS., Thanikachalam, PM. and Nagarajah, L. (2009). Expression of transforming growth factor-β and determination of Apoptotic Index in histo pathological sections for assessment of the effects of Apigenin (4',5',7'-trihydroxyflavone) on Cyclosporine A induced renal damage. Malaysian J. Path., 31(1): 35-43.

[38] Al-anbari, AK., Alkhesraji, T., Al-Mashhadani, A. and Aljewari, H. (2013). Determination of some chemical compounds in Citrus species in Iraq. Int. J. Ad. Res., 1(7): 555-562.

[39] Duyn, MA. and Pivonka, E. (2000). Overview of the health benefits of fruits and vegetable consumption for the dietetics professional. J. Amer. Diet Association., 100(2): 1511-1521.

[40] Dhanavade, MJ., Jalkute, CB., Ghosh, JS. and Sonawane, KD. (2011). Study Antimicrobial Activity of Lemon (Citrus lemon L.) Peel Extract. British. J. Pharmacol. Toxicol., 2(3): 119-122.

[41] Sawalha, SMS., Arraez-Roman, D., Segura-Carretero, A. and Fenandez-Gutierrez, A. (2009). Quantification of main phenolic compounds in sweet and bitter orange peel using CEMS/MS. Food. Chem., 116: 567-574.

[42] Singh, S. and Rajam, MV. (2009). Citrus biotechnology: Achievements, limitations and future directions. Physio. Mol. Bio. Plants., 15(1): 3-22.

[43] Ladaniya, MS. (2008). Citrus Fruit: Biology, technology and evaluation. Int. J. Phama. Sci., 12: 634-642.

[44] Carina, LA., Ingrid, WL., Isabelle, CD., Tonia, VSL., Adeliana, SO., Maria, RAM., Edda, LL. and Mauricio, PS. (2004). Biological activity of proteins from pulps of tropical fruits. Food Chem., 85: 107-110.

[45] Kokate, CK. (1988). Practical pharmacognosy, 2nd edition, Vallabh prakashan publishers, New Delhi., 4-29.

[46] Mosmann T (1983). Rapid colorimetric assay for cellular growth and survival: application to proliferation and cytotoxicity assays. J.Immunol.Methods, 65: 55-63.

[47] Yawata, A. (1998). Prolonged cell survival enhances peritoneal dissemination of gastric cancer cells. Oncogene., 16: 2681-6.

[48] Merina, N., Chandra, KJ. and Jibon, K. (2012). Medicinal plants with anticancer activities: A rev., 3(6): 2230-8407.

[49] Larkin, T. (1983). Herbs are often more toxic than magical. FDA. Consum., 17: 4-11.

[50] Saxe, TG. (1987). Toxicity of medicinal herbal preparations. Am. Fam. Physician., 35: 135-42.

[51] Mishra, LC., Singh, BB. and Dagenais, S. (2000). Basis for the Therapeutic use of Withania somnifera (Ashwagandha): A Review. Alter. Medi. Rev., 5(4): 334-336.

[52] Zhang, YJ., Abe, T., Tanaka, T., Yang, CR. and Kouno, I. (2001). Phyllan emblinins A-F, new Ellagitannins from Phyllanthus emblica. J. Nat. Prod., 64: 1527-1532.

[53] Cragg, GM. and Newman, DJ. (2005). plants as a source of anticancer agents. J. Ethnopatho., 100: 72-79.

[54] Kumar, VL. and Arya, S. (2006). Medicinal uses and pharmacological properties of Calotropis procera. In recent progress in medicinal plants., 11: 378-388.

[55] Ahamed-Belkacem, A., Macalou, S., Borrelli, F., Capasso, R., Fattorusso, E. and Taglialatela-Scafati, O. (2007). Nonprenylated rotenoids, a new class of potent breast cancer resistance protein inhibitors. J. Med. Chem., 50(8): 1933-1938.

[56] Norikura, T., Kojima-Yuasa, A., Shimizu, M., Huang, X., Xu, S. and Kametani, S. (2008). Mechanism of growth inhibitory effect of Blumea Balsamifera extract in Hepatocellular Carcinoma. Biosci. Biotech. Biochem., 72(5): 1183-1189.

[57] Kundusen, S., Gupta, M., Mazumder, UK., Haldar, PK., Saha, P. and Bala, A. (2011). Antitumor activity of Citrus maxima (burm.) Merr. leaves in ehrlichs ascites carcinoma cell-treated mice. ISRN. Parmacol., 2: 1-8.

[58] Nair, S. and Varalakshmi, KN. (2011). Anticancer, cytotoxic potential of Morninga olifera extracts on HeLa cell lines. J. Natur. Pharm., 2(3): 138-142.

[59] Seawan, N., Koysomboon, S. and Chantrapromma, K. (2011). Anti-trypsinase and anticancer activities of flavonoids from Blumea Balsamifera Dc. J. Med. Plants. Res., 5(6): 1018-1025.

[60] Hashim, MN., Rahmari, M., Cheng Lian, EG., Sukari, MA. and Yahayu, M. (2012). Antiproliferative Activity of Xanthones isolated from Artocarpus obtusus. Journal of biomedicine and Biotechnology. 02: 681-685.

[61] Prabhu, D., Arulvasu, C., Babu, G., Manikandan, R. and Srinivasan, P. (2013). Biologically synthesized green silver nanoparticles from leaf extract of Vitex negundo L. induce growth-inhibitory effect on human colon canver cell line HCT15. Process Biochem., 48: 317-324.

[62] Ashok, K. and Sivakumari, K. (2015). In silico docking of fucoidan compound against the selective proteins of HepG-2 cell line. Int. J. Chem. Pharm. Sci., 6(4): 976-979.

[63] Manimaran, M., Sivakumari, K. and Ashok, K. (2015). Molecular studies of reservatrol against the human oral cancer cell line proteins (KB cells). Int. J. curr. Adv. Res., 4: 2319-2325.

[64] Muthukala, B., Sivakumari, K. and Ashok, K. (2015). In silico docking of quercetin compound against the hela cell line proteins. Int. J. Curr. Pharm. Res., 7(1): 0975-0980.

[65] Manimekalai, I., Sivakumari, K., Ashok, K. and Rajesh, S. (2016). Phytochemical profiling of mangosteen fruit, Garcinia mangostana. World. J. Pharm. Sci., 5(2): 221-252.

[66] Manimekalai, I., Sivakumari, K., Ashok, K. and Rajesh, S. (2016). Antioxidant and anticancer potential of mangosteen fruit, Garinia mangostana against hepatocellular carcinoma (HepG-2) cell line. World. J. Pharm. Sci., 5(2): 253-293.

[67] Rajesh, S., Sivakumari, K., Ashok, K. and Abitha, AR. (2016). Anticancer activity of Cardiospermum helicacacabum leaf extracts against hepatocellular carcinoma cell line (HepG-2). World. J. Pharm. Sci., 5(3): 1133-1154.

[68] Waterman, PG. and Grundon, MF. (1983). Chemistry and chemical taxonomy of the

[69] Mohanapriya, M., Ramaswamy, L. and Rajendren, R. (2013). Health and medicinal properties of lemon (Citrus Limonum). Int. J. Ayur. Herb. Med., 3(1): 1095-1100.

[70] Kegley, SE., Hill, BR., Orme, S. and Choi, AH. (2005). Limonene pan pesticide database. Ind. Chem., 8(1): 572-578.

[71] Chede, PS. (2013). Phytochemical analysis of citrus sinesis peel. Int. J. Pharm. Bio. Sci., 4(1): 339-343.

[72] Burt, SA. (2004). Essential oils: their antibacterial properties and potential applications in foods: A review, Int. J. Food. Microbial., 24: 223-253.

[73] Ortuno, A., Gomez, P., Baidez, A., Frias, V. and Del Rio, JA. (2006). Citrus sp: a source of flavonoids of pharmaceutical interest potential health benefits of Citrus. ACS Symp. Ser., 936: 175-185.

[74] Strange, RR., Miland, SL., Eckert, JW. and Sims, JJ. (1993). An antifungal compound produced by grapefruit and valencia orange after wounding of the peel, J. Nat. Prod., 56: 1627-1629.

[75] Crowell, PL. (1999). Prevention and therapy of cancer by dietary monoterpenes. The J. Nutr., 129(3): 775S-778S.

[76] Tsuda, H., Ohshima, Y. and Nomoto, H. (2004). Cancer prevention by natural compound. Drug Metab. Pharm., 19(4): 245-63.

[77] Shahnah, SM., Ali, S., Ansari, H. and Bagri, P. (2007). New sequiterpene derivative from fruit peel of Citrus limon (Linn) Burn. Food Sci. Pharm., 75: 165-170.

[78] Ahmad, MM., Salim-ur-Rehman, Z., Iqbal, FM., Anjum, N. and Sultan JI. (2006). Genetic variability to essential oil composition in four citrus fruit species. Pak. J. Bot., 38(2): 319-324.

[79] Aronson, JK. (2001). Nature Publishing Group. Retrieved from: http://medicine.nature.com.

[80] Ighodaro, O. (2012). Evaluation study on Nigerian species of Musa paradisiaca peels: phytochemical screening, proximate analysis, mineral composition and antimicrobial activities. Res., 4: 17-20.

[81] El Zawawy, AN. (2015). Antioxidant, antitumor, antimicrobial studies and quantitative phytochemical estimation of ethanolic extracts of selected fruit peels. Int. J. Curr. Microbiol. App. Sci., 4(5): 298-309.

[82] Hashemi, M., Karami-Tehrani, F. and Ghavami, S. (2004). Cytotoxicity effect of Cladribine on the MCF-7 Human breast cancer cell line. Iran. Biomed. J., 8(1): 7-12.

[83] Yang, HL., Chen, CS., Chang, WH., Lu, FJ., Lai, YC. and Chen, CC. (2006). Growth inhibition and induction of apoptosis in MCF-7 Breast cancer cells by Antodia camohorata. Cancer. Lett., 231(2): 215-27.

[84] Khalil, AM. (2009). Inhibition of the vitro growth of human mammary carcinoma cell line (MCF-7) by selenium and vitamin E. J. Bio. Sci., 2: 37-46.

[85] Ibrahim, T., Mercatali, L., Sacanna, E., Tesei, A., Carloni, S., Ulivi, P., Liverani, C., Fabbri, F., Zanoni, M., Zoli, W. and Dino, A. (2012). Inhibition of breast cancer cell proliferation in repeated and non-repeated treatment with Zoledronic acid. Cancer Cell Int., 12: 48(12).

[86] AbuHammad, S. and Zihlif, M. (2013). Gene expression alterations in doxorubicin resistant MCF7 breast cancer cell line. Genomics., 101: 213-220.

[87] Sadowska, A., Prokopiuk, S., Miltyk, W., Surazynski, A., Konoczuk, J., Sawicka, D. and Car, H. (2013). Metronidazole affects breast cancer cell lines. Anticancer drugs., 58(1): 90-95.

[88] Wu, S., Guo, J., Wei, W., Zhang, J., Fang, J. and Beebe, JS. (2014). Enhanced breast cancer therapy with nsPEFs and low concentrations of gemcitabine. Cancer Res., 14: 98.

[89] Swapana, M. and Padamavathy, C. (2015). A critical review on breast cancer literature: screening awareness and preventive measures. Mediterranean. J. Soc. Sci., 6(4): 2039-2117.