Cytotoxic and Antiproliferation Activity of n-Hexane Fraction of Avocado seed (Persea americana Mill.) on MCF7 cell

Juwita Rahwawati(1*), Maryati Maryati(2), Ratna Yuliani(3)

(1) Universitas Muhammadiyah Surakarta
(2) Universitas Muhammadiyah Surakarta
(3) Coventry University
(*) Corresponding Author

Abstract

Avocado seeds, which often end up as food waste, have many health benefits, including anticancer activity. This study aimed to determine the potential cytotoxic activity of avocado seeds against breast cancer cells, MCF7. Avocado seeds were macerated with 96% ethanol and fractionated with n-hexane, chloroform, and ethyl acetate as solvents. Cytotoxic activity of the extract and fractions was tested using the MTT method, while antiproliferation activity and apoptosis of the n-hexane fraction were tested using the doubling time and double staining methods, respectively. The selectivity of the n-hexane fraction was also determined. The results showed that the extract and n-hexane fraction of avocado seeds had cytotoxic activity on MCF7 with IC50 values of 211.2±25 and 47.87±1.5 µg/mL, respectively. The cytotoxic activity of the n-hexane fraction was selective (Selectivity Index = 4.73). The fraction can extend the doubling time to 76.54-77.41 hours. The double staining test results showed red fluorescence in the cells, indicating the apoptosis of MCF7 cells. The n-hexane fraction of avocado seed extract has the potential to be developed as an anticancer therapeutic agent.

 

Keywords

Persea americana Mill., MCF7, MTT, cytotoxic, apoptosis

Full Text:

PDF

References

Alkhalaf, M. I., Alansari, W. S., Ahmed, E., & Elhalwagy, M. E. A., 2019. Anti-oxidant, anti-inflammatory and anti-cancer activities of avocado (Persea americana) fruit and seed extract: Journal of King Saud University - Science, 31, pp.1358–1362.

Ammerman, N. C., Beier-Sexton, M., & Azad, A. F., 2009. Growth and Maintenance of Vero Cell Lines Nicole: NIH, pp.1–10.

Bjelaković, G., Nagorni, A., Bjelaković, M., Stamenković, I., Arsic, R., & Katic, V., 2005. Apoptosis: Programmed Cell Death and Its Clinical Implications: Medicines and Biology, 12(1), pp.6–11.

Carvalho, C., Santos, R. X., Cardoso, S., Correia, S., Oliveira, P. J., Santos, M. S., & Moreira, P. I., 2009. Doxorubicin : The Good, the Bad and the Ugly Effect Doxorubicin: Current Medicinal Therapy, pp.16-25.

CCRC., 2009a. Prosedur Tetap Pengamatan Apoptosis dengan Metode Double Staining : (CCRC-03-011-01).

CCRC., 2009b. Prosedur Tetap Uji Pengamatan Proliferasi Sel (Doubling Time): (CCRC-03-015-01).

CCRC., 2012. Prosedur Tetap Uji Sitotoksik Metode MTT: (CCRC-03-010-01).

Dabas, D., Elias, R. J., Ziegler, G. R., & Lambert, J. D., 2019. In Vitro Antioxidant and Cancer Inhibitory Activity of a Coloured Avocado Seed Extract: International Journal of Food Science, pp.1-7.

Dabas, D., Shegog, R., Ziegler, G., & Lambert, J., 2013. Avocado (Persea americana) Seed as a Source of Bioactive Phytochemicals: Current Pharmaceutical Design., pp.19-34.

D’Ambrosio, S. M., Han, C., Pan, L., Kinghorn, A. D., & Ding, H., 2011. Aliphatic Acetogenin Constituents of Avocado Fruits Inhibit Human Oral Cancer Cell Proliferation by Targeting the EGFR/RAS/RAF/MEK/ERK1/2 Pathway: National Institute of Health, 10, 409(3), pp.1–11.

Dewi, A., Halimah, N., Rohmah, S. S., Fakultas, M., Masyarakat, K., Diponegoro, U., & Indonesia, S., 2014. Pengolahan Limbah Biji Alpukat Untuk Pembuatan Dodol Pati Sebagai Alternatif Pengobatan Ginjal: Jurnal Ilmiah Mahasiswa, 4, pp.32–37.

Edem, D. O., Ekanem, I. S., & Ebong, P. E., 2009. Persea americana Mill on Blood Glucose and Histopathology of Pancreas in Alloxan-Induced Diabetic Rats: Journal of Diabetes Mellitus 22(3), pp.272–276.

Foglieni, C., Meoni, C., & Davalli, A. M., 2001. Fluorescent Dyes for Cell Viability, An Application on Prefixed Conditions: Springer, 115, pp.223–229.

Fulgoni, V. L., Dreher, M., & Davenport, A. J., 2013. Avocado Consumption is Associated with Better Diet Quality and Nutrient Intake, and Lower Metabolic Syndrome Risk in US Adults : Results from The National Health and Nutrition Examination Survey (NHANES) 2001 – 2008: Nutrition Journal, 12:1, pp.1–6.

Grace, S. S. R., Chauhan, J. B., & Jain, C. R., 2015. Preliminary Phytochemical Investigation and TLC Analysis of Peel and Seed Extracts of Persea Americana: Asian Journal of Pharmaceutical Science & Technology, 5(3), pp.167–171.

Kristanty, R. E., Suriawati, J., & Sulistyo, J., 2014. Cytotoxic Activity of Avocado Seeds Extracts (Persea americana Mill.) on T47D Cell Lines: International Research Journal of Pharmacy, 5(7), pp.557–559.

Lara-Márquez, M., Báez-Magaña, M., Raymundo-Ramos, C., Spagnuolo, P. A., Macías-Rodríguez, L., Salgado-Garciglia, R., Ochoa-Zarzosa, A., & López-Meza, J. E., 2020. Lipid-rich extract from Mexican avocado (Persea americana var. drymifolia) induces apoptosis and modulates the inflammatory response in Caco-2 human colon cancer cells: Journal of Functional Foods, pp. 1-10.

Malumbres, M., & Barbacid, M., 2007. Cell Cycle Kinases in Cancer: Genome Biology, 17, pp.60–65.

Mardiyaningsih, A., & Ismiyati, N., 2014. Cytotoxic Activity of Ethanolic Extract of Persea americana Mill. Leaves On HeLa Cervical Cancer Cell: Traditional Medicine Journal, 19, pp.24–28.

Maryati, M., Sutrisna, E., Saifudin, A., Kusumaningrum, I. D., & Abu bakar, M. F., 2019. Extract of Ceplukan (Physalis angulata L.) Inhibited Proliferation and Induced Apoptosis in Myeloma Cell Line: Food Research, pp.755–760.

McCormack, L. A., Laska, M. N., Larson, N. I., & Story, M., 2010. Review of The Nutritional Implications of Farmers’ Markets and Community Gardens, A Call for Evaluation and Research Efforts: Journal of the American Dietetic Association, 110(3), pp.399–408.

Mehrara, E., Forssell-Aronsson, E., Ahlman, H., & Bernhardt, P., 2007. Specific Growth Rate Versus Doubling Time for Quantitative Characterization of Tumour Growth Rate: Cancer Research, 67(8), pp.3970–3975.

Nur, A., Abubakar, F., Achmadi, S. S., & Suparto, I. H., 2017. Triterpenoid of avocado (Persea americana) seed and its cytotoxic activity toward breast MCF-7 and liver HepG2 cancer cells: Asian Pacific Journal of Tropical Biomedicine, 7(5), pp.397–400.

Nwaoguikpe, R. N., 2011. The Effect of Aqueous Seed Extract of Persea Americana (Avocado Pear) on Serum Lipid and Cholesterol Levels in Rabbits: International Research Journal in Pharmacy, 1, pp.23-39.

Pahua-ramos, M. E., Ortiz-moreno, A., Necoechea-mondragón, H., & Hernández-ortega, M., 2012. Hypolipidemic Effect of Avocado (Persea americana Mill) Seed in a Hypercholesterolemic Mouse Model: Plant Foods Hum Nutr, 67., pp.10–16.

Rahmawati, J., & Maryati, M., 2021. Aktivitas Sitotoksik dan Antiproliferasi Fraksi n-Heksan Biji Al-pukat (Persea americana Mill.) Terhadap sel T47D. Pharmacon: Jurnal Farmasi Indonesia, 18(1), pp.38–46.

Rivai, H., Andalas, U., & Putri, Y. T., 2019. Qualitative and Quantitative Analysis of the Chemical Content of Hexane, Acetone Ethanol and Water Extract from Avocado Seeds (Persea americana): Scholars Middle East Publisher, pp.24–31.

Shehata, M. M. S. M., & Solttan, S. S. A., 2013. Effects of Bioactive Component of Kiwi Fruit and Avocado (Fruit and Seed) on Hypercholesterolemic Rats: World Journal of Dairy & Food Sciences, 8(1), pp.82–93.

Sutejo, I. R., Putri, H., & Meiyanto, E., 2016. Selektivitas Ekstrak Etanolik Buah Makassar (Brucea javanica) pada Kanker Payudara Metastasis secara In Vitro: Journal of Agromedicine and Medical Sciences, 2(1), pp.1–5.

Velez-pardo, C., 2014. Pro-apoptotic effect of Persea americana var. Hass (avocado) on Jurkat lymphoblastic Leukaemia Cells: Informa Healthcare, 52(4), pp.458–465.

Vo, T. S., Le, P. U., & Ngo, D. H. (2019). Free radical scavenging and anti-proliferative activities of avocado (Persea americana Mill.) seed extract: Asian Pacific Journal of Tropical Biomedicine, 9(3), pp.91–97.

Wati, E. M., Puspaningtyas, A. R., & Pangaribowo, D. A., 2016. Uji Sitotoksisitas dan Proliferasi Senyawa 1- (4-nitrobenzoiloksi- metil) -5-fluorourasil terhadap Sel Kanker Payudara MCF-7: Pustaka Kesehatan, 4(3), pp.484–488.

WCRF., 2018. Breast Cancer Statistics World Cancer Research Fund: World Cancer Research Fund., https://www.wcrf.org/dietandcancer/cancer-trends/breast-cancer-statistics.

Widiastuti, Y., Pratiwi, R., Riyanto, S., & Wahyuono, S., 2018. Persea americana Mill. Seed Extract on MCF-7 Cancer Cell Line. Indonesian Journal of Biotechnology: 23(2), pp.61–67.

Wong, R. S., 2011. Apoptosis in cancer; from pathogenesis to treatment: Journal of Experimental & Clinical Cancer Research, 30:87(2), pp.1–14.

Yahia, E. M., 2012. Avocado: Crop Post-Harvest, Science and Technology, 1, pp.159-186.

DOI: 10.23917/pharmacon.v19i1.18274

Article Metrics

Abstract view(s): 341 time(s)
PDF: 310 time(s)

Refbacks

  • There are currently no refbacks.