The Effect of Nutmeg Seed (M. fragrans) Extracts Induces Apoptosis in Melanoma Maligna Cell’s (B16-F10)

Susianti Susianti, Ronny Lesmana, Supriatno Salam, Euis Julaeha, Yuni Susanti Pratiwi, Nova Sylviana, Hanna Goenawan, Ahmad Kurniawan, Unang Supratman


BACKGROUND: Nutmeg (Myristica fragrans H.), one of native plants of Maluku Indonesia, has long been used as traditional medicines especially to treat tumors, externally to treat skin infections. M. fragrans also has important biological activities as anticancer. However, antimelanoma activity of M. fragrans remains unknown. The aim of this study is to compare M. fragrans extracts as anticancer on melanoma B16-F10 cells by inducing apoptosis.

METHODS: M. fragrans seed was extracted with ethanol then fractionated with n-hexane, ethyl acetate, and n-butanol. B16-F10 melanoma cells were cultured and treated with various doses and tested using resazurin reduction assay. Apoptosis signalling via caspase-3 was measured by using western blot.

RESULTS: The extract and fractions of M. fragrans reduced viability of cells with IC50 value for ethanol extract 21.83 µg/mL, ethyl acetate fraction 21.66 µg/mL, n-hexane fraction 47.53 µg/mL, and n-butanol fraction >1,000 µg/mL. The active fraction of ethyl acetate induced apoptosis via caspase-3 proteins similar with cisplatin as positive control in B16-F10 cells at 10 hours treatment.

CONCLUSION: Taken together, M. fragrans ethyl acetate fraction has the highest IC50 than n-hexane and n-butanol fractions that significantly inhibited B16-F10 cell proliferation by inducing apoptosis via caspase-3. It provides the insight that it has the most potential activity as a chemopreventive agent for addressing melanoma skin cancer.

KEYWORDS: M. fragrans, apoptosis, fraction, melanoma

Full Text:



Zaidi MR, Fisher DE, Rizos H. Biology of melanocytes and primary melanoma. In: Balch C, Editor. Cutaneous Melanoma. Cham: Springer; 2020. p.3-40, CrossRef.

Linares MA, Zakaria A, Nizran P. Skin cancer. Prim Care. 2015; 42: 645-59, CrossRef.

Davis EJ, Perez MC, Ayoubi N, Zhao S, Ye F, Wang DY, et al. Clinical correlates of response to Anti-PD-1–based therapy in patients with metastatic melanoma. J Immunother. 2019; 42: 221-7, CrossRef.

World Health Organization [Internet]. Radiation: Ultraviolet (UV) radiation and skin cancer. [updated 2017 Oct 16; cited: 2019 April 1]. Available from:

Alvarez-Artime A, Cernuda-Cernuda R, Francisco-Artime-Naveda, Cepas V, Gonzalez-Menendez P, Fernadez-Vega S, et al. Melatonin-induced cytoskeleton reorganization leads to inhibition of melanoma cancer cell proliferation. Int J Mol Sci. 2020; 21: 548, CrossRef.

Carr S, Smith, C, Wernberg, J. Epidemiology and risk factors of melanoma. Surg Clin North Am. 2020; 100: 1-12, CrossRef.

Tracey EH, Vij A. Updates in melanoma. Dermatol Clin. 2019; 37: 73-82, CrossRef.

Yu Q, Xie J, Li J, Lu Y, Liao L. Clinical outcomes of BRAF plus MEK inhibition in melanoma: A meta‐analysis and systematic review. Cancer Med. 2019; 8: 5414-24, CrossRef.

Domingues B, Lopes J, Soares P, Populo H. Melanoma treatment in review. Immunotargets Ther. 2018; 7: 35-49, CrossRef.

Lesmana R, Susianti, Pediatama T, Sylviana N, Pratiwi YS, Goenawan H, et al. Active compounds from curcuma longa and comparison of their effectively induced apoptosis in MCF-7 cell. Pak J Biol Sci. 2020; 23: 35-41, CrossRef.

Rodianawati I, Hastuti P, Cahyanto MN. Nutmeg’s (Myristica fragrans Houtt) oleoresin: effect of heating to chemical compositions and antifungal properties. Procedia Food Sci. 2015; 3: 244-54, CrossRef.

Gupta AD, Bansal VK, Babu V, Maithil N. Chemistry, antioxidant and antimicrobial potential of nutmeg (Myristica fragrans Houtt). J Genet Eng Biotechnol. 2013; 11: 25-31, CrossRef.

Abourashed EA, El-Alfy AT. Chemical diversity and pharmacological significance of the secondary metabolites of nutmeg (Myristica fragrans Houtt.). Phytochem Rev. 2016; 15: 1035-56, CrossRef.

Kuete V. Medicinal spices and vegetables from Africa. 1st ed. Cambridge: Academic Press; 2017, NLMID.

Le TVT, Nguyen PH, Choi HS, Yang JL, Kang KW, Ahn SG, et al. Diarylbutane-type lignans from Myristica fragrans (Nutmeg) show the cytotoxicity against breast cancer cells through activation of AMP-activated protein kinase. Nat Prod Sci. 2017; 23: 21-8, CrossRef.

Matulyte I, Jekabsone A, Jankauskaite L, Zavistanaviciute P, Sakiene V, Bartkiene E, et al. The essential oil and hydrolats from Myristica fragrans seeds with magnesium aluminometasilicate as excipient: antioxidant, antibacterial, and anti-inflammatory activity. Foods. 2020; 9: 1-12, CrossRef.

Mishra A, Rahman S, Khan R. CNS activity of Myristica fragrans Houtt. - An experimental study. Bangladesh J Med Sci. 2018; 17: 98-106, CrossRef.

Lestari K, Diantini A, Barliana MI, Achmad TH, Subarnas A, Mutaki, et al. Potential natural dual agonist PPARa/y-induced antidiabetic and antidyslipidemic properties of safrole-free nutmeg seed (Myristica fragrans Houtt) extract. Nat Prod J. 2019; 9: 248-53, CrossRef.

Domura R, Sasaki R, Ishikawa Y, Okamoto M. Cellular morphology-mediated proliferation and drug sensitivity of breast cancer cells. J Funct Biomater. 2017; 8: 18, CrossRef.

Chaouki W, Leger DY, Eljastimi J, Beneytout JL, Hmamouchi M. Antiproliferative effect of extracts from Aristolochia baetica and Origanum compactumon human breast cancer cell line MCF-7. Pharm Biol. 2010; 48: 269-274, CrossRef.

Nevozhay D. Cheburator software for automatically calculating drug inhibitory concentrations from in vitro screening assays. PloS One. 2014; 9: 1-10, CrossRef.

Dwira S, Ariska TP, Fadilah F, Azizah NN, Erlina L. Comparison of cytotoxicity between ethyl acetate and ethanol extract of white turmeric (Kaempferia rotunda) rhizome extract against HeLa cervical cancer cell activity. Pharmacogn J. 2020; 12: 1297-302, CrossRef.

Prakash E, Gupta DK. Cytotoxic activity of ethanolic extract of Myristica fragrans (Houtt) against seven human cancer cell lines. Universal J Food Nutr Sci. 2013; 1: 1-3, CrossRef.

Thuong PT, Hung TM, Khoi NM, Nhung HTM, Chinh NT, Quy NT, et al. Cytotoxic and anti-tumor activities of lignans from the seeds of Vietnamese nutmeg Myristica fragrans. Arch Pharm Res. 2014; 37: 399-403, CrossRef.

Cuong TD, Lim CJ, Trang TTT, Bae YH, Thu NV, Tung NT, et al. Compounds from the seeds of Myristica fragrans and their cytotoxic activity. Nat Prod Sci. 2012; 18: 97-101, article.

Lee JW, Choi YH, Yoo MY, Choi SU, Hong KS, Lee BH, et al. Inhibitory effect of the seed extract of myristica semen on the proliferation of human tumor cell lines (II). Korean J Pharmacogn. 2006; 37: 206-11, article.

Chumkaew P, Srisawat T. New neolignans from the seeds of Myristica fragrans and their cytotoxic activities. J Nat Med. 2019; 73: 273-77, CrossRef.

Maheswari U, Ghosh K, Sadras SR. Licarin A induces cell death by activation of autophagy and apoptosis in non-small cell lung cancer cells. Apoptosis. 2018; 23: 210-25, CrossRef.

Min BS, Cuong TD, Hung TM, Min BK, Shin BS, Me HW. Inhibitory effect of lignans from myristica fragrans on LPS-induced NO production in RAW264.7 Cells. Bull Korean Chem Soc. 2011; 32: 4059-62, CrossRef.

Park S, Lee DK, Yang CH. Inhibition of fos–jun–DNA complex formation by dihydroguaiaretic acid and in vitro cytotoxic effects on cancer cells. Cancer Lett. 1998; 127: 23–28, CrossRef.

Sharifi-Rad J, Sureda A, Tenore GC, Daglia M, Sharifi-Rad M, Valussi M, et al. Biological activities of essential oils: from plant chemoecology to traditional healing systems. Molecules. 2017; 22: 1-55, CrossRef.

Lu X, Yang L, Chen J, et al. The action and mechanism of myrislignan on A549 cells in vitro and in vivo. J Nat Med. 2017; 71: 76-85, CrossRef.

Wicaksono BD, Tangkearung E, Sandra F. Brucea javanica leaf extract induced apoptosis in human oral squamous cell carcinoma (HSC2) cells by attenuation of mitochondrial membrane permeability. Indones Biomed J. 2015; 7: 107-10, CrossRef.

Rizal MI, Sandra F. Brucea javanica leaf extract activates caspase-9 and caspase-3 of mitochondrial apoptotic pathway in human oral squamous cell carcinoma. Indones Biomed J. 2016; 8: 43-8, CrossRef.

Yuan J. Caspases in apoptosis and beyond. Oncogene. 2008; 27: 6194-206, CrossRef.


Indexed by:






The Prodia Education and Research Institute