Sains Malaysiana 52(9)(2023): 2657-2671

http://doi.org/10.17576/jsm-2023-5209-15

Dioscorea alata Extract as Anti-Cancer Agent by Regulating Genetic Mutations, Apoptosis, and Cell Proliferation: in vitro and in silico Studies

(Ekstrak Dioscorea alata sebagai Agen Anti-Kanser dengan Mengawal Mutasi Genetik, Apoptosis dan Pembiakan Sel: Kajian in vitro dan in silico)

SRI NABAWIYATI NURUL MAKIYAH^1,*, YUYUN IKA CHRISTINA² & AHMAD HAFIDUL AHKAM²

¹Department of Histology and Immunology, School of Medicine, Faculty of Medicine and Health Sciences, Universitas Muhammadiyah Yogyakarta, Yogyakarta 55183, Indonesia

²Biology Department, Faculty of Mathematics and Life Sciences, Brawijaya University, Indonesia

Diserahkan: 23 Disember 2022/Diterima: 5 September 2023

Abstract

Dioscorea alata (DA) tubers contain some compounds of potential anti-cancer agents. However, despite its potential, there has been limited investigation on the potential anti-cancer agent of DA, especially in breast cancer cells. Therefore, this study aims to investigate the anti-cancer effect of ethanolic extract of DA tubers against the MCF-7 cell lines in vitro and in silico. The in vitro study was carried out by cytotoxicity effect using MTT assay. The in silico study examined active ingredients, determined anti-cancer activity, analyzed the protein target, protein-protein interactions, and molecular docking. The study results showed that the high-concentration ethanolic extract of DA tubers exhibited the highest decrease in MCF-7 cell viability with an IC₅₀ value of 50.98 g/mL and can be categorized as a promising anti-cancer agent. As many as 34 active compounds belonging to anthocyanins, saponins, and flavonoids were screened. Active compounds of DA tubers with a violation score < 2 were analyzed for their biological potentials. Ten compounds of DA with the highest potency were selected for docking analysis with the targeted protein EGFR. The in-silico study showed that 21 bioactive compounds contribute to the anti-cancer activity of DA. Chlorogenic acid is the most promising bioactive compound as an anti-cancer since it is the most similar to the anti-cancer drug. Meanwhile, dihydroquercetin has a low binding affinity value, requiring less energy to bind to proteins in cancer signaling indicated that this compound is promising as anti-cancer. This study suggested that the ethanolic extract of DA tubers can be considered an anti-cancer agent against the MCF-7 cell line in vitro by regulating genetic mutations, apoptosis, and cell proliferation.

Keywords: Anti-cancer; chlorogenic acid; dihydroquercetin; Dioscorea alata; in silico

Abstrak

Ubian Dioscorea alata (DA) mengandungi beberapa sebatian agen anti-kanser yang berpotensi. Walaupun dengan potensinya, terdapat penyelidikan yang terhad mengenai potensi agen anti-kanser DA, terutamanya dalam sel kanser payudara. Oleh itu, kajian ini bertujuan untuk mengkaji kesan anti-kanser ekstrak etanol ubian DA terhadap garisan sel MCF-7 secara in vitro dan in silico. Kajian in vitro telah dijalankan melalui kesan ketoksikan menggunakan asai MTT. Kajian in silico mengkaji bahan aktif menentukan aktiviti anti-kanser, menganalisis sasaran protein, interaksi protein dan dok molekul. Keputusan kajian menunjukkan bahawa ekstrak etanol berkepekatan tinggi tuber DA mengeluarkan penurunan tertinggi dalam daya maju sel MCF-7 dengan nilai IC₅₀ sebanyak 50.98 g/mL dan boleh dikategorikan sebagai agen anti-kanser yang berpotensi. Sebanyak 34 sebatian aktif kepunyaan antosianin, saponin dan flavonoid telah disaring. Sebatian aktif ubian DA dengan skor pelanggaran < 2 telah dianalisis untuk potensi biologinya. Sepuluh sebatian DA dengan potensi tertinggi telah dipilih untuk analisis dok dengan protein yang disasarkan EGFR. Kajian in silico menunjukkan bahawa 21 sebatian bioaktif menyumbang kepada aktiviti anti-kanser DA. Asid klorogenik adalah sebatian bioaktif yang paling berpotensi sebagai anti-kanser kerana ia adalah yang paling sama dengan ubat anti-kanser. Sementara itu, dihidrokuersetin mempunyai nilai pertalian pengikatan yang rendah, memerlukan kurang tenaga untuk mengikat protein dalam isyarat kanser menunjukkan bahawa sebatian ini berpotensi sebagai anti-kanser. Penyelidikan ini mencadangkan bahawa ekstrak etanol ubian DA boleh dianggap sebagai agen anti-kanser terhadap garisan sel MCF-7 secara in vitro dengan mengawal mutasi genetik, apoptosis dan percambahan sel.

Kata kunci: Anti-kanser; asid klorogenik; dihidrokuersetin; Dioscorea alata; in silico

RUJUKAN

Amir, H. & Murcitro, B.G. 2017. Uji Microtetrazolium (MTT) ekstrak metanol daun Phaleria macrocarpa (Scheff.) Boerl terhadap sel kanker payudara MCF-7. Jurnal Pendidikan dan Ilmu Kimia 1(1): 27-32.

Aumsuwan, P., Khan, S.I., Khan, I.A., Ali, Z., Avula, B., Walker, L.A., Shariat-Madar, Z., Helferich, W.G., Katzenellenbogen, B.S. & Dasmahapatra, A.K. 2016. The anticancer potential of steroidal saponin, dioscin, isolated from wild yam (Dioscorea villosa) root extract in invasive human breast cancer cell line MDA-MB-231 in vitro. Archives of Biochemistry and Biophysics 591: 98-110. https://doi.org/10.1016/J.ABB.2015.12.001

Benet, L.Z., Hosey, C.M., Ursu, O. & Oprea, T.I. 2016. BDDCS, the Rule of 5 and drugability. Advanced Drug Delivery Reviews 101: 89-98. https://doi.org/10.1016/J.ADDR.2016.05.007

Carocho, M. & Ferreira, I. 2013. The role of phenolic compounds in the fight against cancer-a review. Anti-Cancer Agents in Medicinal Chemistry 13(8): 1236-1258. https://doi.org/10.2174/18715206113139990301

Chen, D., Zheng, X., Kang, D., Yan, B., Liu, X., Gao, Y. & Zhang, K. 2012. Apoptosis and expression of the Bcl-2 family of proteins and P53 in human pancreatic ductal adenocarcinoma. Medical Principles and Practice: International Journal of the Kuwait University, Health Science Centre 21(1): 68-73. https://doi.org/10.1159/000332423

Dallakyan, S. & Olson, A.J. 2015. Small-molecule library screening by docking with PyRx. In Chemical Biology. Methods in Molecular Biology, Vol. 1263, edited by Hempel, J., Williams, C. & Hong, C. New York: Humana Press. pp. 243-250. https://doi.org/10.1007/978-1-4939-2269-7_19

Dey, P., Roy Chowdhuri, S., Sarkar, M.P. & Chaudhuri, T.K. 2016. Evaluation of anti-inflammatory activity and standardisation of hydro-methanol extract of underground tuber of Dioscorea alata. Pharmaceutical Biology 54(8): 1474-1482. https://doi.org/10.3109/13880209.2015.1104702

Filimonov, D.A., Lagunin, A.A., Gloriozova, T.A., Rudik, A.V., Druzhilovskii, D.S., Pogodin, P.V. & Poroikov, V.V. 2014. Prediction of the biological activity spectra of organic compounds using the pass online web resource. Chemistry of Heterocyclic Compounds 50(3): 444-457. https://doi.org/10.1007/S10593-014-1496-1

Ghose, A.K., Viswanadhan, V.N. & Wendoloski, J.J. 1999. A knowledge-based approach in designing combinatorial or medicinal chemistry libraries for drug discovery. 1. A qualitative and quantitative characterization of known drug databases. Journal of Combinatorial Chemistry 1(1): 55-68. https://doi.org/10.1021/CC9800071/ASSET/IMAGES/MEDIUM/CC9800071N00001.GIF

Ha, T.K., Jung, I., Kim, M.E., Bae, S.K. & Lee, J.S. 2017. Anti-cancer activity of myricetin against human papillary thyroid cancer cells involves mitochondrial dysfunction-mediated apoptosis. Biomedicine & Pharmacotherapy 91: 378-384. https://doi.org/10.1016/j.biopha.2017.04.100

Hanahan, D. & Weinberg, R.A. 2000. The hallmarks of cancer. Cell 100(1): 57-70. https://doi.org/10.1016/S0092-8674(00)81683-9

Hazafa, A., Rehman, K.U., Jahan, N. & Jabeen, Z. 2020. The role of polyphenol (flavonoids) compounds in the treatment of cancer cells. Nutrition and Cancer 72(3): 386-397. https://doi.org/10.1080/01635581.2019.1637006

Keiser, M.J., Roth, B.L., Armbruster, B.N., Ernsberger, P., Irwin, J.J. & Shoichet, B.K. 2007. Relating protein pharmacology by ligand chemistry. Nature Biotechnology 25(2): 197-206. https://doi.org/10.1038/nbt1284

Kemenkes Republik Indonesia. 2022. Kanker Payudara Paling Banyak di Indonesia, Kemenkes Targetkan Pemerataan Layanan Kesehatan. Https://Www.Kemkes.Go.Id/Article/View/22020400002/Kanker-Payudara-Paling-Banyak-Di-Indonesia-Kemenkes-Targetkan-Pemerataan-Layanan-Kesehatan.Html

Ko, E.Y. & Moon, A. 2015. Natural products for chemoprevention of breast cancer. Journal of Cancer Prevention 20(4): 223. https://doi.org/10.15430/JCP.2015.20.4.223

Li, W., Peng, C., Zhaojie, L. & Wei, W. 2022. Chemopreventive and therapeutic properties of anthocyanins in breast cancer: A comprehensive review. Nutrition Research 107: 48-64. https://doi.org/10.1016/j.nutres.2022.08.005

Lipinski, C.A., Lombardo, F., Dominy, B.W. & Feeney, P.J. 2001. Experimental and computational approaches to estimate solubility and permeability in drug discovery and development settings. Advanced Drug Delivery Reviews 46(1-3): 3-26. https://doi.org/10.1016/S0169-409X(00)00129-0

Łukasiewicz S, Czeczelewski M, Forma A, Baj J, Sitarz R and Stanisławek A. Breast Cancer-Epidemiology, Risk Factors, Classification, Prognostic Markers, and Current Treatment Strategies-An Updated Review. Cancers (Basel). 2021 Sep; 13(17): 4287. doi: 10.3390/cancers13174287

Makiyah, S.N.N., Kita, M., Setyawati, I. & Tasminatun, S. 2022. Dioscorea alata L. tubers improve diabetes through anti-hyperglycemia, anti-inflammation, ameliorate insulin resistance and mitochondrial dysfunction. The Indonesian Biomedical Journal 14(4): 365-375. https://doi.org/10.18585/inabj.v14i4.1966

Mardiyaningsih & Ismiyati. 2014. Aktivitas sitotoksik ekstrak etanolik daun alpukat (Persea americana Mil.) pada sel kanker leher rahim HeLa. Tradisional Medicine Journal 19(1): 141-150.

Mirulaini, S. & Shahira. 2011. Novel effect of diosgenin-A plant derived steroid: A review. Pharmacology Online 1: 726-736.

Molinspiration Cheminformatics. 2010. Molinspiration Cheminformatics.

Naeem A, Hu P, Yang M, Zhang J, Liu Y, Zhu W, Zheng Q. Natural Products as Anticancer Agents: Current Status and Future Perspectives. Molecules. 2022; 27(23):8367. https://doi.org/10.3390/molecules27238367

Negrini, S., Gorgoulis, V.G. & Halazonetis, T.D. 2010. Genomic instability - An evolving hallmark of cancer. Nature Reviews Molecular Cell Biology 11(3): 220-228. https://doi.org/10.1038/nrm2858

Olayemi, J.O. & Ajaiyeoba, E.O. 2010. Anti-inflammatory studies of yam (Dioscorea esculenta) extract on Wistar rats. African Journal of Biotechnology 6(16): 1913-1915. https://doi.org/10.4314/ajb.v6i16.57831

Pearce, A., Haas, M., Viney, R., Pearson, S.A., Haywood, P., Brown, C. & Ward, R. 2017. Incidence and severity of self-reported chemotherapy side effects in routine care: A prospective cohort study. PLoS ONE 12(10): e0184360. https://doi.org/10.1371/JOURNAL.PONE.0184360

Petricca, S., Celenza, G., Costagliola, C., Tranfa, F. & Iorio, R. 2022. Cytotoxicity, mitochondrial functionality, and redox status of human conjunctival cells after short and chronic exposure to preservative-free bimatoprost 0.03% and 0.01%: An in vitro comparative study. International Journal of Molecular Sciences 23(22): 14113. https://doi.org/10.3390/ijms232214113

Pollastri, M.P. 2010. Overview on the rule of five. Current Protocols in Pharmacology 49(1): 9.12.1-9.12.8. https://doi.org/10.1002/0471141755.PH0912S49

Raju, J. & Rao, C.V. 2012. Diosgenin, a steroid saponin constituent of yams and fenugreek: Emerging evidence for applications in medicine. Bioactive Compounds in Phytomedicine. InTech. https://doi.org/10.5772/26700

Shannon, P., Markiel, A., Ozier, O., Baliga, N.S., Wang, J.T., Ramage, D., Amin, N., Schwikowski, B. & Ideker, T. 2003. Cytoscape: A software environment for integrated models of biomolecular interaction networks. Genome Research 13(11): 2498-2504. https://doi.org/10.1101/gr.1239303

Sharma, T., Singh, D., Mahapatra, A., Mohapatra, P., Sahoo, S. & Sahoo, S.K. 2022. Advancements in clinical translation of flavonoid nanoparticles for cancer treatment. OpenNano 8: 100074. https://doi.org/10.1016/j.onano.2022.100074

Smolarz B, Nowak AZ, Romanowicz H. Breast Cancer-Epidemiology, Classification, Pathogenesis and Treatment (Review of Literature). Cancers (Basel). 2022 May 23;14(10):2569. doi: 10.3390/cancers14102569.

Stasevych, M., Viktor, Z. & Novikov, V. 2020. A computational approach in the search of new biologically active 9,10-anthraquinone derivatives. 3rd International Conference on Informatics & Data-Driven Medicine, November 19-21. Växjö, Sweden.

Sun, F., Zheng, X.Y., Ye, J., Wu, T.T., Wang, J.I. & Chen, W. 2012. Potential anticancer activity of myricetin in human T24 bladder cancer cells both in vitro and in vivo. Nutrition and Cancer 64(4): 599-606. https://doi.org/10.1080/01635581.2012.665564

Sung, H., Ferlay, J., Siegel, R.L., Laversanne, M., Soerjomataram, I., Jemal, A. & Bray, F. 2021. Global Cancer Statistics 2020: GLOBOCAN estimates of incidence and mortality worldwide for 36 cancers in 185 countries. CA: A Cancer Journal for Clinicians 71(3): 209-249. https://doi.org/10.3322/caac.21660

Supriatno, Nurlelasari, Herlina, T., Harneti, D., Maharani, R., Hidayat, A.T., Mayanti, T., Supratman, U., Azmi, M.N. & Shiono, Y. 2018. A new limonoid from stem bark of Chisocheton pentandrus (Meliaceae). Natural Product Research 32(21): 2610-2616. https://doi.org/10.1080/14786419.2018.1428600

Ueda, J.Y., Tezuka, Y., Banskota, A.H., le Tran, Q., Tran, Q.K., Harimaya, Y., Saiki, I. & Kadota, S. 2002. Antiproliferative activity of Vietnamese medicinal plants. Biological & Pharmaceutical Bulletin 25(6): 753-760. https://doi.org/10.1248/BPB.25.753

Ullah, A., Munir, S., Badshah, S.L., Khan, N., Ghani, L., Poulson, B.G., Emwas, A-H. & Jaremko, M. 2020. Important flavonoids and their role as a therapeutic agent. Molecules 25(22): 5243. https://doi.org/10.3390/molecules25225243

Virgili, F., Acconcia, F., Ambra, R., Rinna, A., Totta, P. & Marino, M. 2004. Nutritional flavonoids modulate estrogen receptor alpha signaling. IUBMB Life 56(3): 145-151. https://doi.org/10.1080/15216540410001685083

Wallace, K., Asemota, H. & Gray, W. 2021. Acetone extract of Dioscorea alata inhibits cell proliferation in cancer cells. American Journal of Plant Sciences 12(03): 300-314. https://doi.org/10.4236/ajps.2021.123019

Wee, P. & Wang, Z. 2017. Epidermal growth factor receptor cell proliferation signaling pathways. Cancers 9(5): 52. https://doi.org/10.3390/cancers9050052

*Pengarang untuk surat-menyurat; email: nurul.makiyah@umy.ac.id