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BMC Complementary Medicine and Therapies
Published in: BMC Complementary Medicine and Therapies 1/2020

01-12-2020 | Prostate Cancer | Research article

Selective cytotoxic and anti-metastatic activity in DU-145 prostate cancer cells induced by Annona muricata L. bark extract and phytochemical, annonacin

Authors: Kimberley Foster, Omolola Oyenihi, Sunelle Rademan, Joseph Erhabor, Motlalepula Matsabisa, James Barker, Moses K. Langat, Amy Kendal-Smith, Helen Asemota, Rupika Delgoda

Published in: BMC Complementary Medicine and Therapies | Issue 1/2020

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Abstract

Background

Annona muricata L. was identified as a popular medicinal plant in treatment regimens among cancer patients in Jamaica by a previously conducted structured questionnaire. Ethnomedically used plant parts, were examined in this study against human prostate cancer cells for the first time and mechanisms of action elucidated for the most potent of them, along with the active phytochemical, annonacin.

Methods

Nine extracts of varying polarity from the leaves and bark of A. muricata were assessed initially for cytotoxicity using the MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) assay on PC-3 prostate cancer cells and the ethyl acetate bark (EAB) extract was identified as the most potent. EAB extract was then standardized for annonacin content using High-performance Liquid Chromatography - Mass Spectrometry (HPLC-MS) and shown to be effective against a second prostate cancer cell line (DU-145) also. The mode of cell death in DU-145 cells were assessed via several apoptotic assays including induction of increased reactive oxygen species (ROS) production, reduction of mitochondrial membrane potential, activation of caspases and annexin V externalization combined with morphological observations using confocal microscopy. In addition, the potential to prevent metastasis was examined via inhibition of cell migration, vascular endothelial growth factor (VEGF) and angiogenesis using the chorioallantoic membrane assay (CAM).

Results

Annonacin and EAB extract displayed selective and potent cytotoxicity against the DU-145 prostate carcinoma cells with IC50 values of 0.1 ± 0.07 μM and 55.501 ± 0.55 μg/mL respectively, without impacting RWPE-1 normal prostate cells, in stark contrast to chemotherapeutic docetaxel which lacked such selectivity. Docetaxel’s impact on the cancerous DU-145 was improved by 50% when used in combination with EAB extract. Insignificant levels of intracellular ROS content, depolarization of mitochondrial membrane, Caspase 3/7 activation, annexin V content, along with stained morphological evaluations, pointed to a non-apoptotic mode of cell death. The extract at 50 μg/mL deterred cell migration in the wound-healing assay, while inhibition of angiogenesis was displayed in the CAM and VEGF inhibition assays for both EAB (100 μg /mL) and annonacin (0.5 μM).

Conclusions

Taken together, the standardized EAB extract and annonacin appear to induce selective and potent cell death via a necrotic pathway in DU-145 cells, while also preventing cell migration and angiogenesis, which warrant further examinations for mechanistic insights and validity in-vivo.
Literature
1.
Delgoda R, Murray JE. Evolutionary perspective on the role of plant secondary metabolites In: Badal S and Delgoda R, Editors. Pharmacognosy: fundamentals, applications and strategies. London: Elsevier Academic Press; 2016. p. 93–100.
2.
Organization WH. WHO traditional medicine strategy 2002–2005. Geneva: World Health Organization; 2002.
3.
Picking D, Younger N, Mitchell S, Delgoda R. The prevalence of herbal medicine home use and concomitant use with pharmaceutical medicines in Jamaica. J Ethnopharmacol. 2011;137(1):305–11.PubMedCrossRef
4.
Newman DJ, Cragg GM. Natural products as sources of new drugs over the nearly four decades from 01/1981 to 09/2019. J Nat Prod. 2020;83(3):770–803.
5.
Rady I, Bloch MB, Chamcheu R-CN, Banang Mbeumi S, Anwar MR, Mohamed H, Babatunde AS, Kuiate J-R, Noubissi FK, El Sayed KA. Anticancer properties of Graviola (Annona muricata): a comprehensive mechanistic review. Oxidative Med Cell Longev. 2018;2018:1–39. https://​doi.​org/​10.​1155/​2018/​1826170.
6.
Cragg, GM, Kingston DGI, Newman DJ, eds. Anticancer agents from natural products. Boca Raton: CRC press/Taylor & Francis Group; 2011.
7.
Picking D, Delgoda R, Vandebroek I. Traditional knowledge systems and the role of traditional medicine in Jamaica. CAB Rev. 2019;14(045):1–13.CrossRef
8.
Williams LA, Rosner H, Kraus W. Molecules with potential for cancer therapy in the developing world: Dibenzyl trisulfide (DTS). In: Genomics Applications for the Developing World. New York, NY; Springer; 2012. p. 273–8.
9.
Lowe H, Toyang N, Bryant J. In vitro and in vivo anti-cancer effects of Tillandsia recurvata (ball Moss) from Jamaica. West Indian Med J. 2013;62(3):177–80.PubMed
10.
Campbell S, Murray J, Delgoda R, Gallimore W. Two new Oxodolastane Diterpenes from the Jamaican macroalga Canistrocarpus cervicornis. Mar Drugs. 2017;15(6):150.PubMedCentralCrossRef
11.
Foster K, Younger N, Aiken W, Brady-West D, Delgoda R. Reliance on medicinal plant therapy among cancer patients in Jamaica. Cancer Causes Control. 2017;28(11):1349–56.PubMedCrossRef
12.
Fidianingsih I, Handayani ES. Annona muricata aqueous extract suppresses T47D breast cancer cell proliferation. Univ Med. 2015;33(1):19–26.
13.
Clement YN, Mahase V, Jagroop A, Kissoon K, Maharaj A, Mathura P, Mc Quan C, Ramadhin D, Mohammed C. Herbal remedies and functional foods used by cancer patients attending specialty oncology clinics in Trinidad. BMC Complement Altern Med. 2016;16(1):399.PubMedPubMedCentralCrossRef
14.
Jacobo-Herrera NJ, Perez-Plasencia C, Castro-Torres VA, Martinez-Vazquez M, Gonzalez-Esquinca AR, Zentella-Dehesa A. The potential of acetogenins in cancer treatment. Front Pharmacol. 2019;10:783.PubMedPubMedCentralCrossRef
15.
Coria-Téllez AV, Montalvo-Gónzalez E, Yahia EM, Obledo-Vázquez EN. Annona muricata: a comprehensive review on its traditional medicinal uses, phytochemicals, pharmacological activities, mechanisms of action and toxicity. Arab J Chem. 2018;11(5):662–91.CrossRef
16.
17.
Alonso-Castro AJ, Villarreal ML, Salazar-Olivo LA, Gomez-Sanchez M, Dominguez F, Garcia-Carranca A. Mexican medicinal plants used for cancer treatment: pharmacological, phytochemical and ethnobotanical studies. J Ethnopharmacol. 2011;133(3):945–72.PubMedCrossRef
18.
Monigatti M, Bussmann RW, Weckerle CS. Medicinal plant use in two Andean communities located at different altitudes in the Bolívar Province, Peru. J Ethnopharmacol. 2013;145(2):450–64.PubMedCrossRef
19.
Yang C, Gundala SR, Mukkavilli R, Vangala S, Reid MD, Aneja R. Synergistic interactions among flavonoids and acetogenins in Graviola (Annona muricata) leaves confer protection against prostate cancer. Carcinogenesis. 2015;36(6):656–65.PubMedPubMedCentralCrossRef
20.
Fang XP, Rieser MJ, Gu ZM, Zhao GX, McLaughlin JL. Annonaceous acetogenins: an updated review. Phytochem Anal. 1993;4(1):27–48.CrossRef
21.
Al-Dabbagh B, Elhaty IA, Al Sakkaf R, El-Awady R, Ashraf SS, Amin A. Antioxidant and anticancer activities of Trigonella foenum-graecum, Cassia acutifolia and Rhazya stricta. BMC Complement Altern Med. 2018;18(1):1–12.CrossRef
22.
Lannuzel A, Michel P, Höglinger G, Champy P, Jousset A, Medja F, Lombes A, Darios F, Gleye C, Laurens A. The mitochondrial complex I inhibitor annonacin is toxic to mesencephalic dopaminergic neurons by impairment of energy metabolism. Neuroscience. 2003;121(2):287–96.PubMedCrossRef
23.
Yap CV, Subramaniam KS, Khor SW, Chung I. Annonacin exerts antitumor activity through induction of apoptosis and extracellular signal-regulated kinase inhibition. Pharm Res. 2017;9(4):378.
24.
Yiallouris A, Patrikios I, Johnson EO, Sereti E, Dimas K, De Ford C, Fedosova NU, Graier WF, Sokratous K, Kyriakou K. Annonacin promotes selective cancer cell death via NKA-dependent and SERCA-dependent pathways. Cell Death Dis. 2018;9(7):764.PubMedPubMedCentralCrossRef
25.
Roduan MRM, Hamid RA, Mohtarrudin N. Modulation of cancer signalling pathway (s) in two-stage mouse skin tumorigenesis by annonacin. BMC Complement Altern Med. 2019;19(1):238.CrossRef
26.
Raybaudi-Massilia R, Suárez AI, Arvelo F, Sojo F, Mosqueda-Melgar J, Zambrano A, Calderón-Gabaldón MI. An analysis in-vitro of the cytotoxic, antioxidant and antimicrobial activity of aqueous and alcoholic extracts of Annona muricata L. seed and pulp. Br J Appl Sci Technol. 2015;5(4):333.CrossRef
27.
Sulistyoningrum E, Rachmani EPN, Baroroh HN, Rujito L. Annona muricata leaves extract reduce proliferative indexes and improve histological changes in rat's breast cancer. J Appl Pharm Sci. 2017;7(01):149–55.CrossRef
28.
Moghadamtousi SZ, Karimian H, Rouhollahi E, Paydar M, Fadaeinasab M, Kadir HA. Annona muricata leaves induce G1 cell cycle arrest and apoptosis through mitochondria-mediated pathway in human HCT-116 and HT-29 colon cancer cells. J Ethnopharmacol. 2014;156:277–89.CrossRef
29.
Moghadamtousi SZ, Rouhollahi E, Karimian H, Fadaeinasab M, Firoozinia M, Abdulla MA, Kadir HA. The chemopotential effect of Annona muricata leaves against azoxymethane-induced colonic aberrant crypt foci in rats and the apoptotic effect of acetogenin annomuricin E in HT-29 cells: a bioassay-guided approach. PLoS One. 2015;10(4):e0122288.CrossRef
30.
Yuan S-SF, Chang H-L, Chen H-W, Yeh Y-T, Kao Y-H, Lin K-H, Wu Y-C, Su J-H. Annonacin, a mono-tetrahydrofuran acetogenin, arrests cancer cells at the G1 phase and causes cytotoxicity in a Bax-and caspase-3-related pathway. Life Sci. 2003;72(25):2853–61.PubMedCrossRef
31.
Supino R. MTT assays. In: O’Hare S, Atterwill CK. (eds) In Vitro Toxicity Testing Protocols. Methods in Molecular Biology™, vol 43. Humana Press; 1995. p.137–49.
32.
Van Meerloo J, Kaspers GJ, Cloos J. Cell sensitivity assays: the MTT assay. In: Cancer cell culture, Humana Press; 2011. p. 237–45.
33.
Koch H, Woodward J, Langat MK, Brown MJ, Stevenson PC. Flagellum removal by a nectar metabolite inhibits infectivity of a bumblebee parasite. Curr Biol. 2019;29(20):3494–500 e3495.PubMedCrossRef
34.
Simon H-U, Haj-Yehia A, Levi-Schaffer F. Role of reactive oxygen species (ROS) in apoptosis induction. Apoptosis. 2000;5(5):415–8.PubMedCrossRef
35.
Pieme CA, Kumar SG, Dongmo MS, Moukette BM, Boyoum FF, Ngogang JY, Saxena AK. Antiproliferative activity and induction of apoptosis by Annona muricata (Annonaceae) extract on human cancer cells. BMC Complement Altern Med. 2014;14(1):516.PubMedPubMedCentralCrossRef
36.
Bock FJ, Tait SW. Mitochondria as multifaceted regulators of cell death. Nat Rev Mol Cell Biol. 2020;21(2):85–100.
37.
Sirenko O, Hesley J, Rusyn I, Cromwell EF. High-content assays for hepatotoxicity using induced pluripotent stem cell–derived cells. Assay Drug Dev Technol. 2014;12(1):43–54.PubMedPubMedCentralCrossRef
38.
Lu C, Hassan H. Human stem cell factor-antibody [anti-SCF] enhances chemotherapy cytotoxicity in human CD34+ resistant myeloid leukaemia cells. Leuk Res. 2006;30(3):296–302.PubMedCrossRef
39.
Huang T-C, Lee J-F, Chen J-Y. Pardaxin, an antimicrobial peptide, triggers caspase-dependent and ROS-mediated apoptosis in HT-1080 cells. Marine drugs. 2011;9(10):1995–2009.PubMedPubMedCentralCrossRef
40.
Mahajan SD, Tutino VM, Redae Y, Meng H, Siddiqui A, Woodruff TM, Jarvis JN, Hennon T, Schwartz S, Quigg RJ. C5a induces caspase-dependent apoptosis in brain vascular endothelial cells in experimental lupus. Immunology. 2016;148(4):407–19.PubMedPubMedCentralCrossRef
41.
Liu K, Liu P-C, Liu R, Wu X. Dual AO/EB staining to detect apoptosis in osteosarcoma cells compared with flow cytometry. Med Sci Monit Basic Res. 2015;21:15.PubMedPubMedCentralCrossRef
42.
Wang H, Liu W, Black S, Turner O, Daniel JM, Dean-Colomb W, He QP, Davis M, Yates C. Kaiso, a transcriptional repressor, promotes cell migration and invasion of prostate cancer cells through regulation of miR-31 expression. Oncotarget. 2016;7(5):5677.PubMedCrossRef
43.
Bala A, Mukherjee P, Braga F, Matsabisa M. Comparative inhibition of MCF-7 breast cancer cell growth, invasion and angiogenesis by Cannabis sativa L. sourced from sixteen different geographic locations. S Afr J Bot. 2018;119:154–62.CrossRef
44.
Yang Z, Lu W, Ma X, Song D. Bioassay-guided isolation of an alkaloid with antiangiogenic and antitumor activities from the extract of Fissistigma cavaleriei root. Phytomedicine. 2012;19(3–4):301–5.PubMedCrossRef
45.
Guo J, Zhu T, Chen L, Nishioka T, Tsuji T, Xiao Z-XJ, Chen CY. Differential sensitization of different prostate cancer cells to apoptosis. Genes Cancer. 2010;1(8):836–46.PubMedPubMedCentralCrossRef
46.
Jayakumar S, Kunwar A, Sandur SK, Pandey BN, Chaubey RC. Differential response of DU145 and PC3 prostate cancer cells to ionizing radiation: role of reactive oxygen species, GSH and Nrf2 in radiosensitivity. Biochim Biophys Acta. 2014;1840(1):485–94.PubMedCrossRef
47.
Karanika S, Karantanos T, Kurosaka S, Wang J, Hirayama T, Yang G, Park S, Golstov AA, Tanimoto R, Li L. GLIPR1-ΔTM synergizes with docetaxel in cell death and suppresses resistance to docetaxel in prostate cancer cells. Mol Cancer. 2015;14(1):122.PubMedPubMedCentralCrossRef
48.
49.
50.
Alfano ACC, Paiva CE, Rugno FC, da Silva RH, Paiva BSR. Biologically based therapies are commonly self-prescribed by Brazilian women for the treatment of advanced breast cancer or its symptoms. Support Care Cancer. 2014;22(5):1303–11.PubMedCrossRef
51.
Almog L, Lev E, Schiff E, Linn S, Ben-Arye E. Bridging cross-cultural gaps: monitoring herbal use during chemotherapy in patients referred to integrative medicine consultation in Israel. Support Care Cancer. 2014;22(10):2793–804.PubMedCrossRef
52.
Alsanad SM, Howard RL, Williamson EM. An assessment of the impact of herb-drug combinations used by cancer patients. BMC Complement Altern Med. 2016;16(1):393.PubMedPubMedCentralCrossRef
53.
Green DR, Kroemer G. The pathophysiology of mitochondrial cell death. Science. 2004;305(5684):626–9.PubMedCrossRef
54.
Redza-Dutordoir M, Averill-Bates DA. Activation of apoptosis signalling pathways by reactive oxygen species. Biochim Biophys Acta. 2016;1863(12):2977–92.PubMedCrossRef
55.
Jäättelä M. Multiple cell death pathways as regulators of tumour initiation and progression. Oncogene. 2004;23(16):2746–56.PubMedCrossRef
56.
Laksmitawati DR, Prasanti AP, Larasinta N, Syauta GA, Hilda R, Ramadaniati HU, Widyastuti A, Karami N, Afni M, Rihibiha DD. Anti-inflammatory potential of gandarusa (Gendarussa vulgaris Nees) and soursoup (Annona muricata L) extracts in LPS stimulated-macrophage cell (RAW264. 7). J Nat Remedies. 2016;16(2):73–81.CrossRef
57.
Krysko O, De Ridder L, Cornelissen M. Phosphatidylserine exposure during early primary necrosis (oncosis) in JB6 cells as evidenced by immunogold labeling technique. Apoptosis. 2004;9(4):495–500.PubMedCrossRef
58.
59.
Torres MP, Rachagani S, Purohit V, Pandey P, Joshi S, Moore ED, Johansson SL, Singh PK, Ganti AK, Batra SK. Graviola: a novel promising natural-derived drug that inhibits tumorigenicity and metastasis of pancreatic cancer cells in vitro and in vivo through altering cell metabolism. Cancer Lett. 2012;323(1):29–40.PubMedPubMedCentralCrossRef
60.
Berghe TV, Grootjans S, Goossens V, Dondelinger Y, Krysko DV, Takahashi N, Vandenabeele P. Determination of apoptotic and necrotic cell death in vitro and in vivo. Methods. 2013;61(2):117–29.CrossRef
61.
Lamb HM. Double agents of cell death: novel emerging functions of apoptotic regulators. FEBS J. 2020;287(13):2647–63.
Metadata
Title
Selective cytotoxic and anti-metastatic activity in DU-145 prostate cancer cells induced by Annona muricata L. bark extract and phytochemical, annonacin
Authors
Kimberley Foster
Omolola Oyenihi
Sunelle Rademan
Joseph Erhabor
Motlalepula Matsabisa
James Barker
Moses K. Langat
Amy Kendal-Smith
Helen Asemota
Rupika Delgoda
Publication date
01-12-2020
Publisher
BioMed Central
Published in
BMC Complementary Medicine and Therapies / Issue 1/2020
Electronic ISSN: 2662-7671
DOI
https://doi.org/10.1186/s12906-020-03130-z

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