In vitro antitumor effect of essential oil of leaves from ñandypa (Genipa americana L.) on human pancreatic carcinoma cell lines (MIA PaCa2)
DOI:
https://doi.org/10.52379/mcs.v10.699Keywords:
phytoteraphy, Genipa americana L., MRC-5 cells, antitumoral, essential oilAbstract
Introduction: South American biodiversity offers a source of new therapeutic compounds. Cancer remains a major global health issue with high morbidity and mortality. ñandypa (Genipa americana L.), a medicinal plant with various reported properties, was tested for citotoxic activity of the essential oil from their leaves. Objective: to determine the in vitro antitumor effect of an essential oil derived from ñandypa on human pancreatic carcinoma cell lines (MIA PaCa2). Methodology:Essential oils from G. americana leaves were obtained via hydrodistillation, and their chemical composition was analyzed using Gas Chromatography-Flame Ionization Detection-Mass Spectrometry (GC-FID-MS). Pancreatic tumor cells and normal fibroblast cells were then tested for antitumor activity. The cells were treated with serial dilutions of the essential oil (0.1%, 0.5%, 1%, 1.5%, 5%, and 10%). Cell viability was measured using an MTT assay, with results read on a spectrophotometer at 570nm and 630nm. Results: GC-FID-MS analysis revealed the oil's primary components were alcohols (17.83%), aldehydes (9.58%), and esters (5.18%). Exposure to the oil for four hours resulted in a high rate of tumor cell mortality, with the effect being more pronounced at higher concentrations. Importantly, the oil was not found to be toxic to normal cells (MRC-5). Conclusion: The essential oil of Genipa americana L. exhibits significant, dose-dependent antitumor activity against human pancreatic carcinoma cells (MIA PaCa-2). Notably, it showed no toxicity toward normal human fibroblast cells (MRC-5). These findings, supported by the presence of bioactive compounds like BHT, ?-pinene, and ?-caryophyllene, suggest its potential as a selective therapeutic agent for pancreatic cancer.
Downloads
References
1. Maisonneuve P, Lowenfels A. Epidemiology and Prospects for Prevention of Pancreatic Cancer. Pancreatic Cancer. 2018;3–18. https://doi.org/10.1007/978-1-4939-7193-0_73
2. American Cancer Society. Key Statistics for Pancreatic Cancer. 2021. https://www.cancer.org/cancer/types/pancreatic-cancer/about/key-statistics.html
3. The Global Cancer Observatory. Pancreas fact sheet International Agency for Research on Cancer. 2020. https://gco.iarc.fr/today/data/factsheets/cancers/13-Pancreas-fact-sheet.pdf
4. Ribeiro Barbosa I, dos Santos CA, Bezerra de Souza DL. Pancreatic cancer in Brazil: mortality trends and projections until 2029. Arq Gastroenterol. 2018;55(3):230–6. https://doi.org/10.1590/S0004-2803.201800000-59
5. Wasmosy Monti JB. Cáncer de páncreas: Encare diagnóstico y terapéutico. Tendencias en Medicina. 2010. http://www.tendenciasenmedicina.com/Imagenes/imagenes05p/art_11.pdf
6. Dias RB, De Faro Valverde L, Rocha CAG, Bezerra DP. Principles of cancer pathogenesis and therapies: A brief overview. Bioactive Essential Oils and Cancer. 2015;1–17. https://doi.org/10.1007/978-3-319-19144-7_1
7. Grützmann R. Epidemiology, Treatment, and Outcome of Pancreatic Cancer. Molecular Diagnostics and Treatment of Pancreatic Cancer. 2014;3–9. https://doi.org/10.1016/B978-0-12-408103-1.00001-7
8. Rajawat J, Misra G. Overview of cancer assessment, diagnostics, and therapeutics. Protocol Handbook for Cancer Biology. 2021;1–6. https://doi.org/10.1016/B978-0-323-90006-5.00004-5
9. Burris III HA, Moore MJ, Cripps MC, Portenoy RK, Stephens CD, von Hoff DD. Improvements in Survival and Clinical Benefit With Gemcitabine as First Line Therapy for Patients With Advanced Pancreas Cancer: A Randomized Trial. Journal of Clinical Oncology. 1997;15(66):2403–13. https://doi.org/10.1200/JCO.1997.15.6.2403
10. Garrido-Laguna I, Hidalgo M. Pancreatic cancer: from state-of-the-art treatments to promising novel therapies. Nature Reviews Clinical Oncology. 2015;12(6):319–34. https://doi.org/10.1038/nrclinonc.2015.53
11. Sushma PS, Nagaraju GP. Pancreatic Cancer Resistance to Gemcitabine. Breaking Tolerance to Pancreatic Cancer Unresponsiveness to Chemotherapy. Indian J Med Res. 2019;45–56. https://doi.org/10.1016/B978-0-12-817661-0.00004-4
12. Missouri Botanical Garden. Genipa americana var. americana. Tropicos. 2021. https://www.tropicos.org/name/27906413
13. Keeler C. Genipa americana in native tropical medicine. Int J Dermatol. 1964;3(2):104–7. https://doi.org/10.1111/j.1365-4362.1964.tb06030.x
14. Nascimento LLL, de Oliveira Souza GG, da Silva Mendes JW, Calixto Donelardy AC, Viturino JJF, Gomes de Carvalho NK, et al. Genipa americana L.: A Review on Traditional Uses, Phytochemistry and Biological Activities. Chem Biodivers. 2024;21(11):e202400748. https://doi.org/10.1002/cbdv.202400748
15. Japan Internacional Cooperation Agency. Interim Report on Cooperation in Study (Chemical and Pharmaceutical Study on Herbs) with Paraguay. Asunción; 1987. https://openjicareport.jica.go.jp/pdf/10409878_01.pdf
16. Basualdo I, Soria N, Ortiz M, Degen R. Plantas medicinales comercializadas en los mercados de Asunción y Gran Asunción. Parte I. Rojasiana. 2004;6(1):95–114. https://www.qui.una.py/v2/wp-content/uploads/5_Plantas-medicinales-comercializadas.pdf
17. Pin A, González G, Marin G, Céspedes G, Cretton S, Christen P, et al. Plantas Medicinales del Jardín Botánico de Asunción. 1st ed. Asunción: Proyecto Etnobotanica Paraguaya; 2009
18. Fogel R, Céspedes C, López L, Valdez S, Soria N, Schmeda G. Propiedades medicinales de plantas: Conocimiento tradicional y patentes. Asunción;2016. https://www.conacyt.gov.py/sites/default/files/upload_editores/u294/Libro-INV-470-1.pdf
19. Aline Cigolini Ruzza D, Aparecida Bandini Rossi A, Martins Fernandes J, Cristina Moreno de Pedri E, Vicente Tiago A, Barboza Bispo R, et al. Etnobotânica do jenipapo (Genipa americana L., Rubiaceae) entre agricultores no município de Carlinda, Mato Grosso, Brasil. Brazilian Journal of Development. 2020;6(8):6118–84. https://doi.org/10.34117/bjdv6n8-509
20. Ruzza DAC, Rossi AAB, Fernandes JM, Pedri ECM de, Tiago AV, Bispo RB, Martins KC. Etnobotânica do jenipapo (Genipa americana L., Rubiaceae) entre agricultores no município de Carlinda, Mato Grosso, Brasil. Braz. J. Develop. 2020. 24;6(8):61161-84. https://ojs.brazilianjournals.com.br/ojs/index.php/BRJD/article/view/15462
21. Cáceres A MS, Machaín Singer M. Manual de uso de hierbas medicinales del Paraguay - UNESCO Digital Library. UNESCO; 2001. https://unesdoc.unesco.org/ark:/48223/pf0000156204
22. Brandão F, Santos D, Lima Ramos MI, Miyagusku L. Antimicrobial activity of hydroalcoholic extracts from genipap, baru and taruma. Ciência Rural. 2017;47(8):8. https://doi.org/10.1590/0103-8478cr20160252
23. Schneider GF, Salazar D, Hildreth SB, Helm RF, Whitehead SR. Comparative Metabolomics of Fruits and Leaves in a Hyperdiverse Lineage Suggests Fruits Are a Key Incubator of Phytochemical Diversification. Frontiers in Plant Science. 2021;12. https://www.frontiersin.org/journals/plant-science/articles/10.3389/fpls.2021.693739/full
24. Campos S, Shirley, Natan MB, de, Santana A, Dias E, et al. In vitro cytotoxicity and biological activities of Genipa americana (Rubiaceae) ethanolic extracts. African Journal of Microbiology Research. 2017;11(9):385–90. https://academicjournals.org/journal/AJMR/article-full-text-pdf/8E6E49263095
25. Sobolewska D, Galanty A, Grabowska K, Justyna Makowska-Wąs, Podolak I, Dagmara Wróbel-Biedrawa. Genipin—Simple but Significant Bioactive Iridoid for Therapeutical Application and Beyond: A Review. Life. 2025;15(2):159–9. https://doi.org/10.3390/life15020159
26. Ahmed R, Noor, Wang M, Iqbal S, Yi J, Yacine Hemar. Genipin, a natural blue colorant precursor: Source, extraction, properties, and applications. Food Chemistry. 2023;434:137498–8. https://doi.org/10.1016/j.foodchem.2023.137498
27. Claro RO, Rivero-Wendt CLG, Miranda-Vilela AL, Grisolia CK, Facco GG, Moreira D de L, et al. Toxicological effects of aqueous extract of Genipa americana L. leaves on adult zebrafish (Danio rerio): Chemical profile, histopathological effects and lack of genotoxicity. Toxicon. 2023;235(107305):107305. https://doi.org/10.1016/j.toxicon.2023.107305
28. Vasconcelos AL, Santos A v., Padilha RJR, Alves LC, Randau KP. Anatomical characterization of ultra-structures, biominerals and histolocalization of metabolites in leaves of Genipa americana. Revista Brasileira de Farmacognosia. 2017;27(5):541–8. https://doi.org/10.1016/j.bjp.2017.05.002
29. Alves JSF, de Medeiros LA, Fernandes-Pedrosa M de F, Araújo RM, Zucolotto SM. Iridoids from leaf extract of genipa americana. Brazilian Journal of Pharmacognosy. 2017;27(5):641–4. https://doi.org/10.1016/j.bjp.2017.03.006
30. Silva LMP, Alves JSF, da Silva Siqueira EM, de Souza Neto MA, Abreu LS, Tavares JF, et al. Isolation and Identification of the Five Novel Flavonoids from Genipa americana Leaves. Molecules: A Journal of Synthetic Chemistry and Natural Product Chemistry. 2018 Oct;23(10). https://doi.org/10.3390/molecules23102521
31. Lima JKA, Chicuta CPDL, Costa M de M, Costa MLA da, Grillo LAM, Santos AF dos, et al. Biotoxicity of aqueous extract of Genipa americana L. bark on red flour beetle Tribolium castaneum (Herbst). Ind Crops Prod. 2020;156:112874. https://doi.org/10.1016/j.indcrop.2020.112874
32. Nonato DTT, Aragão GF, Craveiro RMCB, Pereira MG, Vasconcelos SMM, Wong DVT, et al. Polysaccharide-rich extract of Genipa americana leaves protects seizures and oxidative stress in the mice model of pentylenetetrazole-induced epilepsy. Biomed Pharmacother. 2024;172(116212):116212. https://doi.org/10.1016/j.biopha.2024.116212
33. Rashid MU, Hussain I, Jehanzeb S, Ullah W, Ali S, Jain AG, et al. Pancreatic Cancer and Possible Therapeutic Options. Breaking Tolerance to Pancreatic Cancer Unresponsiveness to Chemotherapy. 2019 Jan;57–85. https://doi.org/10.1016/B978-0-12-817661-0.00005-6.
34. Guenther E, Ph D. The Essential Oils. Nature. 1949;163(4148):663–663. https://ia803001.us.archive.org/13/items/TheEssentialOilsVol1HisErnestGuenther/The%20Essential%20Oils%20-%20Vol%201_%20His%20-%20Ernest%20Guenther.pdf
35. Stahl E, Schild W. Pharmazeutische Biologie. Bd. 4, Drogenanalyse II, Inhaltsstoffe und Isolierungen. Stuttgart: Gustav Fischer Verlag; 1981. https://www.worldcat.org/es/title/pharmazeutische-biologie-bd-4-drogenanalyse-ii-inhaltsstoffe-und-isolierungen/oclc/222368498
36. Adams RP. Identification of essential oil components by gas chromatography/mass spectrometry. 4th ed. Illinois: Allured Publishing Corporation; 2009. https://www.cabdirect.org/cabdirect/abstract/20083116584
37. Aydin E, Türkez H, Geyikoğlu F. Antioxidative, anticancer and genotoxic properties of α-pinene on N2a neuroblastoma cells. Biologia. 2013;68(5):1004–9. https://doi.org/10.2478/s11756-013-0230-2
38. MRC-5 | ATCC. 2025. https://www.atcc.org/products/ccl-171
39. Jacobs JP, Jones CM, Baille JP. Characteristics of a Human Diploid Cell Designated MRC-5. Nature. 1970;227(5254):168–70. https://pubmed.ncbi.nlm.nih.gov/4316953/
40. Mosmann T. Rapid colorimetric assay for cellular growth and survival: Application to proliferation and cytotoxicity assays. Journal of Immunological Methods. 1983;65(1-2):55–63. https://pubmed.ncbi.nlm.nih.gov/6606682/
41. El-Abid H, Amaral C, Cunha SC, Augusto T v., Fernandes JO, Correia-da-Silva G, et al. Chemical composition and anti-cancer properties of Juniperus oxycedrus L. essential oils on estrogen receptor-positive breast cancer cells. J Funct Foods. 2019;59:261–71. https://doi.org/10.1016/j.jff.2019.05.042
42. Borges ES, Rezende CM. Main Aroma Constituents of Genipap (Genipa americana L.) and Bacuri (Platonia insignis M.). Journal of Essential Oil Research. 2011;12(1):71–4. https://doi.org/10.1080/10412905.2000.9712046
43. Miranda NC, Araujo ECB, Justino AB, Cariaco Y, Mota CM, Costa-Nascimento LA, et al. Anti-parasitic activity of Annona muricata L. leaf ethanolic extract and its fractions against Toxoplasma gondii in vitro and in vivo. Journal of Ethnopharmacology. 2021;273:114019. https://www.sciencedirect.com/science/article/abs/pii/S0378874121002464?via%3Dihub
44. Silva P, Codignoto C, Batista De Araújo S, Munhoz Bastos N, De T, Fernandes O, et al. In vitro cytotoxicity and biological activities of Genipa americana (Rubiaceae) ethanolic extracts. African Journal of Microbiology Research. 2017;11(9):385–90. https://doi.org/10.5897/AJMR2016.8418
45. Kuriakose GC, Arathi BP, Divya Lakshmanan M, Jiby M v., Gudde RS, Jayabhaskaran C. Sub-acute Toxicity Assessment of Taxol Isolated From Fusarium Solani, an Endophytic Fungus of Taxus Brevifolia, in Wistar Rats and Analyzing Its Cytotoxicity and Apoptotic Potential in Lung Cancer Cells. Front Oncol. 2020;10:1825. https://doi.org/10.3389/fonc.2020.538865
46. Amrutkar M, Gladhaug I. Pancreatic Cancer Chemoresistance to Gemcitabine. Cancers. 2017;9(12):157. https://pubmed.ncbi.nlm.nih.gov/29144412/
47. Zeng S, Pöttler M, Lan B, Grützmann R, Pilarsky C, Yang H. Chemoresistance in Pancreatic Cancer. International Journal of Molecular Sciences. 2019;20(18):4504. https://www.mdpi.com/1422-0067/20/18/4504/html
48. de Jesus AS, Coelho CR, Barreto IC, Filho JGS, Cesar P, Nogueira L, et al. Composition and Bioactivity of Essential Oil From the Leaves of Genipa americana Against the Coconut Mite Aceria guerreronis. Journal of Agricultural Science. 2019;11(18):197–205. https://doi.org/10.5539/jas.v11n18p197
49. Figueiredo AC, Barroso JG, Pedro LG, Scheffer JJC. Factors affecting secondary metabolite production in plants: volatile components and essential oils. Flavour and Fragrance Journal. 2008;23(4):213–26. https://onlinelibrary.wiley.com/doi/10.1002/ffj.1875
50. Instituto Nacional del Cáncer. Definición de antitumoral - Diccionario de cáncer del NCI - Instituto Nacional del Cáncer. https://www.cancer.gov/espanol/buscar/resultados?swKeyword=ANTITUMORAL
51. Harman D. Prolongation of life: role of free radical reactions in aging. J Am Geriatr Soc. 1969;17(8):721–35. https://doi.org/10.1111/j.1532-5415.1969.tb02286.x
52. Nádasi E, Varjas T, Pajor L, Ember I. Carcinogenic Potential ff Trans-2-hexenal Is Based on Epigenetic Effect. In Vivo (Brooklyn). 2005;19:559–62. Https://Iv.Iiarjournals.Org/Content/Invivo/19/3/559.Full.Pdf
53. Ulland BM, Weisburger JH, Yamamoto RS, Weisburger EK. Antioxidants and carcinogenesis: butylated hydroxytoluene, but not diphenyl-pphenylenediamine, inhibits cancer induction by N-2-fluorenylacetamide and by N-hydroxy-N-2-fluorenylacetamide in rats. Food Cosmet Toxicol. 1973;11(2):199–207. https://doi.org/10.1016/s0015-6264(73)80486-9
54. Matsushima T, Grantham PH, Weisburger EK, Weisburger JH. Phenobarbitalmediated increase in ring- and N-hydroxylation of the carcinogen N-2-fluorenylacetamide, and decrease in amounts bound to liver deoxyribonucleic acid. Biochem Pharmacol. 1972;21(15):2043–51. https://doi.org/10.1016/0006-2952(72)90158-X
55. Ohno Y, Takuma T, Asahi K ichi, Isono K. Differentiation induction of murine erythroleukemia cells by butylated hydroxytoluene. FEBS Letters. 1984;165(2):277–9. https://doi.org/10.1016/0014-5793(84)80185-4
56. Yamamoto Y, Inoue I, Takasaki T, Takahashi H. Inhibitory erects of selenium, vitamin A and butylated hydroxytoluene on growth of human maxillary cancer cells in vitro. Auris Nasus Larynx. 1996;23(1):91–97. https://doi.org/10.1016/S0385-8146(96)80014-9
57. Bakkali F, Averbeck S, Averbeck D, Idaomar M. Biological effects of essential oils – A review. Food and Chemical Toxicology. 2008;46(2):446–75. https://pubmed.ncbi.nlm.nih.gov/17996351/
58. Scala A, Allmann S, Mirabella R, Haring M, Schuurink R. Green Leaf Volatiles: A Plant’s Multifunctional Weapon against Herbivores and Pathogens. International Journal of Molecular Sciences. 2013;14(9):17781–811. https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3794753/
Downloads
Published
Issue
Section
License
Copyright (c) 2026 Eva Ruzena Montiel Fernández, Jorge L M. Ruíz
This work is licensed under a Creative Commons Attribution 4.0 International License.
