Phytochemical composition and cytotoxic assessment of Carica papaya leaf extracts on human squamous carcinoma-3 Cells (in vitro): a laboratory experimental
Abstract
Introduction: Oral cancer is one of the major diseases that can cause death worldwide. With the advancement of research on new drugs, medicinal plants have been explored as an alternative approach to cancer treatment. Papaya (Carica papaya) is a plant that has long been used in traditional medicine to treat various diseases. Ethanolic and aqueous extracts of C. papaya leaf have been reported to exhibit anticancer properties through mechanisms such as inhibiting cell proliferation and inducing apoptosis. This study aimed to analyze the cytotoxic effect of C. papaya leaf extract obtained with different extraction solvents on HSC-3 cell lines. Methods: This laboratory-based experimental study employed a post-test only control group design. The sample size was determined using the Federer formula, yielding four replicates per group. HSC-3 cell lines were treated with ethanolic and aqueous extracts of C. papaya leaf at concentrations of 125, 250, and 500 μg/mL for 24 hours, followed by a cytotoxicity test using the CCK-8 assay. Data were analyzed using one-way ANOVA followed by Tukey’s post hoc test. Results: Phytochemical analysis revealed the presence of alkaloids, flavonoids, and tannins in both ethanolic and aqueous extracts, whereas triterpenoids were detected only in the ethanolic extract, and saponins were detected only in aqueous extract. HSC-3 cells did not show any significant cytotoxic response to either extract at any tested concentration. Conclusion: Ethanolic and aqueous extracts of C. papaya leaf at concentrations of 125, 250, and 500 μg/mL did not exhibit any cytotoxic effects on HSC-3 cell lines.
Keywords
Full Text:
PDFReferences
Warnakulasuriya S, Filho, AM. Oral Cancer in the South and South-East Asia Region, 2022: Incidence and Mortality. Oral Diseases. 2025;31(5):1398-405.https://doi.org/10.1111/odi.15369.
Bray F, Laversanne, M, Sung, H, Ferlay, J, Siegel, RL, Soerjomataram, I, et al. Global cancer statistics 2022: GLOBOCAN estimates of incidence and mortality worldwide for 36 cancers in 185 countries. CA Cancer J Clin. 2024;74(3):229-63. https://doi.org/10.3322/caac.21834.
Gharat SA, Momin, MM, Bhavsar, C. Oral Squamous Cell Carcinoma: Current Treatment Strategies and Nanotechnology-Based Approaches for Prevention and Therapy. 2016;33(4):363-400. https://doi.org/10.1615/CritRevTherDrugCarrierSyst.2016016272.
Ding M, Yu, S, Chen, Y, Liu, Y, Xuan, F. Space-time analysis of head and neck cancer in Asia and its 34 countries and territories (1990-2021): Implications from the Global Burden of Disease Study 2021. PLoS One. 2025;20(6):e0326177. https://doi.org/10.1371/journal.pone.0326177.
Jayaraman S, Veeraraghavan, VP, Natarajan, SR, Jasmine, S. Exploring the therapeutic potential of curcumin in oral squamous cell carcinoma(HSC-3 cells): Molecular insights into hypoxia-mediated angiogenesis. Pathology - Research and Practice. 2024;254:155130. doi: https://doi.org/10.1016/j.prp.2024.155130.
Yang Z, Sun, P, Dahlstrom, KR, Gross, N, Li, G. Joint effect of human papillomavirus exposure, smoking and alcohol on risk of oral squamous cell carcinoma. BMC Cancer. 2023;23(1):457. https://doi.org/10.1186/s12885-023-10948-6.
Chu Y-H, Tzeng, S-L, Lin, C-W, Chien, M-H, Chen, M-K, Yang, S-F. Impacts of MicroRNA Gene Polymorphisms on the Susceptibility of Environmental Factors Leading to Carcinogenesis in Oral Cancer. PLOS ONE. 2012;7(6):e39777. https://doi.org/10.1371/journal.pone.0039777.
da Cruz Perez DE, Passos, KKM, Machado, RA, Martelli-Junior, H, Bonan, PRF. Continuing education in oral cancer during coronavirus disease 2019 (covid-19) outbreak. Oral Oncology. 2020;105:104713. https://doi.org/10.1016/j.oraloncology.2020.104713.
Manpreet K, Naveen Chandra, T, Seema, S, Arvind, K, Elena, ES. Ethnomedicinal Uses, Phytochemistry and Pharmacology of Carica papaya Plant: A Compendious Review. Mini-Reviews in Organic Chemistry. 2019;16(5):463-80. http://dx.doi.org/10.2174/1570193X15666180816110733.
Sharma A, Sharma, R, Sharma, M, Kumar, M, Barbhai, MD, Lorenzo, JM, et al. Carica papaya L. Leaves: Deciphering Its Antioxidant Bioactives, Biological Activities, Innovative Products, and Safety Aspects. Oxidative Medicine and Cellular Longevity. 2022;2022(1):2451733. doi: https://doi.org/10.1155/2022/2451733.
Nisa FZ, Astuti, M, Murdiati, A, Haryana, SM. Anti-proliferation and apoptosis induction of aqueous leaf extract of Carica papaya L. on human breast cancer cells MCF-7. Pakistan Journal of Biological Sciences. 2017;20(1):36-41. doi: https://doi.org/10.3923/pjbs.2017.36.41
Vuong QV, Hirun, S, Chuen, TLK, Goldsmith, CD, Murchie, S, Bowyer, MC, et al. Antioxidant and anticancer capacity of saponin-enriched Carica papaya leaf extracts. Int J of Food Science and Technology. 2015;50(1):169-77. https://doi.org/10.1111/ijfs.12618
Abd’quadri-Abojukoro AN, Nkadimeng, SM, McGaw, LJ, Nsahlai, IV. Phytochemical composition and cytotoxicity of ethanolic extracts of some selected plants. J of Applied Animal Research. 2022;50(1):656-65. https://doi.org/10.1080/09712119.2022.2119979.
Niyonizigiye I, Nkurunziza, D, Ngabire, D, Gitachew, AT, Chun, BS, Kim, G-D. Characterization and in vitro cytotoxicity of phytochemicals from Aspilia africana obtained using green extraction techniques. South African Journal of Botany. 2020;128:231-8. https://doi.org/10.1016/j.sajb.2019.11.013.
Mpingirika EZ, El Hosseiny, A, Bakheit, SMS, Arafeh, R, Amleh, A. Potential Anticancer Activity of Crude Ethanol, Ethyl Acetate, and Water Extracts of Ephedra foeminea on Human Osteosarcoma U2OS Cell Viability and Migration. BioMed Research International.2020;2020(1):3837693. https://doi.org/10.1155/2020/3837693.
Tessalonica S, Roeslan, MO. Effect of ethanol extracts from Persea americana leaves on HSC-3 proliferation. Journal of Indonesian Dental Association. 2020;3(2):65-70. https://doi.org/10.32793/jida.v3i2.582
Bozinou E, Pappas, IS, Patergiannakis, I-S, Chatzimitakos, T, Palaiogiannis, D, Athanasiadis, V, et al. Evaluation of Antioxidant, Antimicrobial, and Anticancer Properties of Onion Skin Extracts. 2023;15(15):11599. https://doi.org/10.3390/su151511599
Jawad M, Ali, M, Qasim, S, Akbar, A, Khan, NA, Sadiq, MB. Determination of Phenolic Compounds and Bioactive Potential of Plum (Prunus salicina) Peel Extract Obtained by Ultrasound-Assisted Extraction. BioMed Research International. 2022;2022(1):7787958. https://doi.org/10.1155/2022/7787958.
Karo-Karo SU, Arianto, A, Salim, E. Antibacterial Activity and Determination of Total Phenol and Flavonoid of Carica papaya L. Ethanol Extract. International Journal of Science, Technology & Management. 2023;4(1):233-8. https://doi.org/10.46729/ijstm.v4i1.738.
Mangkulion S, Roeslan, MO, Monthanpisut, P. The Potential of Cogon Grass (Imperata cylindrica) Ethanol Extract in Inhibiting Nitric Oxide Secretion in Fibroblast. Scientific Dental Journal. 2023;7(1).
Maheshwaran L, Nadarajah, L, Senadeera, SPNN, Ranaweera, CB, Chandana, AK, Pathirana, RN. Phytochemical Testing Methodologies and Principles for Preliminary Screening/ Qualitative Testing. Asian Plant Research Journal. 2024;12(5):11–38. https://doi.org/10.9734/aprj/2024/v12i5267.
Guediri I, Boubekri, C, Smara, O. Preliminary phytochemical screning from different extracts of Solanum nigrum plant growing in south of Algeria. Journal of Fundamental and Applied Sciences. 2023;12(2):624-33. https://doi.org/10.4314/jfas.v12i2.7.
Sudha R, Philip, XC, Suriyakumari, KVP. Phytochemical Constituents of Leaves of Moringa oleifera Grow in Cuddalore District, Tamil Nadu, India. SBV Journal of Basic, Clinical and Applied Health Science. 2020;3(4).
Lisa, Wa Ode, S, Suparman, S. Identification of Tannin Compounds In Ethanol Extract Of Cat's Whiskers Leaves (Orthosiphon aristatius) Using Thin Layer Chromatography. International Journal of Multidisciplinary Learners. 2025;1(3):106-10.
Roeslan MO, Tasha, G. Ethanol extract of Imperata Cylindrica leaves inhibits proliferation and migration of HSC-3 cell lines. Dent J (Majalah Kedokteran Gigi). 2021;54(3):150-4. https://doi.org/10.20473/j.djmkg.v54.i3.p150-154.
Kamiloglu S, Sari, G, Ozdal, T, Capanoglu, E. Guidelines for cell viability assays. Food Frontiers. 2020;1(3):332-49. https://doi.org/10.1002/fft2.44.
Ng ZX, Samsuri, SN, Yong, PH. The antioxidant index and chemometric analysis of tannin, flavonoid, and total phenolic extracted from medicinal plant foods with the solvents of different polarities. Journal of Food Processing and Preservation. 2020;44(9):e14680. doi: https://doi.org/10.1111/jfpp.14680.
Rokis A, Hanafi M, Ismail C, Haris MS, Lestari W, Ichwan S, et al. Medicinal plants in the Southeast Asia for treatment and prevention of oral cancer: A systematic review. Journal of Pharmacy & Pharmacognosy Research. 2025;13:1003-37.
Barnabas F, Wuyep P, Uchejeso O, Mantu E, Ukpa S, Nduke S, et al. Phytochemical analysis and antimicrobial effects of carica papaya leaf extracts on urinary tract infections.2023.
Ilham R, Lelo A, Harahap U, Widyawati T, Siahaan L. The Effectivity of Ethanolic Extract from Papaya Leaves (Carica papaya L.) as an Alternative Larvacide to Aedes spp. (1857-9655 (Print)).
Yuliani R, Syahdeni, F. Ethanolic Extract of Papaya Leaves (Carica papaya) and its Fractions have no Potential Cytotoxicity on T47D Cells. Pharmacon: Jurnal Farmasi Indonesia; Vol 17, No 1 (2020). 2020. https://doi.org/10.23917/pharmacon.v17i1.10760.
Kong YR, Jong, YX, Balakrishnan, M, Bok, ZK, Weng, JK, Tay, KC, et al. Beneficial Role of Carica papaya Extracts and Phytochemicals on Oxidative Stress and Related Diseases: A Mini Review. Biology. 2021 https://doi.org/10.3390/biology10040287.
Zhang D, Kanakkanthara, A. Beyond the Paclitaxel and Vinca Alkaloids: Next Generation of Plant-Derived Microtubule-Targeting Agents with Potential Anticancer Activity. Cancers. 2020 https://doi.org/10.3390/cancers12071721.
Kopustinskiene DM, Jakstas, V, Savickas, A, Bernatoniene, J. Flavonoids as Anticancer Agents. Nutrients. 2020 https://doi.org/ 10.3390/nu12020457.
Yi W, Yan, ZY, Shi, MQ, Zhang, Y, Liu, J, Yi, Q, et al. [Mechanism of tannins from Galla chinensis cream in promoting skin wound healing irats based on FAK/PI3K/Akt/mTOR signaling pathway]. Zhongguo Zhong Yao Za Zhi. 2025;50(2):480-97. https://doi.org/10.19540/j.cnki.cjcmm.20240902.706.
Ghante MH, Jamkhande, PG. Role of Pentacyclic Triterpenoids in Chemoprevention and Anticancer Treatment: An Overview on Targets and Underling Mechanisms. J Pharmacopuncture. 2019;22(2):55-67. https://doi.org/10.3831/KPI.201.22.007.
Chang L, Zhou, R, He, Y, Meng, M, Hu, J, Liu, Y, et al. Total saponins from Rhizoma Panacis Majoris inhibit proliferation, induce cell cycle arrest and apoptosis and influence MAPK signalling pathways on the colorectal cancer cell. Mol Med Rep. 2021;24(2):542. https://doi.org/10.3892/mmr.2021.12181.
Singh SP, Mishra, A, Shyanti, RK, Singh, RP, Acharya, A. Silver Nanoparticles Synthesized Using Carica papaya Leaf Extract (AgNPs-PLE) Causes Cell Cycle Arrest and Apoptosis in Human Prostate (DU145) Cancer Cells. Biological Trace Element Research. 2021;199(4):1316-31. https://doi.org/10.1007/s12011-020-02255-z.
Evans DM, Fang, J, Silvers, T, Delosh, R, Laudeman, J, Ogle, C, et al. Exposure time versus cytotoxicity for anticancer agents. Cancer Chemotherapy and Pharmacology. 2019;84(2):359-71. https://doi.org/10.1007/s00280-019-03863-w.
Munir S, Liu Z-W, Tariq T, Rabail R, Kowalczewski PŁ, Lewandowicz J, et al. Delving into the Therapeutic Potential of Carica papaya Leaf against Thrombocytopenia. Molecules [Internet]. 2022; 27(9).
Khor B-K, Chear NJ, Azizi J, Khaw K-Y. Chemical Composition, Antioxidant and Cytoprotective Potentials of Carica papaya Leaf Extracts: A Comparison of Supercritical Fluid and Conventional Extraction Methods. Molecules [Internet]. 2021; 26(5).
Mertens B, Nogueira, T, Stranska, R, Naesens, L, Andrei, G, Snoeck, R. Cidofovir is active against human papillomavirus positive and negative head and neck and cervical tumor cells by causing DNA damage as one of its working mechanisms. Oncotarget; Vol 7, No. 2016. doi: https://doi.org/10.18632/oncotarget.10100.
Pardenas I, Roeslan, M. Cytotoxicity Activity of Allium sativum Extracts Against HSC-3 cells. Quality improvement in dental and medical knowledge, research, skills and ethics facing global challenges. London: CRC Press; 2024. p. 393–8.
DOI: https://doi.org/10.24198/pjd.vol37no3.61396
Refbacks
- There are currently no refbacks.
All publications by the Universitas Padjadjaran [e-ISSN: 2549-6212, p-ISSN: 1979-0201] are licensed under a Creative Commons Attribution-ShareAlike 4.0 International License .





.png)
