Anticancer Activity of Opuntia Cochenillifera Extract Against Triple-Negative Breast Cancer Cells via Apoptosis Induction and Bioinformatics-Based Target Prediction
Sažetak
Background/Aim: Breast cancer remains one of the most commonly diagnosed cancers among women worldwide and is a significant public health concern. Aim of this study was to investigate the in vitro anticancer activity of Opuntia cochenillifera ethanolic extract (EOC) against triple‑negative breast cancer (TNBC) cells and to predict potential molecular targets involved in its mechanism of action through bioinformatics analysis.
Methods: Fresh O cochenillifera leaves were extracted by ethanol maceration. Phytochemical constituents were analysed using qualitative screening, thin‑layer chromatography (TLC), UV–Vis spectrophotometry and Fourier transform infrared (FTIR) spectroscopy. Cytotoxicity against MDA‑MB‑231 TNBC cells was determined via MTT assay and morphological assessment. Apoptosis induction was quantified by Annexin V–FITC/PI flow cytometry. Potential molecular targets were identified through integrated bioinformatics platforms, including STITCH, SEA, SwissTargetPrediction, STRING and Cytoscape, followed by pathway enrichment analysis.
Results: Phytochemical analysis revealed the presence of alkaloids, flavonoids, phenolics, saponins and steroids, with flavonoids as predominant constituents (68 mg/g extract). FTIR spectra confirmed hydroxyl, aliphatic, aromatic and carbonyl functional groups consistent with phenolic and flavonoid structures. EOC exhibited dose‑dependent cytotoxicity against MDA‑MB‑231 cells, with an IC₅₀ of 270.63 µg/mL, accompanied by morphological and flow‑cytometric evidence of apoptosis. Network pharmacology analysis identified 34 apoptosis‑related target genes, including six hub genes (CDKN1A/p21, TP53, MAPK1, MAPK3, AKT1 and FOXO3), associated with TP53‑p21 signalling, MAPK cascades and mitochondrial stress‑induced apoptosis pathways.
Conclusion: O cochenillifera ethanolic extract demonstrated significant cytotoxic and pro‑apoptotic effects in TNBC cells, potentially mediated through TP53‑p21 and MAPK pathway modulation. These findings suggest that O cochenillifera is a promising candidate for further investigation as a natural adjuvant therapeutic agent for triple‑negative breast cancer.
Reference
Xiong L, Lu H, Hu Y, Wang W, Liu R, Wan X, et al. In vitro anti-motile effects of rhoifolin, a flavonoid extracted from Callicarpa nudiflora on breast cancer cells via downregulating podocalyxin-ezrin interaction during epithelial mesenchymal transition. Phytomedicine. 2021;1–30. doi: 10.1016/j.phymed.2021.153486.
Severo P, Ferreira C, Luan C, Passos A, Lima J, Fialho E. Effect of quercetin and chrysin and its association on viability and cell cycle progression in MDA-MB-231 and MCF-7 human breast cancer cells. Biomed Pharmacother. 2024;179:1–9. doi: 10.1016/j.biopha.2024.117276
Suzery M, Cahyono B, Amalina ND. Antiproliferative and apoptosis effect of hyptolide from Hyptis pectinata (L.) Poit on human breast cancer cells. J Appl Pharm Sci. 2020 Feb;10(2):1–6. doi: 10.7324/JAPS.2020.102001.
Zhang J, Wu Y, Li Y, Li S, Liu J, Yang X, et al. Natural products and derivatives for breast cancer treatment: from drug discovery to molecular mechanism. Phytomedicine. 2024;129:1–24. doi: 10.1016/j.phymed.2024.155600.
Hu D, Li Y, Li R, Wang M, Zhou K. Recent advances in reactive oxygen species (ROS)-responsive drug delivery systems for photodynamic therapy of cancer. Acta Pharm Sin B. 2024;:1–11. doi: 10.1016/j.apsb.2024.10.015.
Helen H, Carensia M, Halim P, Riza M, Ahmed A, Dalimunthe A, et al. Flavonoids as modulators of miRNA expression in pancreatic cancer: pathways, mechanisms, and therapeutic potential. Biomed Pharmacother. 2024;179:1–23. doi: 10.1016/j.biopha.2024.117347.
Setiani S, Sulasmi S, Arumingtyas EL. Plant diversity and utilization of traditional home garden in Bangkalan District, Madura Island, Indonesia. Russ J Agric Socio-Econ Sci. 2022;2(122):93–102. doi: 10.18551/rjoas.2022-02.11.
Pérez-Viveros DJ, Díaz-Batalla L, Navarro-Cortez RO, Palma-Rodríguez HM, Hernández JP. Effect of extrusion on physicochemical and functional properties of cladode flour from Opuntia cochenillifera and Opuntia ficus-indica. Appl Food Res. 2024;4:100479. doi: 10.1016/j.afres.2024.100479.
Vachetta VS, Marder M, Troncoso MF, Elola MT. Opportunities, obstacles and current challenges of flavonoids for luminal and triple-negative breast cancer therapy. Eur J Med Chem Rep. 2022;6(8):1–26. doi: 10.1016/j.ejmcr.2022.100077.
Li Y, Zhang X, Wang Y, Li J, Li Y, Zhang Y, et al. High-efficiency, flexibility and lead-free X-ray shielding multilayered polymer composites: experiment and simulation study. Sci Rep. 2021;11:4279. doi: 10.1038/s41598-021-83852-7.
Hermansyah D, Munir D, Lelo A, Putra A, Amalina ND, Alif I. The synergistic antitumor effects of Curcuma longa and Phyllanthus niruri extracts on promoting apoptotic pathways in breast cancer stem cells. Thai J Pharm Sci. 2022;46(5):541–50. doi: 10.56808/3027-7922.2638.
Amalina ND, Suzey M, Cahyono B. Cytotoxic activity of Hyptis pectinata extracts on MCF-7 human breast cancer cells. Indones J Cancer Chemoprevention. 2020 Feb;2:1–6. Doi:10.3524/IJCCIndonesia2019.
Dirja BT, Wardoyo EH, Putra A, Amalina ND. Enhanced third degree burn wound healing by hypoxic mesenchymal stem cells’ secretome: IL-10 upregulation and TNF-α/PGE2 suppression. Trends Sci. 2024; 21(10):8288. doi: 10.48048/tis.2024.8288.
Cahyono B, Suzery M, Amalina ND, Bima DN. Synthesis and antibacterial activity of epoxide from hyptolide (Hyptis pectinata (L.) Poit) against Gram-positive and Gram-negative bacteria. J Appl Pharm Sci. 2020 Dec;10(12):13–22. doi: 10.7324/JAPS.2020.101202.
Amalina N, Nurhayati IP, Meiyanto E. Doxorubicin induces lamellipodia formation and cell migration. Indones J Cancer Chemoprevention. 2017;8(2):61–7. doi: 10.14499/indonesianjcanchemoprev8iss2pp61-67.
Amalina ND, Salsabila IA, Zulfin UM, Jenie RI, Meiyanto E. In vitro synergistic effect of hesperidin and doxorubicin downregulates epithelial-mesenchymal transition in highly metastatic breast cancer cells. J Egypt Natl Canc Inst. 2023;35(1):–. doi: 10.1186/s43046-023-00166-3.
Jenie RI, Amalina ND, Hermawan A, Suzery M, Putra A, Meiyanto E. Caesalpinia sappan reduces the stemness of breast cancer stem cells involving the elevation of intracellular reactive oxygen species. Res Pharm Sci. 2023;18:25. doi: 10.4103/1735-5362.389959.
Mursiti S, Amalina ND, Marianti A. Inhibition of breast cancer cell development using Citrus maxima extract through increasing levels of reactive oxygen species (ROS). J Phys Conf Ser. 2021;1918(5):052005. doi: 10.1088/1742-6596/1918/5/052005.
Yang Z, Zhang Y, Liu Z, Zhang Q, Liu C, Chen X, et al. Identification of key genes in HER2-positive breast cancer with brain metastasis using bioinformatics analysis. Transl Cancer Res. 2023;12(5):1045–56. doi: 10.21037/tcr-22-2752.
Amalina ND, Suzery M, Cahyono B, Jenie RI, Hermawan A, Meiyanto E. [Revealing the potential of secondary metabolites of Indonesian herbal plants to stop breast cancer metastasis: an in-silico approach]. Indones J Cancer Chemoprevention. 2020;11(3):150–60. doi: 10.14499/indonesianjcanchemoprev11iss3pp150-160. Indonesian.
Zeghbib W, Boudjouan F, Vasconcelos V, Lopes G. Phenolic compounds’ occurrence in Opuntia species and their role in the inflammatory process: a review. Molecules. 2022;27(15):4763. doi: 10.3390/molecules27154763.
Queral-Beltran A, Lacorte S, Marin-Garcia M, Tauler R. UV-Vis absorption spectrophotometry and LC-DAD-MS-ESI(+)-ESI(-) coupled to chemometrics analysis of the monitoring of sulfamethoxazole degradation by chlorination, photodegradation, and chlorination/photodegradation. Anal Chim Acta. 2023;1276(7):1–14. doi: 10.1016/j.aca.2023.341563.
Alqasas N, Johnson D. Determination of Hansen solubility parameters of water-soluble proteins using UV–Vis spectrophotometry. Heliyon. 2023 Nov;9(11):e21403. doi: 10.1016/j.heliyon.2023.e21403.
Ocampo F, No E, Tatiana E, Andr A. Drug loading comparison of commercial ibuprofen on magnetite nanoparticles surface by UV–Vis spectrophotometry and acid-alkali titration by a factorial design of experiments. OpenNano. 2023;14(10):1–10. doi: 10.1016/j.onano.2023.100193.
Liu Y, Wang J, Li X, Zhang H, Chen L, Li J, et al. Study on the protective effect of flavonoids extracted from Jatropha curcas leaves against radiation damage. Heliyon. 2024;10(20):e39256. doi: 10.1016/j.heliyon.2024.e39256.
Sari DP, Santoso A, Wijaya CH. Cytotoxic test of ethanol extract of bintaro fruit and peel (Cerbera odollam Gaertn.) against Artemia salina Leach larvae using BSLT method. Int J Curr Sci Res Rev. 2023;7(7):2345–52. doi: 10.47191/ijcsrr/V7-i07-18.
Pratiwi R, Sari LK, Nugroho AE. Phytochemical characterization and flavonoid identification in Codiaeum variegatum leaf extract. J Pharm Sci Community. 2022;9(2):45–52. doi: 10.24071/jpsic.2022.2.45.
Matos TKB, Guedes JAC, Filho EGA, Luz LR, Lopes GS, Nascimento RF, et al. Integrated UPLC-HRMS, chemometric tools, and metabolomic analysis of forage palm (Opuntia spp. and Nopalea spp.) to define biomarkers associated with non-susceptibility to carmine cochineal (Dactylopius opuntiae). J Braz Chem Soc. 2021;32(11):2201–14. doi: 10.21577/0103-5053.20210345.
Tungmunnithum D, Pinthong D, Hano C. Medicinal plant: uses in traditional medicine, phytochemistry and pharmacological activities. Medicines (Basel). 2018;5(4):127. doi: 10.3390/medicines5040127.
Suzery M, Cahyono B, Amalina ND. Citrus sinensis (L.) peels extract inhibits metastasis of breast cancer cells by targeting the downregulation matrix metalloproteinases-9. Open Access Maced J Med Sci. 2021;9(B):464–9. doi: 10.3889/oamjms.2021.6072.
Amalina ND, Wahyuni S, Harjito. Cytotoxic effects of the synthesized Citrus aurantium peels extract nanoparticles against MDA-MB-231 breast cancer cells. J Phys Conf Ser. 2021;1918(3):032006. doi: 10.1088/1742-6596/1918/3/032006.
Paramita DA, Hermansyah D, Amalina ND. Regulation of p53 and survivin by Curcuma longa extract to caspase-3 dependent apoptosis in triple negative breast cancer cells. Med Glas (Zenica). 2022;19(2):189–96. doi: 10.17392/1453-22.
Hermansyah D, Putra A, Munir D, Lelo A, Amalina ND, Alif I. Synergistic effect of Curcuma longa extract in combination with Phyllanthus niruri extract in regulating Annexin A2, epidermal growth factor receptor, matrix metalloproteinases, and pyruvate kinase M1/2 signaling pathway on breast cancer stem cell. J Pharm Res Int. 2021;9:271–85. doi: 10.3889/oamjms.2021.5941.
Suzery M, Cahyono B, Amalina ND. Unveiling the potential of Hyptis capitata and Hyptis brevipes compounds in overcoming apoptosis resistance and inducing targeted cell cycle arrest in breast cancer stem cells. Trends Sci. 2024;21(12):–. doi: 10.48048/tis.2024.8404.
Cahyono B, Amalina ND, Suzery M, Bima DN. Exploring the capability of Indonesia natural medicine secondary metabolite as potential inhibitors of SARS-CoV-2 proteins to prevent virulence of COVID-19: in silico and bioinformatic approach. Open Access Maced J Med Sci. 2021;9:336–42. doi: 10.3889/oamjms.2021.5945.
- Authors retain copyright and grant the journal right of first publication with the work simultaneously licensed under a Creative Commons Attribution License that allows others to share the work with an acknowledgement of the work's authorship and initial publication in this journal.
- Authors are able to enter into separate, additional contractual arrangements for the non-exclusive distribution of the journal's published version of the work (e.g., post it to an institutional repository or publish it in a book), with an acknowledgement of its initial publication in this journal.
- Authors are permitted and encouraged to post their work online (e.g., in institutional repositories or on their website) prior to and during the submission process, as it can lead to productive exchanges, as well as earlier and greater citation of published work (See The Effect of Open Access).