Odovtos-International Journal of Dental Sciences (Odovtos-Int. J. Dent. Sc.), Online First, 2026. ISSN: 2215-3411

https://doi.org/10.15517/2096v718

https://revistas.ucr.ac.cr/index.php/rOdontos

BASIC RESEARCH:

Integrated Computational and Experimental Evaluation of Rutin as a Potential Therapeutic Agent for Oral Cancer

Evaluación computacional y experimental integrada de la rutina como agente terapéutico potencial para el cáncer oral

Madhan Shiva A.1 https://orcid.org/0009-0002-5393-7828

Monisha Prasad1 https://orcid.org/0000-0003-2185-8121

Manoj Kumar S.2 https://orcid.org/0009-0006-8367-3826

1Molecular Nutrition and Genomics Lab, Department of Community Medicine, Saveetha Medical College and Hospital, Saveetha Institute of Medical and Technical Sciences, Saveetha University, Chennai-602105, India.

2Translational Molecular Therapeutics and Nanotechnology Lab, Saveetha Medical College and Hospitals, Saveetha Institute of Medical and Technical Scien- ces (SIMATS), Saveetha University, Chennai, 602 105, Tamil Nadu, India.

Correspondence to: Manoj Kumar S. - manojkumars.smc@saveetha.com

Received: 25-II-2026 Accepted: 4-III-2026

ABSTRACT: Oral cancer remains a significant global health burden, requiring the development of effective and safer therapeutic options. Rutin, a naturally occurring flavonoid with antioxidant and anti-inflammatory properties, has gained attention for its potential anticancer activity. This study aimed to evaluate the therapeutic potential of rutin against oral cancer through a combined computational and experimental approach. Molecular docking was performed to assess rutin’s binding interactions with key molecular targets involved in oral cancer progression, particularly within the PI3K/AKT, MAPK, and NF-κB signaling pathways. Network and functional enrichment analyses were used to identify regulatory molecules and biological processes associated with rutin. Pharmacokinetic profiling was conducted to evaluate bioavailability and efficacy. In vitro assays were carried out to determine rutin’s effects on oral cancer cell viability, apoptosis induction, and caspase activation. Docking studies revealed strong interactions of rutin with critical proteins regulating cancer-related pathways. Network analysis highlighted EGFR, STAT3, and AP-1 as key nodes linked to apoptosis, cell cycle regulation, DNA repair, inflammation, and epithelial-mesenchymal transition. Pharmacokinetic predictions supported favorable absorption and bioavailability. In vitro findings demonstrated that rutin reduced oral cancer cell viability in a dose-dependent manner, enhanced apoptotic activity, and triggered caspase activation. Rutin exerts anticancer effects against oral cancer by modulating multiple signaling pathways and key regulatory molecules. These results provide mechanistic insights into its therapeutic potential and support further preclinical and clinical evaluation of rutin as a candidate for oral cancer treatment.

KEYWORDS: Rutin; Oral cancer; Molecular docking; Network analysis; Apoptosis; Cell viability.

RESUMEN: El cáncer oral sigue representando una carga significativa para la salud pública a nivel global, lo que subraya la necesidad de desarrollar terapias más eficaces y seguras. La rutina, un flavonoide natural con propiedades antioxidantes y antiinflamatorias, ha atraído la atención por su posible actividad anticancerígena. Este estudio tiene como objetivo evaluar el potencial terapéutico de la rutina en el tratamiento del cáncer oral mediante un enfoque combinado de métodos computacionales y experimentales. Se utilizó el acoplamiento molecular para estudiar las interacciones de unión de la rutina con dianas moleculares clave implicadas en la progresión del cáncer oral, en particular con las vías de señalización PI3K/AKT, MAPK y NF-κB. También se realizaron análisis de redes y de enriquecimiento funcional para identificar moléculas reguladoras y procesos biológicos asociados a la rutina. Además, se realizó un perfil farmacocinético para evaluar su biodisponibilidad y su eficacia. Los ensayos in vitro determinaron los efectos de la rutina sobre la viabilidad de las células cancerígenas orales, la inducción de apoptosis y la activación de caspasas. Los estudios de acoplamiento molecular mostraron interacciones fuertes entre la rutina y proteínas clave que regulan las vías relacionadas con el cáncer. El análisis de redes identificó a EGFR, STAT3 y AP-1 como nodos importantes asociados con la apoptosis, la regulación del ciclo celular, la reparación del ADN, la inflamación y la transición epitelio-mesénquima. Las predicciones farmacocinéticas indicaron una buena absorción y biodisponibilidad. Los resultados in vitro demostraron que la rutina redujo la viabilidad celular de forma dependiente de la dosis, promovió la apoptosis y activó las caspasas. En conjunto, la rutina muestra efectos anticancerígenos en el cáncer oral mediante la modulación de múltiples vías de señalización y de moléculas reguladoras clave, lo que respalda su potencial terapéutico en futuras investigaciones preclínicas y clínicas.

PALABRAS CLAVE: Rutina; Cáncer oral; Acoplamiento molecular; Análisis de redes; Apoptosis; Viabilidad celular.

INTRODUCTION

Oral cancer is among the ten most common cancers worldwide and remains a major contributor to morbidity and mortality (1). Despite significant advances in surgery, radiotherapy, hormonal therapy, and chemotherapy, outcomes are often unsatisfactory due to therapy resistance, recurrence, and debilitating side effects that compromise patient quality of life (2). This has created a strong interest in natural compounds with multitargeted anticancer potential and reduced toxicity. Flavonoids, a diverse class of polyphenolic compounds found in fruits, vegetables, and medicinal plants, are known for their antioxidant, anti-inflammatory, and anticancer effects (3). Rutin (quercetin-3-rutinoside), a flavonoid glycoside abundant in buckwheat, citrus fruits, and apples, has attracted attention for its wide range of pharmacological benefits, including its role in cancer prevention and therapy (4).

Preclinical studies suggest that rutin interferes with several hallmarks of oral cancer (5, 6). It suppresses uncontrolled cell proliferation, induces apoptosis, and inhibits invasion and metastasis. Mechanistically, rutin activates both intrinsic and extrinsic apoptotic pathways by modulating Bax, Bcl-2, and caspase activity (7). Fluorescence microscopy of rutin-treated oral cancer cells reveals apoptotic changes, including nuclear condensation and fragmentation (8).

Rutin also counteracts oxidative stress, a major driver of carcinogenesis (9). By scavenging reactive oxygen species (ROS), restoring glutathione, and enhancing antioxidant enzymes like superoxide dismutase and catalase, rutin reduces DNA damage and lipid peroxidation (10). Its anti-inflammatory effects further contribute to cancer suppression, as it inhibits TNF-α, IL-1β, IL-6, and NF-κB activation, thereby limiting chronic inflammation and tumor progression (11). Metastasis inhibition is another crucial function. Rutin downregulates matrix metalloproteinases (MMP-2 and MMP-9), reducing extracellular matrix degradation, and blocks VEGF-mediated angiogenesis, thus preventing tumor invasion and spread. At the molecular level, rutin modulates multiple oncogenic signaling pathways, including Ras/Raf/ERK, PI3K/Akt, MAPK, and TGF-β/Smad, highlighting its role as a multitarget therapeutic (12).

Moreover, rutin enhances the efficacy of conventional therapies by sensitizing cancer cells to chemotherapy and radiotherapy while potentially lowering drug toxicity. However, its clinical use faces challenges due to poor solubility and bioavailability. Nanotechnology-based formulations, such as rutin-loaded nanoparticles or cyclodextrin complexes, are being explored to overcome these limitations.

This study integrates molecular docking, ADME analysis, and in vitro assays to investigate rutin’s pharmacological potential against oral cancer. By combining computational modeling, network analysis, and experimental validation, we aim to clarify the molecular mechanisms underlying rutin’s anticancer activity and provide a foundation for its future development as a therapeutic candidate.

MATERIALS AND METHODS

Identification of differentially expressed genes (DEGs)

DEGs were identified from oral cancer datasets using GEO2R with cutoffs of p<0.01 and |logFC|>1. Oral cancer targets from GeneCards and rutin-related targets from CTD were compared, and overlaps were visualized using FunRich V3.1.3 with Venn diagrams.

Protein-protein interaction (PPI)

network and module analysis of DEGs in oral cancer

DEGs were mapped to the STRING database (v11.0) with a combined score>0.8 and visualized in Cytoscape (v3.8.2). Hub genes (degree>10) were identified, and key clusters were detected using MCODE and CytoHubba with standard parameters.

Gene ontology (GO) and pathway enrichment analysis

DEGs and gene clusters were analysed using g: Profiler for GO terms (MF, CC, BP) and KEGG pathways. Crosstalk analysis applied BH-adjusted p<0.05 with Jaccard and overlap coefficients>0.5, highlighting pathways relevant to rutin’s effects in oral cancer.

Molecular docking

Protein structures of key oral cancer targets were retrieved from the RCSB PDB and preprocessed by removing water molecules, adding hydrogens, and correcting structural irregularities using Chimera or PyMOL. The rutin ligand (PubChem CID: 5280805) was prepared by adding hydrogens, optimizing geometry, and converting to PDBQT format using OpenBabel or MarvinS-ketch. Molecular docking was performed with AutoDock Vina, in which grid boxes were defined around the active sites of target proteins. Binding affinities and poses were analysed based on docking scores and key interactions (hydrogen bonds, hydrophobic contacts, van der Waals forces), providing mechanistic insights into rutin's therapeutic potential against oral cancer.

Cell line maintenance

KB oral cancer cells are obtained from NCCS, Pune and grown in T25 culture flasks using Dulbecco's Modified Eagle's Medium (DMEM) supplemented with 10% fetal bovine serum (FBS) and 1% antibiotics (usually penicillin) streptomycin). This substrate composition supports cell growth by providing essential nutrients and maintaining sterility through antibiotics. Cells are incubated at 37°C in a humidified atmosphere containing 5% CO2, which are optimal conditions for their growth and metabolism. Regular passage of cells occurs after confluence to prevent overgrowth and maintain cell health.

MTT cell viability assay

To assess the cytotoxic effects of rutin, KB cells are plated in 96-well plates at a density of 5x103 cells per well. Before processing, cells are starved in serum-free medium for 3 h to synchronise their metabolic state and reduce possible interference from serum factors. Different concentrations of rutin are then applied to the cells, which are incubated for 24 h. After the treatment period, an MTT assay is performed by adding MTT reagent to each well, which is converted into formazan crystals by living cells. The formazan crystals are dissolved and the absorbance is measured using a microplate reader at 570 nm. Cell viability is calculated using the following formula: % cell viability=[A570 nm treated cells/A570 nm control cells]×100 (13).

Morphological study

After determining the MTT, the optimal routine dose with a significant cytotoxic effect is selected. KB-1 cells are seeded in 6-well plates at a density of 2x105 cells per well and treated with this optimal dose for 24 h. After treatment, cells are gently washed with phosphate-buffered saline (PBS) to remove excess rutin and observed under a phase-contrast microscope. Changes in cell morphology such as cell shrinkage, membrane blebbing and detachment are observed, providing qualitative insight into the apoptotic effects induced by rutin.

Cell imaging by AO/EtBr staining

To further characterise the induction of apoptosis, 24 h routine-treated KB cells are collected, washed with ice-cold PBS, and stained with acridine orange. (AO) and ethidium bromide (EtBr). AO stains live cells green, while EtBr stains apoptotic cells red due to reduced cell membrane integrity. Stained cells are visualized under a fluorescence microscope to detect and quantify apoptotic changes, providing further confirmation of the cytotoxic effect of rutin on KB cells.

Statistical analysis

All experimental data were analysed using one-way ANOVA followed by Student’s t-test. Results are presented as mean ± SD of triplicates, with p<0.05 considered statistically significant.

RESULTS

Identification of DEGs in HCC

A total of 322 and 198 DEGs were identified from GSE3524 and GSE23558, respectively, using GEO2R with thresholds of adjusted p<0.01 and logFC>1. Volcano plots indicated predominant upregulation in oral cancer samples. Intersection analysis in FunRich V3.1.3 revealed 105 common upregulated DEGs between the two datasets, as shown in the Venn diagram and expression patterns Figure 1.

Figure 1. Intersecting targets of CTD & Genecard.

Identification of DEGs involved in pterostilbene in oral cancer

Venn analysis identified 23 overlapping targets between rutin and oral cancer from GeneCards, CTD, and GSE datasets. STITCH (confidence >0.4) and Cytoscape were used to visualise networks, separating 112 upregulated and 86 downregulated DEGs. The upregulated network comprised 24 nodes and 112 edges, with CytoHubba identifying top-ranking hub genes. These core genes and modules were further analysed via GO and pathway analysis to elucidate their roles in oral cancer (Figure 2).

Figure 2. Hub genes.

Identification of GO and KEGG pathway enrichment analysis of upregulated genes

GO analysis of Profiler revealed that upregulated DEGs in oral cancer were enriched in biological processes such as positive regulation of epithelial-to-mesenchymal transition, inflammatory response, and negative regulation of apoptosis, with molecular functions including IL-1 receptor, cadherin, and protein kinase binding, and cellular components like the catenin and adherens junction complexes. KEGG pathway analysis showed significant enrichment in the Wnt signalling pathway, general cancer pathways, and oral cancer-specific pathways. These results highlight key molecular mechanisms influenced by rutin, providing insights for targeted therapeutic interventions (Figure 3).

Docking analysis

Molecular docking of rutin with key apoptosis-related proteins revealed binding affinities ranging from –3.89 to –5.96 kcal/mol. Rutin showed notable interactions with BCL2 (–5.61 kcal/mol) through hydrogen bonds with THR7, ASN11, GLN190, TRP195, and ARG6, involving residues MET16, ASN11, and VAL16. Binding to TP53 (– 4.61 kcal/mol) involved THR153, ARG306, and GLU336, GLY302, LYS120 residues, while interaction with BAX (–5.96 kcal/mol) occurred via GLN32 with ARG94, VAL95, ALA96, TRP188, THR186, and THR182. MCL-1 showed weaker binding (–3.89 kcal/mol) through THR191 and LYS279. These results indicate that rutin forms stable interactions with multiple apoptotic proteins, suggesting its potential modulatory role in oral cancer pathways (Table 1 and Figure 4).

Figure 3. Gene Ontology & KEGG enrichment analysis.

Table 1. Molecular docking analysis rutin with targeted proteins.

S.No	Name of the Protein	Binding Affinity Value	Hydrogen Interaction	Amino acid residues
1	BCL2 with Rutin	-5.61	THR 7, ASN11-HSN 1, ASN11-HD21 1, GLN190-HE22 1, TRP195-HE1 1,	MET16A, ASN11, VAL16
2	TP53with Rutin	-4.61	THR153-HN,ARG306-HH12,ARG306-HH22,UNK0-H	GLU336,GLY302,LYS120
3	BAX with Rutin	-5.96	GLN32	ARG94,VAL95,ALA96,TRP188,THR186,THR182
4	MCL-1	-3.89	UNK0:H, UNK0:H, UNK0:H	THR191,LYS279

Figure 4. 2D& 3D interactions of rutin with targets.

MTT

The cytotoxic effect of Rutin on oral cancer cells was evaluated using the MTT assay. A dose-dependent reduction in cell viability was observed with increasing concentrations of Rutin (25-200 µM/ml) (Figure 5). Compared to the control group, which showed 100% viability, Rutin treatment significantly decreased cell survival, with viability dropping to approximately 25% at the highest tested concentration (200 µM/ml). The IC50 value was calculated as 124.03 ± 4.82 µM/ml, indicating the effective inhibitory concentration of Rutin on oral cancer cell proliferation.

Morphological changes

Phase-contrast microscopic examination revealed that KB oral cancer cells in the control group retained normal morphology, displaying an elongated and adherent structure with intact cell membranes. In contrast, cells treated with Rutin (124 µM/ml) exhibited prominent morphological alterations indicative of cytotoxicity and apoptosis, such as cell shrinkage, rounding, detachment from the culture surface, and increased loss of cell density. These changes confirm the growth-inhibitory and apoptotic effect of Rutin at IC50 concentration (Figure 6).

AO/EtBr staining

Fluorescence microscopy analysis of AO/EtBr dual-stained KB oral cancer cells demonstrated clear apoptotic features in Rutin (124 µM/ml)-treated cells compared to the control (Figure 7). Control cells exhibited normal green fluorescence with intact nuclei, while treated cells showed significant apoptotic characteristics such as nuclear condensation and fragmentation. Quantitative analysis revealed a markedly higher percentage of apoptotic cells in the Rutin-treated group at IC50 concentration (55%) compared to the control (5%), confirming the pro-apoptotic effect of Rutin in oral cancer cells.

Figure 5. Effect of Rutin on oral cancer cell viability assessed by MTT assay. Cells were treated with increasing concentrations of Rutin (25-200 µM/ml) for 24 h. A dose-dependent decline in cell viability was observed, with an IC50 value of 124.03 ± 4.82 µM/ml. Data are presented as mean ± SEM of three independent experiments. p<0.05 vs. control.

Figure 6. Morphological changes in KB oral cancer cells after Rutin treatment. Phase-contrast microscopy images show untreated control cells with normal elongated morphology compared to Rutin (124 µM/ml)-treated cells, which exhibit apoptotic features including shrinkage, rounding, and detachment from the surface.

Figure 7. Detection of apoptotic cells in Rutin (124 µM/ml)-treated KB oral cancer cells by AO/EtBr dual staining. The histogram shows the percentage of apoptotic cells in the control group (green bar) and Rutin-treated group at IC50 concentration (red bar). A significant increase in apoptosis was observed in Rutin-treated cells compared to the control. Values are expressed as mean ± SEM.

DISCUSSION

Network analysis of rutin-associated signalling pathways in oral cancer represents a pivotal exploration into its therapeutic potential. Rutin, renowned for its anti-inflammatory and antioxidant properties, has garnered attention for its anticancer effects (14). This study employs advanced bioinformatics tools and extensive molecular databases to construct and analyse comprehensive pharmacological networks. By elucidating complex signalling pathways, gene expression profiles, and protein-protein interactions (PPIs) affected by rutin, researchers gain a detailed understanding of its role in oral cancer treatment (15). The investigation begins with the identification of differentially expressed genes (DEGs) from oral cancer datasets using robust analytical tools like GEO2R and Limma (16). These DEGs are further validated through intersection analysis, highlighting their involvement in critical biological processes and molecular functions within oral cancer (17). The construction of PPI networks using databases such as STRING and visualisation with Cytoscape aids in pinpointing hub genes and key regulatory nodes influenced by rutin (18).

Functional enrichment analyses, including GO and KEGG pathway analyses, provide deeper insights into how rutin modulates pathways crucial to cancer progression (19). Notably, rutin impacts pathways related to apoptosis, cell proliferation, and metastasis inhibition, which are pivotal in oral tumorigenesis (20). The study identifies significant associations between rutin and key signalling pathways in oral cancer, such as the Wnt signalling pathway and pathways in cancer, underscoring its potential as a therapeutic agent. Molecular docking simulations revealed favourable binding interactions between rutin and selected target proteins involved in critical cellular processes like apoptosis and cell proliferation. The identification of hydrogen bonds, hydrophobic contacts, and van der Waals forces in the protein-ligand complexes underscores rutin's potential to modulate these pathways, potentially inhibiting cancer cell growth and promoting apoptosis . Such interactions are pivotal for designing novel therapeutic strategies that target specific molecular mechanisms underlying oral cancer progression.

This study evaluating the anticancer effects of Rutin in an oral cancer cell line using in vitro cell viability and apoptosis assays provides important results and implications. Rutin, a flavonoid known for its antioxidant and anti-inflammatory properties, showed significant effects in this study. The results show a dose-dependent decrease in cell viability, suggesting the potential of Rutin as a cytotoxic agent against oral cancer cells.

In addition, the induction of apoptosis observed in treated cells further supports the therapeutic potential of Rutin. This program-mediated cell death mechanism is crucial in cancer treatment and underscores Rutin's ability to trigger cancer cell elimination pathways without significant cytotoxicity to normal cells (21). However, the study also reveals some limitations and areas for further research. The need for in vivo studies to confirm these results in a more complex biological environment is obvious. In addition, investigating the molecular mechanisms underlying Rutin's anticancer effects would provide deeper insights into its modes of action and potential synergy with existing therapies.

CONCLUSION

This study provides a comprehensive computational framework for evaluating rutin as a potential therapeutic agent against oral cancer. By integrating molecular docking, rutin was shown to form stable interactions with key cancer-related proteins, highlighting its ability to modulate molecular networks and pathways involved in tumor progression.These findings underscore the importance of bioinformatics in identifying novel targets and elucidating complex molecular mechanisms, laying the foundation for experimental validation and future clinical studies. Overall, rutin demonstrates promising anticancer properties, supporting its potential as a natural compound for adjuvant or alternative therapies in oral cancer treatment.

ETHICAL: Not applicable.

AUTHOR CONTRIBUTION STATEMENT: Conceptualization and design: M.S.A., M.P., and M.K.S.; Literature review: M.S.A. and M.P.; Methodology and validation: M.S.A. and M.K.S.; Formal analysis: M.S.A.; Investigation and data collection: M.S.A. and M.P.; Resources: M.K.S.; Data analysis and interpretation: M.S.A. and M.K.S.; Writing-original draft preparation: M.S.A.; Writing-review & editing: M.P. and M.K.S.; Funding acquisition: M.K.S.

References

1. Awinashe M., Viswaja K., Pathath A.W., Subair F.P., Varshney A., Mishra P., Mulla M., Mulla M. CRISPR-cas9-Mediated Gene Editing to Reverse Oncogenic Mutations in Oral Squamous Cell Carcinoma. Annals of African Medicine:10-4103.

2. Zafar A., Khatoon S., Khan M.J., Abu J., Naeem A. Advancements and limitations in traditional anti-cancer therapies: a comprehensive review of surgery, chemotherapy, radiation therapy, and hormonal therapy. Discover oncology. 2025 Apr 24; 16 (1): 607.

3. Bustonova S., Alijonov M. Biologically Active Substances of Medicinal Plants and Their Benefits. Journal of Applied Science and Social Science. 2025 Mar 11;1 (1): 12-8.

4. Soosai D., Ramalingam R., Perumal E., Veeramani K., Pancras C., Almutairi M.H., Savarimuthu L.A., Veeramuthu D., Antony S. Anticancer effects of rutin from Fagopyrum tataricum (tartary buckwheat) against osteosarcoma cell line. Molecular biology reports. 2024 Dec; 51 (1): 312.

5. Nouri Z., Fakhri S., Nouri K., Wallace C.E., Farzaei M.H., Bishayee A. Targeting multiple signaling pathways in cancer: The rutin therapeutic approach. Cancers. 2020 Aug 14; 12 (8): 2276.

6. Butnariu M., Quispe C., Sharifi-Ra J., Pons-Fuste E., Lopez-Jorne P., Zam W., Das T., Dey A., Kumar M., Pentea M., Eid A.H. Naturally-occurring bioactives in oral cancer: Preclinical and clinical studies, bottlenecks and future directions.

7. Pandey P., Khan F., Qari H.A., Oves M. Rutin (Bioflavonoid) as cell signaling pathway modulator: Prospects in treatment and chemoprevention. Pharmaceuticals. 2021 Oct 22; 14 (11): 1069.

8. Pandey P., Khan F., Alzahrani F.A., Qari H.A., Oves M. A novel approach to unraveling the apoptotic potential of rutin (bioflavonoid) via targeting Jab1 in cervical cancer cells. Molecules. 2021 Sep 12; 26 (18): 5529.

9. Imani A., Maleki N., Bohlouli S., Kouhsoltani M., Sharifi S., Maleki Dizaj S. Molecular mechanisms of anticancer effect of rutin. Phytotherapy Research. 2021 May; 35 (5): 2500-13.

10. Enogieru A.B., Haylett W., Hiss D.C., Bardien S., Ekpo O.E. Rutin as a potent antioxidant: Implications for neurodegenerative disorders. Oxidative medicine and cellular longevity. 2018; 2018 (1): 6241017.

11. Farha A.K., Gan R.Y., Li H.B., Wu D.T., Atanasov A.G., Gul K., Zhang J.R., Yang Q.Q., Corke H. The anticancer potential of the dietary polyphenol rutin: Current status, challenges, and perspectives. Critical Reviews in Food Science and Nutrition. 2022 Jan 25; 62 (3): 832-59.

12. Nagoor Meeran M.F., Goyal S.N., Suchal K., Sharma C., Patil C.R., Ojha S.K. Pharmacological properties, molecular mechanisms, and pharmaceutical development of asiatic acid: a pentacyclic triterpenoid of therapeutic promise. Frontiers in pharmacology. 2018 Sep 4; 9: 892.

13. Supino R. MTT assays. InIn vitro toxicity testing protocols 1995 Jan 1 (pp. 137-149). Totowa, NJ: Humana Press.

14. Farha A.K., Gan R.Y., Li H.B., Wu D.T., Atanasov A.G., Gul K., Zhang J.R., Yang Q.Q., Corke H. The anticancer potential of the dietary polyphenol rutin: Current status, challenges, and perspectives. Critical Reviews in Food Science and Nutrition. 2022 Jan 25; 62 (3): 832-59.

15. Ahmad A.V., Ruhi M., Ali S.A. Unraveling the therapeutic mechanisms of myristic acid and luteolin 7-rutinoside in oral cancer: insights from network pharmacology and molecular docking analysis. Pharmacol Discov. 2024; 4 (2): 7.

16. Abbas S.Z., Qadir M.I., Muhammad S.A. Systems-level differential gene expression analysis reveals new genetic variants of oral cancer. Scientific reports. 2020 Sep 4; 10 (1): 14667.

17. Rout T., Mohapatra A., Kar M. A systematic review of graph-based explorations of PPI networks: methods, resources, and best practices. Network Modeling Analysis in Health Informatics and Bioinformatics. 2024 May 30; 13 (1): 29.

18. Kumar R., Samal S.K., Routray S., Dash R., Dixit A. Identification of oral cancer related candidate genes by integrating protein-protein interactions, gene ontology, pathway analysis and immunohistochemistry. Scientific Reports. 2017 May 30; 7 (1): 2472.

19. Gatasheh M.K. Identifying key genes against rutin on human colorectal cancer cells via ROS pathway by integrated bioinformatic analysis and experimental validation. Computational Biology and Chemistry. 2024 Oct 1;112: 108178.

20. Nouri Z., Fakhri S., Nouri K., Wallace C.E., Farzaei M.H., Bishayee A. Targeting multiple signaling pathways in cancer: The rutin therapeutic approach. Cancers. 2020 Aug 14; 12 (8): 2276.

21. Muthukumaran M., Edwardpaul F.R., Abdulbari N., Bupesh G., Kirubakaran D. Rhizophora mucronata Flower Extracts as a Source of Anticancer and Antimicrobial Agents: GC-MS, FTIR and Molecular Docking Approaches. Pharmacological Research-Natural Products. 2025 Dec 2: 100445.