Potential anticancer effects of Peganum harmala in human papillomavirus-related cervical and head and neck cancer cells

Hiba Faroug Muddather¹Tohfa Nasibova^2,3Gábor J. Szebeni^4,5Nikolett Gémes⁵Noémi Bózsity¹Renáta Minorics¹Eldar Garayev³ILKAY ERDOGAN ORHAN⁶Gaëtan Herbette^7,8Zsuzsanna Schelz^1*Judit Hohmann^2,9István Zupkó^10,11*

• ¹Institute of Pharmacodynamics and Biopharmacy, University of Szeged, Szeged, Hungary
• ²Institute of Pharmacognosy, University of Szeged, Szeged, Hungary
• ³Department of General and Toxicological Chemistry, Azerbaijan Medical University, Baku, Azerbaijan
• ⁴Department of Internal Medicine, Hematology Centre, University of Szeged, Szeged, Hungary
• ⁵Laboratory of Functional Genomics, Institute of Genetics, HUN-REN Szegedi Biologiai Kutatokozpont, Szeged, Hungary
• ⁶Department of Pharmacognosy, Lokman Hekim Universitesi, Ankara, Türkiye
• ⁷Institut de Chimie des Substances Naturelles, Universite Paris-Saclay, Gif-sur-Yvette, France
• ⁸Spectropole, FSCM, Centrale Marseille, CNRS, Aix-Marseille Universite, Marseille, France
• ⁹HUN-REN–USZ Biologically Active Natural Products Research Group, Szegedi Tudomanyegyetem, Szeged, Hungary
• ¹⁰Interdisciplinary Centre of Natural Products, University of Szeged, Szeged, Hungary
• ¹¹Institute of Pharmacodynamics and Biopharmacy, Szegedi Tudomanyegyetem, Szeged, Hungary

Background: Peganum harmala L (P. harmala) has a longstanding role in ethnomedical treatments. Its reported anticancer effects have led to increased research interest; however, its antineoplastic properties on cervical and head and neck (HN) cancer cells need further investigation. In this study, we investigated P. harmala''s harmala's antineoplastic effects on HPV-infected cervical and HN cancer cell lines. Methods: The crude extract derived from multiple plant parts and isolated β-carboline alkaloids was tested on several human neoplastic cell types using the MTT-based analysis. Apoptosis was examined by fluorescent double staining, Annexin V-Alexa 488–propidium iodide labeling, and caspase-3 assays. Moreover, flow cytometry was employed to explore the cell cycle progression alterations, while the tubulin polymerization assay assessed influences on microtubule dynamics. The antimetastatic property was investigated by wound healing and transwell invasion assays to explore the impact on cellular motility and invasiveness, respectively. Results: The IC50 values were calculated and examined relative to non-malignant fibroblasts to assess selective toxicity. The root extract demonstrated the most substantial growth inhibitorygrowth-inhibitory effect among the tested extracts. Harmine, one of the isolated bioactive alkaloids, showed a substantial effect, with inhibitory concentrations between 6.05 and 27.85 µM. Apoptosis induced by harmine was confirmed through cellular morphological appearance, flow cytometric evaluations, and caspase-3 activation. Evaluation Assessment of the cell cycle demonstrated that harmine disrupted cell cycle progression, particularly increasing the apoptotic sub-G1 population and G2/M phase populations. Moreover, it revealed the ability to stabilize microtubules. Our findings showed that harmine and the root extract significantly reduced cell migration. Furthermore, harmine was found to have anti-invasive properties. Conclusion: These findings showed potential harmine antiproliferative and antimetastatic activities, indicating its potential for further research in developing natural therapeutic agents.