Investigation of In Vitro Cytotoxicity of Hypericum Perforatum in Pancreatic Cancer

Öz

Pancreatic cancer remains one of the most aggressive malignancies with poor prognosis and limited treatment options. In view of the constraints imposed by current treatment modalities, there has been an increased focus on natural products as potential complementary agents in cancer therapy. Hypericum perforatum (HP), commonly known as St. John’s Wort, has demonstrated cytotoxic and apoptotic properties in various cancer models; however, its effects on pancreatic cancer cells are not well-studied. This study aimed to investigate the in vitro cytotoxic effects of Hypericum perforatum extract on AR42J pancreatic cancer cells. AR42J cells were cultured and treated with increasing concentrations (3.125–100 µg/mL) of HP extract for 24 and 48 hours. Cell viability was assessed using the MTT assay. Nuclear morphological changes were evaluated by DAPI staining and visualized with a Juli™ Smart Fluorescent Cell Analyzer. HP extract exhibited a dose- and time-dependent cytotoxic effect on AR42J cells. IC₅₀ values were determined to be 12.5 µg/mL at 24 hours and 6.25 µg/mL at 48 hours. DAPI staining confirmed nuclear condensation and fragmentation in treated cells, supporting apoptosis as a mechanism of action. These results indicate that H. perforatum extract effectively reduced pancreatic cancer cell viability while inducing apoptotic cell death. These findings suggest that H.perforatum extract exerts significant antiproliferative and pro-apoptotic effects on AR42J pancreatic cancer cells. The study highlights its potential as a plant- based complementary therapeutic candidate for pancreatic cancer treatment.

Anahtar Kelimeler

• Pancreatic cancer
• Hypericum Perforatum
• in vitro study
• AR42J cell
• MTT assay
• DAPI staining

Kaynakça

1. [1]. Akbulut, S. 2018. Application of photolon guided photodynamic therapy in pancreatic cancer treatment. Istanbul University Institute of Science and Technology Biomedical Engineering Program. (In Turkish)
2. [2]. Sun, L., Shang, H., Wu, Y., & Xin, X. 2022.Hypericin-mediated photodynamic therapy enhances gemcitabine-induced Capan-2 cell apoptosis via inhibiting NADPH level. Journal of Pharmacy and Pharmacology, 74, 596–604.
3. [3]. Tanker, M., & Tanker, N. 1985. Herba Hyperici, Hypericum Perforatum. Pharmacognosy I. Ankara University, Faculty of Pharmacy, No:58, 177–179.
4. [4]. Tokur, S. 1988. Studies on the ecology of some Hypericum species. Natura Turk Botanical Journal. (In Turkish)
5. [5]. Schneider, M., & Marquard, R. 1996. Aufnahme und Akkumulation von Cadmium und weiterer Schwermetalle bei Hypericum Perforatum L. und Linum Usitatissimum L. Z. Arznei- und Gewürzpflanzen ;1, 111-116.
6. [6]. Baytop, T. 1999. Treatment with plants in Turkey (Past and present). Istanbul University Faculty of Pharmacy, Istanbul, 166-167.
7. [7]. Kamuhabwa, A. R., Agostinis, P., D’Hallewin, M. A., Kasran, A., & De Witte, P. A. 2000. Photodynamic activity of hypericin in human urinary bladder carcinoma cells. Anticancer Research, 20, 2579-2584.
8. [8]. Schempp, C. M., Kirkin, V., Simon-Haarhaus, B., Kersten, A., Kiss, J., Termeer, C. C., Gilb, B., Kaufmann, T., Borner, C., Sleeman, J. P., & Simon, J. C.2002. Inhibition of tumour cell growth by hyperforin, a novel anticancer drug from St. John’s Wort that acts by induction of apoptosis. Oncogene, 21, 1242–1250.

9. [9]. Kaçar, O., & Azkan, N. 2004. Relationships between hypericin and top drog herb yield and some morphological and agronomic characteristics in St. John's Wort (Hypericum Perforatum L.). Uludag University, Faculty of Agriculture Journal, 18(2), 109-122. (In Turkish)
10. [10]. Stephens, L. L. 2004. An investigation into the antidepressant activity of Hypericum Perforatum. Rhodes University, A Thesis Submitted in Fulfillment of the Requirements for the Degree of Doctor of Philosophy in Pharmacy.
11. [11]. Mutlubaş, H., & Özdemir, Z. 2020. Applications of Hypericum Perforatum in traditional medicine. Kırklareli University, Institute of Science, University of Health Sciences, Hamidiye Faculty of Pharmacy. Integrative and Anatolian Medical Journal, 1(3). (In Turkish)
12. [12]. Menegazzi, M., Masiello, P., & Novelli, M. 2021. Anti-tumor activity of Hypericum Perforatum L. and hyperforin. Antioxidants, 10(1), 18. https://doi.org/10.3390/antiox10010018
13. [13]. Ekren, S., Sonmez, C., & Bayram, E. 2011. Determination of some agricultural and quality traits in Hypericum Perforatum L. clones. Journal of Agricultural Sciences.
14. [14]. Ayhan, F., & Altınkaynak, D. 2020. Hypericum Perforatum feasibility report and investor guide. Ministry of Agriculture and Forestry, Education and Publication Department, Ankara. (In Turkish)
15. [15]. Çirak, C., & Kurt, D. 2014.Hypericum species and their uses as important medicinal plants. Anatolia, Journal of AARI, 24(1), 38-52. (In Turkish)
16. [16]. Gonenc, T. M., Ozturk, M., Turkseven, S. G., Kırmızıbayrak, P. B., Günal, S., & Yılmaz, 2020. S. Hypericum Perforatum L.: An overview of the anticancer potencies of the specimens collected from different ecological environments. Pakistan Journal of Botany, 52(3), 1003-1010.
17. [17]. Gaedcke, F., 2003.Beispiel Johanniskraut. Pharm Unserer Zeit, 32, 192-201.
18. [18]. Linde, K. (2008). Hypericum Perforatum L for depression - An overview of randomized studies based on individual extracts. Quality and Transparency of Phytopharmaceuticals Commission of the Research, Natural Medicine Committee.
19. [19]. Hostettmann, K., & Wolfender, J. L. 2005. St. John’s Wort and its active principles in depression and anxiety. Phytochemistry, 5-20.
20. [20]. Ersoy, E., Ozkan, E., & Mat, A. 2019. Pharmacological activities of Hypericum species in the light of new studies. Istanbul University Journal of Advanced Research in Health Sciences, 2(2).
21. [21]. Barnes, J., Anderson, L. A., & Philipson, D. 2001. St John’s Wort (Hypericum Perforatum L.): A review of its chemistry, pharmacology and clinical properties. Journal of Pharmacy and Pharmacology, 53, 583–600.
22. [23]. Shrivastava, M., & Dwivedi, L. K. 2015. Therapeutic potential of Hypericum Perforatum: A review. International Journal of Pharmaceutical Sciences and Research, 6(12), 1000-1007.
23. [24]. Jung, J. H., Hwang, J., Kim, J. H., Sim, D. Y., Im, E., Park, J. E., Park, W. Y., Shim, B. S., Kim, B., & Kim, S. H. 2021. Phytochemical candidates repurposing for cancer therapy and their molecular mechanisms. Seminars in Cancer Biology, 68, 164-174.
24. [25]. Çakmak, H. E., & Bayram, E. 2003. Determination of some agronomic and quality characteristics of St. Hypericum Perforatum L. populations from Muğla origin. Aegean University Faculty of Agriculture Journal, 40(1), 57-64.
25. [26]. Hu, Z., Zhao, P., & Xu, H. 2020.Hyperoside exhibits anticancer activity in non-small cell lung cancer cells with T790M mutations by upregulating Foxo1 via CCAT1. Oncology Reports, 43, 617-624.
26. [27]. Candoken, E., Kuruca, S. E., & Akev, N. 2017.Evaluation of the anticancer effects of Aloe Vera and Aloe-Emodin on B16F10 murine melanoma and NIH3T3 mouse embryogenic fibroblast cells. Istanbul Journal of Pharmacy, 47(3), 77-83.
27. [28]. Ghazanfari, T., Yaraee, R., Rahmati, B., Hakimzadeh, H., Shams, J., & Nadoushan, M. R. J.2011. In vitro cytotoxic effect of garlic extract on malignant and nonmalignant cell lines. Immunopharmacology and Immunotoxicology, 33(4), 603–608.
28. [29]. Sengupta, A., Ghosh, S., & Das, S., 2004.Modulatory influence of garlic and tomato on cyclooxygenase-2 activity, cell proliferation, and apoptosis during azoxymethane-induced colon carcinogenesis in rat. Cancer Letters, 208, 127–136.
29. [30]. Vlavcheski, F., O’Neill, E. J., Gagacev, F., & Tsiani, E. 2022. Effects of berberine against pancreatitis and pancreatic cancer. Molecules, 27, 8630.