Carvacrol is a Novel Natural Therapeutic Approach Through the Inhibition of Proliferation, Autophagy and Migration in Pancreatic Ductal Adenocarcinoma Cells

Abstract

Objective: Pancreatic ductal adenocarcinoma, which is the most common and aggressive pancreatic cancer, has the highest mortality rate of cancers because of difficulties in diagnosis and chemoresistance. As the chemotherapeutic options are very limited and insufficient for PDAC, novel effective therapeutic approaches are urgently needed for pancreatic cancer patients. Carvacrol naturally found in thyme (Thymus vulgaris), wild bergamot (Citrus aurantium var. bergamia Loisel), black cumin (Nigella sativa), marjoram (Origanum scabrum, Origanum microphyllum, Origanum onites, Origanum vulgare) and black pepper (Lepidium flavum) plants is shown to have antibacterial and antioxidant effects. In this study, we aimed to investigate the anticarcinogenic potential of carvacrol through proliferation and autophagy in PDAC cells. Material and Method: To determine the anti-proliferative effects of carvacrol in Panc-1 cells, we performed the MTS assay using different carvacrol doses of 100, 200, 300, 400, 500, 600, 700 and 800 μM at 24h, 48h, 72h. The gene and protein expressions of Atg16L1 and Beclin-1, autophagy key mediators, were analyzed by RT-PCR and western blot in Panc-1 cells treated with 300 and 400 μM for 24h, 48h. Additionally, the migrative property of PDAC cells was evaluated using a wound healing assay. Results: Based on MTS results, carvacrol significantly inhibited cell proliferation in the doses of 300 and 400 μM at 24h and 48 h in Panc-1. These same doses led to decreased autophagy and migration through Atg16L1, and Beclin-1 expressions. Conclusion: Our findings first revealed that carvacrol has promising value as a potential therapeutic approach for PDAC. We believe that further mechanistic investigations will be a guide for its clinical usage.

Keywords

• Pancreatic cancer
• carvacrol
• proliferation
• autophagy
• migration

Supporting Institution

None.

Thanks

None.

References

1. 1. Sim W, Lim WM, Hii LW, Leong CO, Mai CW. Targeting pancreatic cancer immune evasion by inhibiting histone deacetylases. World J Gastroenterol 2022;14;28(18):1934–45.
2. 2. Pourshams A, Sepanlou SG, Ikuta KS, Bisignano C, Safiri S, Roshandel G, et al. The global, regional, and national burden of pancreatic cancer and its attributable risk factors in 195 countries and territories, 1990–2017: A systematic analysis for the Global Burden of Disease Study 2017. Lancet Gastroenterol Hepatol 2019;4(12):934–47.
3. 3. Atkinson MA, Campbell-Thompson M, Kusmartseva I, Kaestner KH. Organisation of the human pancreas in health and diabetes. Diabetologia 2020;7;63(10):1966–73.
4. 4. Mihaljevic AL, Michalski CW, Friess H, Kleeff J. Molecular mechanism of pancreatic cancer—understanding proliferation, invasion, and metastasis. Langenbecks Arch Surg 2010;18;395(4):295–308.
5. 5. Stefanoudakis D, Frountzas M, Schizas D, Michalopoulos NV., Drakaki A, Toutouzas KG. Significance of TP53, CDKN2A, SMAD4 and KRAS in pancreatic cancer. Curr Issues Mol Biol 2024;23;46(4):2827–44.
6. 6. Sönmez EH; Gürbüz N. Pankreatik duktal adenokarsinoa hücrelerinde miRNA’nın, otofaji üzerinde düzenleyici etkisinin değerlendirilmesi. Süleyman Demirel Üniversitesi Tıbbi Biyoloji Anabilim Dalı Tezi. Isparta: Süleyman Demirel Üniversitesi. 2018.
7. 7. Turantepe E; Gürbüz N. Flavopiridol’ün pankreas kanseri hücrelerinde anti-proliferatif etkilerinin değerlendirilmesi. 2024. 8. Dalby KN, Tekedereli I, Lopez-Berestein G, Ozpolat B. Targeting the prodeath and prosurvival functions of autophagy as novel therapeutic strategies in cancer. Autophagy 2010;6(3):322–9.
8. 9. Ashour AA, Abdel-Aziz AAH, Mansour AM, Alpay SN, Huo L, Ozpolat B. Targeting elongation factor-2 kinase (eEF-2K) induces apoptosis in human pancreatic cancer cells. Apoptosis 2014;19(1):241–58.

9. 10. Li J, Chen X, Kang R, Zeh H, Klionsky DJ, Tang D. Regulation and function of autophagy in pancreatic cancer. Autophagy. 2021;17(11):3275–96.
10. 11. Grant TJ, Hua K, Singh A. Molecular Pathogenesis of Pancreatic Cancer. In 2016. p. 241–75.
11. 12. Abdel Hadi N, Reyes-Castellanos G, Carrier A. Targeting redox metabolism in pancreatic cancer. Int J Mol Sci 2021;22(4):1534.
12. 13. Koçatakan P, Ataseven H. Pankreas kanseri. Ankara Eğitim ve Araştırma Hastanesi Tıp Dergisi. 2021;27;54(1):59–65.
13. 14. He X, Wang N, Zhang Y, Huang X, Wang Y. The therapeutic potential of natural products for treating pancreatic cancer. Front Pharmacol 2022;2:13.
14. 15. Yıldız Ş, Turan S. Timokinon, Timol ve Karvakrolün antioksidan aktiviteleri ve lipit oksidasyonunu önleme kapasiteleri. Atatürk Üniversitesi Ziraat Fakültesi Dergisi 2021;52(1):108-118.
15. 16. Sharifi‐Rad M, Varoni EM, Iriti M, Martorell M, Setzer WN, del Mar Contreras M, et al. Carvacrol and human health: A comprehensive review. Phytotherapy Research 2018;32(9):1675–87.
16. 17. Suntres ZE, Coccimiglio J, Alipour M. The Bioactivity and toxicological actions of carvacrol. Crit Rev Food Sci Nutr 2015;55(3):304–18.
17. 18. Srinivasan MK, Premnath BJ, Parimelazhagan R, Namasivayam N. Synthesis, characterization, and evaluation of the anticancer properties of pH‐responsive carvacrol‐zinc oxide quantum dots on breast cancer cell line (MDA‐MB‐231). Cell Biochem Func. 2024;42(4).
18. 19. Abed AT, Almudhafar AMH, Hadi NR. Anti-Cancer study of carvacrol in hypoxic-ınduced colorectal cancer cell. Journal of Angiotherapy 2024;8(1). 20. Ahmad A, Abbas Al-Fatlawi A. Cytotoxicity and pro-apoptotic activity of carvacrol on human breast cancer cell line MCF-7. Available from: http://www.wjpsonline.org/
19. 21. Luo Y, Wu JY, Lu MH, Shi Z, Na N, Di JM. Carvacrol Alleviates Prostate Cancer Cell Proliferation, Migration, and Invasion through Regulation of PI3K/Akt and MAPK Signaling Pathways. Oxid Med Cell Longev 2016;10(1).
20. 22. Bansal A, Saleh-E-In MdM, Kar P, Roy A, Sharma NR. Synthesis of carvacrol derivatives as potential new anticancer agent against lung cancer. Molecules 2022;19;27(14):4597.
21. 23. Dai W, Sun C, Huang S, Zhou Q. Carvacrol suppresses proliferation and invasion in human oral squamous cell carcinoma. Onco Targets Ther 2016;2297.
22. 24. Yin Q hua, Yan F xiang, Zu XY, Wu Y hua, Wu X ping, Liao M chu, et al. Anti-proliferative and pro-apoptotic effect of carvacrol on human hepatocellular carcinoma cell line HepG-2. Cytotechnology. 2012;64(1):43–51.
23. 25. Kılıç Y, Geyikoglu F, Çolak S, Turkez H, Bakır M, Hsseinigouzdagani M. Carvacrol modulates oxidative stress and decreases cell injury in pancreas of rats with acute pancreatitis. Cytotechnology 2016;68(4):1243–56.
24. 26. Eroglu Gunes C, Secme M, Kurar E, Donmez H. Apoptotic and Anti-Metastatic Effect of Carvacrol in PANC-1 Human Pancreatic Cancer Cells. Natural Products and Biotechnology 2022;2.