Evaluation of the Effect of Thymol on the Cytotoxicity of Cetuximab in Lung Cancer Cells

Öz

The treatment of lung cancer continues to be a significant challenge for many oncologists and their patients. Treatment using epidermal growth factor receptor inhibitors is connected to a positive outcome. Cetuximab, a chimeric monoclonal antibody targeting the epidermal growth factor receptor (EGFR), in conjunction with monoterpene phenol thymol, is recommended for the treatment of lung cancer. While a mild acne-like skin rash is quite frequent in patients using cetuximab, a severe rash is rare. The goal of the current study was to assess whether thymol could enhance the anticancer effectiveness of cetuximab in A-549, non-small cell lung cancer (NSCLC) cell line. We found that the combination of cetuximab and thymol synergistically suppressed cell proliferation by inducing membrane damaging, oxidative stress, and apoptosis in A-549 cells. Taken together, our results indicate that the combination of thymol and cetuximab could improve anticancer responses and may notably enhance treatment outcomes in NSCLC.

Anahtar Kelimeler

• Cetuximab
• carvacrol
• lung cancer
• synergistic anticancer effect
• apoptosis

Timol’ün Akciğer Kanseri Hücrelerinde Cetuximab'ın Sitotoksisitesi Üzerindeki Etkisinin Değerlendirilmesi

Öz

Akciğer kanserinin tedavisi, birçok onkolog ve hasta için önemli bir zorluk olmaya devam etmektedir. Epidermal büyüme faktörü reseptörü inhibitörleri kullanarak yapılan tedaviden olumlu bir sonuçlar alınmaktadır. Epidermal büyüme faktörü reseptörünü (EGFR) hedefleyen bir kimerik monoklonal antikor olan cetuximab, monoterpen fenol timol ile birlikte, akciğer kanserinin tedavisi için önerilmektedir. Cetuximab kullanan hastalarda hafif akne benzeri cilt döküntüsü oldukça sık görülürken, şiddetli döküntü nadir görülmektedir. Mevcut çalışmanın amacı, timolün cetuximabın A-549, küçük hücreli olmayan akciğer kanseri (NSCLC) hücre dizisindeki antikanser etkinliğini artırıp artırmayacağını değerlendirmektir. Cetuximab ve timol kombinasyonunun A-549 hücrelerinde zar hasarını, oksidatif stresi ve apoptozu uyararak hücre proliferasyonunu sinerjik bir şekilde baskıladığını bulduk. Bir arada değerlendirildiğinde, bulgularımız timol ve cetuximab kombinasyonunun antikanser yanıtların iyileştirilebileceğini ve NSCLC'deki tedavi sonuçlarının önemli ölçüde arttırılabileceğini göstermektedir.

Anahtar Kelimeler

• Cetuximab
• karvakrol
• akciğer kanseri
• sinerjik antikancer etki
• apoptoz

Kaynakça

1. Alar, T., & Şahin, E.M. (2012). Akciğer kanseri: birinci basamakta tanı, tedavi ve korunma Lung cancer: Diagnosis, treatment and prevention in primary care. Smyrna Tıp Dergisi, 68-74.
2. Albanell, J., Rojo, F., Averbuch, S., Feyereislova, A., Mascaro, J.M., Herbst, R., … & Baselga, J. (2002). Pharmacodynamic studies of the epidermal growth factor receptor inhibitor ZD1839 in skin from cancer patients: histopathologic and molecular consequences of receptor inhibition. Journal of Clinical Oncology, 20, 110-124. https://doi.org/10.1200/jco.20.1.110
3. Alnuqaydan, A.M., Rah, B., Almutary, A.G., & Chauhan, S.S. (2020). Synergistic antitumor effect of 5-fluorouracil and withaferin-A induces endoplasmic reticulum stress-mediated autophagy and apoptosis in colorectal cancer cells. American Journal of Cancer Research, 10(3), 799.
4. Ambrož, M., Matoušková, P., Skarka, A., Zajdlová, M., Žáková, K., & Skálová, L. (2017). The effects of selected sesquiterpenes from myrica rubra essential oil on the efficacy of doxorubicin in sensitive and resistant cancer cell lines. Molecules, 22(6), 1021. https://doi.org/10.3390/molecules22061021
5. Anitha, A., Maya, S., Sivaram, A.J., Mony, U., & Jayakumar, R. (2016). Combinatorial nanomedicines for colon cancer therapy. Wiley Interdisciplinary Reviews: Nanomedicine and Nanobiotechnology, 8, 151-159. https://doi.org/10.1002/wnan.1353
6. Balta, B.Z., Üre, Ö.S., Erturan, S., & Aydin, G. (2013). Prognostic factors in advanced-stage non-small cell lung cancer. Haseki Tip Bulteni, 51(2), 56-60. https://doi.org/10.4274/Haseki.848
7. Bonner, J.A., Harari, P.M., Giralt, J., Azarnia, N., Shin, D.M., Cohen, R.B., … & Ang, K.K. (2006). Radiotherapy plus cetuximab for squamous-cell carcinoma of the head and neck. New England Journal of Medicine, 354, 567-578. https://doi.org/10.1056/NEJMoa053422
8. Bradford, M.M. (1976). A rapid and sensitive method for the quantization of microgram quantities of protein utilizing the principle of protein dye binding. Analytical Biochemistry, 72, 248-254. https://doi.org/10.1006/abio.1976.9999

9. Cai, W.Q., Zeng, L.S., Wang, L. ., Wang, Y.Y., Cheng, J.T., Zhang, Y., … & Xin, H.W. (2020). The latest battles between EGFR monoclonal antibodies and resistant tumor cells. Frontiers in Oncology, 10, 1249. https://doi.org/10.3389/fonc.2020.01249
10. Cao, S., Xia, M., Mao, Y., Zhang, Q., Donkor, P.O., Qiu, F., & Kang, N. (2016). Combined oridonin with cetuximab treatment shows synergistic anticancer effects on laryngeal squamous cell carcinoma: involvement of inhibition of EGFR and activation of reactive oxygen species-mediated JNK pathway. International Journal of Oncology, 49(5), 2075-2087. https://doi.org/10.3892/ijo.2016.3696
11. Ceylan, K.C., Arpat, A.H., & Kaya, Ş.Ö. (2009). Küçük hücreli dışı akciğer kanserinde klinik ve patolojik evrelemenin karşılaştırılması. Dr. Suat Seren Göğüs Hastalıkları ve Cerrahisi Eğitim Araştırma Hastanesi, Göğüs Cerrahisi, 14.
12. Chung, K.Y., Shia, J., Kemeny, N.E., Shah, M., Schwartz, G.K., Tse, A., … & Saltz, L.B. (2005). Cetuximab shows activity in colorectal cancer patients with tumors that do not express the epidermal growth factor receptor by immunohistochemistry. Journal of Clinical Oncology, 23, 1803-1810. https://doi.org/10.1200/jco.2005.08.037
13. Ding, L., Getz, G., Wheeler, D.A., Mardis, E.R., McLellan, M.D., Cibulskis, K., … & Wilson, R.K. (2008). Somatic mutations affect key pathways in lung adenocarcinoma. Nature, 455(7216), 1069-1075. https://doi.org/10.1038/nature07423
14. Ergelen, R., & Çimşit, N.Ç. (2000). Akciğer tümörü. Türk Toraks Derneği, 178-188. https://doi.org/10.5152/tcb.2013.30
15. Flohe, L., & Gunzler, W.A. (1984). Glutathione peroxidase. Methods in Enzymology, 105, 115-121. https://doi.org/10.1016/S0076-6879(84)05015-1
16. Huang, F., Wu, X.N., Chen, J.I.E., Wang, W.X., & Lu, Z.F. (2014). Resveratrol reverses multidrug resistance in human breast cancer doxorubicin-resistant cells. Experimental and Therapeutic Medicine, 7, 1611-1616. https://doi.org/10.3892/etm.2014.1662
17. Işıtmangil, G.A. (2013). Akciğer kanser immünoterapı̇sı̇nde yenı̇ stratejı̇ler. Haydarpaşa Numune Eğitim ve Araştırma Hastanesi Tıp Dergisi, 53(3), 168-177.
18. Jacot, W., Bessis, D., Jorda, E., Ychou, M., Fabbro, M., Pujol, J. L., & Guillot, B. (2004). Acneiform eruption induced by epidermal growth factor receptor inhibitors in patients with solid tumours. British Journal of Dermatology, 151(1), 238-241. https://doi.org/10.1111/j.1365-2133.2004.06026.x
19. Javed, H., Azimullah, S., Meeran, M.F., Ansari, S.A., & Ojha, S. (2019). Neuroprotective effects of thymol, a dietary monoterpene against dopaminergic neurodegeneration in rotenone-induced rat model of Parkinson’s disease. International Journal of Molecular Sciences, 20(7), 1538. https://doi.org/10.3390/ijms20071538
20. Kaja, S., Payne, A.J., Naumchuk, Y., & Koulen, P. (2017). Quantification of lactate dehydrogenase for cell viability testing using cell lines and primary cultured astrocytes. Current Protocols in Toxicology, 72, 2-26. https://doi.org/10.1002/cptx.21
21. Lacouture, M.E. (2006). Mechanisms of cutaneous toxicities to EGFR inhibitors. Nature Reviews Cancer, 6(10), 803-812. https://doi.org/10.1038/nrc1970
22. Lacouture, M.E., Basti, S., Patel, J., & Benson, A.3rd (2006). The SERIES clinic: an interdisciplinary approach to the management of toxicities of EGFR inhibitors. The Journal of Supportive Oncology, 4(5), 236-238.
23. Leeman-Neill, R.J., Wheeler, S.E., Singh, S.V., Thomas, S.M., Seethala, R.R., Neill, D.B., … & Grandis, J.R. (2009). Guggulsterone enhances head and neck cancer therapies via inhibition of signal transducer and activator of transcription-3. Carcinogenesis, 30, 1848-1856. https://doi.org/10.1093/carcin/bgp211
24. McComb, S., Chan, P. ., Guinot, A., Hartmannsdottir, H., Jenni, S., Dobay, M.P., Bourquin, J.P., & Bornhauser, B.C. (2019). Efficient apoptosis requires feedback amplification of upstream apoptotic signals by effector caspase-3 or-7. Science Advances, 5, eaau9433. https://doi.org/10.1126/sciadv.aau9433
25. Mosmann, T. (1983). Rapid colorimetric assay for cellular growth and survival: Application to proliferation and cytotoxicity assays. Journal of Immunological Methods, 65, 55-63. https://doi.org/10.1016/0022-1759(83)90303-4
26. Nagoor Meeran, M.F., & Stanely Mainzen Prince, P. (2012). Protective effects of thymol on altered plasma lipid peroxidation and nonenzymic antioxidants in isoproterenol induced myocardial infarcted rats. Journal of Biochemical and Molecular Toxicology, 26, 368-373. https://doi.org/10.1002/jbt.21431
27. Nagoor Meeran, M.F., Javed, H., Al Taee, H., Azimullah, S., & Ojha, S.K. (2017). Pharmacological properties and molecular mechanisms of thymol: prospects for its therapeutic potential and pharmaceutical development. Frontiers in pharmacology, 8, 380. https://doi.org/10.3389/fphar.2017.00380
28. Parhamifar, L., Andersen, H., & Moghimi, S.M. (2019). Lactate dehydrogenase assay for assessment of polycation cytotoxicity. Nanotechnology for Nucleic Acid Delivery: Methods and Protocols, 291-299. https://doi.org/10.1007/978-1-4939-9092-4_18
29. Park, S.J., Kim, M.J., Kim, Y.K., Kim, S.M., Park, J.Y., & Myoung, H. (2010). Combined cetuximab and genistein treatment shows additive anti-cancer effect on oral squamous cell carcinoma. Cancer Letters, 292(1), 54-63. https://doi.org/10.1016/j.canlet.2009.11.004
30. Parkin, D.M., Bray, F., Ferlay, J., & Pisani, P. (2005). Global cancer statistics. CA: A Cancer Journal for Clinicians, 55(2), 74-108. http://doi.org/10.3322/canjclin.55.2.74
31. Pei, J., Pan, X., Wei, G., & Hua, Y. (2023). Research progress of glutathione peroxidase family (GPX) in redoxidation. Frontiers in Pharmacology, 14, 1147414. https://doi.org/10.3389/fphar.2023.1147414
32. Peréz-Soler, R., & Saltz, L. (2005). Cutaneous adverse effects with HER1/EGFR-targeted agents: is there a silver lining?. Journal of Clinical Oncology, 23(22), 5235-5246. https://doi.org/10.1200/jco.2005.00.6916
33. Üstüner, C., & Entok, E. (2019). Experimental animal models for lung cancer. Nuclear Medicine Seminars, 5, 40-48. https://doi.org/10.4274/nts.galenos.2019.0006
34. Zhang, L., Li, X.F., Tand, L., Zhao, Y.M., & Zhou, C.C. (2014). The effects of cetuximab in combination with docetaxel for the acquired resistance to EGFR-TKI in non-small cell lung cancer cells. Tumor, 34, 584-590.