The effects of cetuximab with agomelatine on gene expression in colon cancer cells

Abstract

This study investigated the combined effects of agomelatine, a melatonergic antidepressant, and cetuximab, an EGFR inhibitor, on the colorectal cancer cell line (Caco-2). Caco-2 cells were treated with agomelatine (0.3 μg/ml and 3 μg/ml) and cetuximab (50 μg/ml), individually and in combination, for 24 and 48 hours. Cell viability was assessed using the MTT assay. Gene expression analysis of EGFR, BCL2, PIK3CA, BAX, mTOR, and AKT3 was performed using real-time PCR. All treatment groups showed significant decreases in cell viability compared to the control (p<0.05), with enhanced effects in combined treatments. EGFR expression was significantly reduced in drug-treated groups, particularly with cetuximab (p<0.05). While changes were observed in BCL2, PIK3CA, BAX, mTOR, and AKT3 expression, these were not statistically significant (p>0.05). This study demonstrates the potential synergistic cytotoxic effects of agomelatine and cetuximab on Caco-2 colorectal cancer cells. The significant reduction in EGFR expression suggests a potential mechanism of action. These findings provide insights into combining chemotherapeutic agents with drugs addressing circadian rhythm disorders in CRC treatment strategies. Further research is warranted to elucidate the clinical implications of these observations.

Keywords

• Colorectal Cancer
• Cetuximab
• Agomelatine
• PI3K/AKT/mTOR pathway
• Gene Expression
• Chemotherapy
• Antidepressants
• Combined Therapy.

References

1. Aghamiri, S., Jafarpour, A., Malekshahi, Z. V., Mahmoudi Gomari, M., & Negahdari, B. (2019). Targeting siRNA in colorectal cancer therapy: Nanotechnology comes into view. Journal of Cellular Physiology, 234(9), 14818-14827. https://doi.org/10.1002/jcp.28281
2. Alharbi, K. S., Shaikh, M. A. J., Afzal, O., Altamimi, A. S. A., Almalki, W. H., Alzarea, S. I., Kazmi, I., Al-Abbasi, F. A., Singh, S. K., & Dua, K. (2022). An overview of epithelial growth factor receptor (EGFR) inhibitors in cancer therapy. Chemico-Biological Interactions, 110108. https://doi.org/10.1016/j.cbi.2022.110108
3. Amodio, V., Yaeger, R., Arcella, P., Cancelliere, C., Lamba, S., Lorenzato, A., Arena, S., Montone, M., Mussolin, B., & Bian, Y. (2020). EGFR blockade reverts resistance to KRASG12C inhibition in colorectal cancer. Cancer Discovery, 10(8), 1129-1139. https://doi.org/10.1158/2159-8290.CD-20-0187
4. Ayati, A., Moghimi, S., Salarinejad, S., Safavi, M., Pouramiri, B., & Foroumadi, A. (2020). A review on progression of epidermal growth factor receptor (EGFR) inhibitors as an efficient approach in cancer targeted therapy. Bioorganic Chemistry, 99, 103811. https://doi.org/10.1016/j.bioorg.2020.103811
5. Bu, L.-J., Yu, H.-Q., Fan, L.-L., Li, X.-Q., Wang, F., Liu, J.-T., Zhong, F., Zhang, C.-J., Wei, W., & Wang, H. (2017). Melatoninnatonin, a novel selective ATF-6 inhibitor, induces human hepatoma cell apoptosis through COX-2 downregulation. World Journal of Gastroenterology, 23(6), 986. https://doi.org/ 10.3748/wjg.v23.i6.986
6. Chang, S.-C., and Shen, W. W. (2019). Antidepressant therapy in patients with cancer: a clinical review. Taiwanese Journal of Psychiatry, 33(1), 13-19. https://doi.org/10.4103/TPSY.TPSY_3_19
7. Cho, Y.-S., Yoon, T.-J., Jang, E.-S., Hong, K. S., Lee, S. Y., Kim, O. R., Park, C., Kim, Y.-J., Yi, G.-C., & Chang, K. (2010). Cetuximab-conjugated magneto-fluorescent silica nanoparticles for in vivo colon cancer targeting and imaging. Cancer Letters, 299(1), 63-71. https://doi.org/10.1016/j.canlet.2010.08.004
8. Chok, K. C., Koh, R. Y., Ng, M. G., Ng, P. Y., & Chye, S. M. (2021). Melatonin induces autophagy via reactive oxygen species-mediated endoplasmic reticulum stress pathway in colorectal cancer cells. Molecules, 26(16), 5038. https://doi.org/10.3390/molecules26165038

9. Choudhury, J. D., Kumar, S., Mayank, V., Mehta, J., &Bardalai, D. (2012). A review on apoptosis and its different pathway. International Journal of Biological and Pharmaceutical Research, 3(7), 848-861.
10. Cunningham, D., Humblet, Y., Siena, S., Khayat, D., Bleiberg, H., Santoro, A., Bets, D., Mueser, M., Harstrick, A., & Verslype, C. (2004). Cetuximab monotherapy and cetuximab plus irinotecan in irinotecan-refractory metastatic colorectal cancer. New England Journal of Medicine, 351(4), 337-345. https://doi.org/10.1056/NEJMoa033025
11. Dadsena, S., King, L. E., & García-Sáez, A. J. (2021). Apoptosis regulation at the mitochondria membrane level. Biochimica et Biophysica Acta (BBA)-Biomembranes, 1863(12), 183716. https://doi.org/10.1016/j.bbamem.2021.183716
12. Faivre, S., Kroemer, G., &Raymond, E. (2006). Current development of mTOR inhibitors as anticancer agents. Nature reviews Drug Discovery, 5(8), 671-688. https://doi.org/10.1038/nrd2062
13. Fasolo, A., & Sessa, C. (2008). mTOR inhibitors in the treatment of cancer. Expert Opinion on Investigational Drugs, 17(11), 1717-1734. https://doi.org/10.1517/13543784.17.11.1717
14. Fekry, B., &Eckel-Mahan, K. (2022). The circadian clock and cancer: links between circadian disruption and disease pathology. The Journal of Biochemistry, 171(5), 477-486. https://doi.org/10.1093/jb/mvac017
15. Giordano, G., Remo, A., Porras, A., & Pancione, M. (2019). Immune resistance and EGFR antagonists in colorectal cancer. Cancers, 11(8), 1089. https://doi.org/10.3390/cancers11081089
16. Han, H., Li, Y., Qin, W., Wang, L., Yin, H., Su, B., & Yuan, X. (2022). miR-199b-3p contributes to acquired resistance to cetuximab in colorectal cancer by targeting CRIM1 via Wnt/β-catenin signaling. Cancer Cell International, 22(1), 42. https://doi.org/10.1186/s12935-022-02460-x
17. Hanck-Silva, G., Fatori Trevizan, L. N., Petrilli, R., de Lima, F. T., Eloy, J. O., & Chorilli, M. (2020). A Critical review of properties and analytical/bioanalytical methods for characterization of cetuximab. Critical Reviews in Analytical Chemistry, 50(2), 125-135. https://doi.org/10.1080/10408347.2019.1581984
18. Hossain, M. S., Karuniawati, H., Jairoun, A. A., Urbi, Z., Ooi, D. J., John, A., Lim, Y. C., Kibria, K. K., Mohiuddin, A., & Ming, L. C. (2022). Colorectal cancer: a review of carcinogenesis, global epidemiology, current challenges, risk factors, preventive and treatment strategies. Cancers, 14(7), 1732. https://doi.org/10.3390/cancers14071732
19. Kılıç, N., &Erbaş, O. (2021). Antidepressant Drugs, Biological Clocks, and Cancer: Is There a Relation? Journal of Experimental and Basic Medical Sciences, 2(3), https://doi.org/298-301. 10.5606/jebms.2021.75670
20. Li, R., Liang, M., Liang, X., Yang, L., Su, M., & Lai, K. P. (2020). Chemotherapeutic effectiveness of combining cetuximab for metastatic colorectal cancer treatment: A system review and meta-analysis. Frontiers in Oncology, 10, 868. https://doi.org/10.3389/fonc.2020.00868
21. Li, X., Zhao, L., Chen, C., Nie, J., & Jiao, B. (2022). Can EGFR be a therapeutic target in breast cancer? Biochimica et Biophysica Acta (BBA)-Reviews on Cancer, 188789. https://doi.org/10.1016/j.bbcan.2022.188789
22. Livak, K. J., & Schmittgen, T. D. (2001). Analysis of relative gene expression data using real-time quantitative PCR and the 2− ΔΔCT method. Methods, 25(4), 402-408. https://doi.org/10.1006/meth.2001.1262
23. Ma, Q., Reiter, R. J., & Chen, Y. (2020). Role of melatoninnatonin in controlling angiogenesis under physiological and pathological conditions. Angiogenesis, 23, 91-104. https://doi.org/10.1007/s10456-019-09689-7
24. Miricescu, D., Totan, A., Stanescu-Spinu, I.-I., Badoiu, S. C., Stefani, C., & Greabu, M. (2020). PI3K/AKT/mTOR signaling pathway in breast cancer: from molecular landscape to clinical aspects. International Journal of Molecular Sciences, 22(1), 173. https://doi.org/10.3390/ijms22010173
25. Moreno-SanJuan, S., Puentes-Pardo, J. D., Casado, J., Escudero-Feliu, J., Khaldy, H., Arnedo, J., Carazo, Á., & León, J. (2023). Agomelatine, a melatonin-derived drug, as a new strategy for the treatment of colorectal cancer. Antioxidants, 12(4), 926. https://doi.org/10.3390/antiox12040926
26. Naser, A. Y., Hameed, A. N., Mustafa, N., Alwafi, H., Dahmash, E. Z., Alyami, H. S., & Khalil, H. (2021). Depression and anxiety in patients with cancer: a cross-sectional study. Frontiers in Psychology, 12, 1067. https://doi.org/10.3389/fpsyg.2021.585534
27. Ogrodnik, M. (2021). Cellular aging beyond cellular senescence: Markers of senescence prior to cell cycle arrest in vitro and in vivo. Aging Cell, 20(4), e13338. https://doi.org/10.1111/acel.13338
28. Revathidevi, S., & Munirajan, A. K. (2019). Akt in cancer: Mediator and more. Seminars in cancer biolog, 59:80-91. https://doi.org/10.1016/j.semcancer.2019.06.002
29. Sartore-Bianchi, A., Trusolino, L., Martino, C., Bencardino, K., Lonardi, S., Bergamo, F., Zagonel, V., Leone, F., Depetris, I., & Martinelli, E. (2016). Dual-targeted therapy with trastuzumab and lapatinib in treatment-refractory, KRAS codon 12/13 wild-type, HER2-positive metastatic colorectal cancer (HERACLES): a proof-of-concept, multicentre, open-label, phase 2 trial. The Lancet Oncology, 17(6), 738-746. https://doi.org/10.1016/S1470-2045(16)00150-9
30. Tran, Q. H., Hoang, D. H., Song, M., Choe, W., Kang, I., Kim, S. S., & Ha, J. (2021). Melatoninnatonin and doxorubicin synergistically enhance apoptosis via autophagy-dependent reduction of AMPKα1 transcription in human breast cancer cells. Experimental and Molecular Medicine, 53(9), 1413-1422. https://doi.org/10.1038/s12276-021-00675-y
31. Wathoni, N., Nguyen, A. N., Rusdin, A., Umar, A. K., Mohammed, A. F. A., Motoyama, K., Joni, I. M., & Muchtaridi, M. (2020). Enteric-coated strategies in colorectal cancer nanoparticle drug delivery system. Drug Design, Development and Therapy, 4387-4405. https://doi.org/10.2147/DDDT.S273612
32. Xi, Y., & Xu, P. (2021). Global colorectal cancer burden in 2020 and projections to 2040. Translational Oncology, 14(10), 101174. https://doi.org/10.1016/j.tranon.2021.101174
33. Zhao, Y., Wang, C., & Goel, A. (2022). A combined treatment with melatoninnatonin and andrographis promotes autophagy and anticancer activity in colorectal cancer. Carcinogenesis, 43(3), 217-230. https://doi.org/10.1093/carcin/bgac008