estuscience - life Eskişehir Teknik Üniversitesi Bilim ve Teknoloji Dergisi - C Yaşam Bilimleri Ve Biyoteknoloji 2667-4203 Eskişehir Teknik Üniversitesi 10.18036/estubtdc.1798104 Cell Metabolism Hücre Metabolizması THE LIPOXYGENASE INHIBITOR ML351 AFFECTS CELL VIABILITY BY REGULATING AUTOPHAGY AND SEMAPHORIN SIGNALING PATHWAYS IN A549 CELLS THE LIPOXYGENASE INHIBITOR ML351 AFFECTS CELL VIABILITY BY REGULATING AUTOPHAGY AND SEMAPHORIN SIGNALING PATHWAYS IN A549 CELLS https://orcid.org/0000-0002-2878-5052 Meriç Neslihan Kütahya Sağlık Bilimleri Üniversitesi https://orcid.org/0000-0003-2134-4067 Kar Ezgi KUTAHYA HEALTH SCIENCES UNIVERSITY https://orcid.org/0000-0002-0993-6118 Hacıoğlu Ceyhan DUZCE UNIVERSITY https://orcid.org/0000-0001-8356-9806 Kar Fatih KUTAHYA HEALTH SCIENCES UNIVERSITY 01 27 2026 15 1 74 82 10 06 2025 10 27 2025 Copyright © 2010, Eskişehir Teknik Üniversitesi Bilim ve Teknoloji Dergisi - C Yaşam Bilimleri Ve Biyoteknoloji 2010 Eskişehir Teknik Üniversitesi Bilim ve Teknoloji Dergisi - C Yaşam Bilimleri Ve Biyoteknoloji

Lung cancer is one of the leading causes of cancer-related deaths worldwide, and the inadequacy of current treatment strategies necessitates the investigation of new molecular targets. In this study, the effects of the selective 15-lipoxygenase inhibitor ML351 on cell viability, autophagy, and the Semaphorin-3E (SEMA3E) signaling pathway in A549 lung adenocarcinoma cells were examined.Cell viability was evaluated using the MTT assay, and protein levels of autophagy markers (Beclin-1, LC3, and p62) and SEMA3E were measured using the ELISA method. The findings revealed that ML351 exhibited a dose-dependent cytotoxic effect. The IC₅₀ value was calculated as 92 µM, cell viability fell below 80% in the 20–80 µM range, and was almost completely eliminated at 160 µM (***p

Lung cancer is one of the leading causes of cancer-related deaths worldwide, and the inadequacy of current treatment strategies necessitates the investigation of new molecular targets. In this study, the effects of the selective 15-lipoxygenase inhibitor ML351 on cell viability, autophagy, and the Semaphorin-3E (SEMA3E) signaling pathway in A549 lung adenocarcinoma cells were examined.Cell viability was evaluated using the MTT assay, and protein levels of autophagy markers (Beclin-1, LC3, and p62) and SEMA3E were measured using the ELISA method. The findings revealed that ML351 exhibited a dose-dependent cytotoxic effect. The IC₅₀ value was calculated as 92 µM, cell viability fell below 80% in the 20–80 µM range, and was almost completely eliminated at 160 µM (***p

 ML351 Autophagy SEMA3E A549 ML351 Autophagy SEMA3E A549 [1] Bray F, Ferlay J, Soerjomataram I, Siegel RL, Torre LA, Jemal A. Global cancer statistics 2018: GLOBOCAN estimates of incidence and mortality worldwide for 36 cancers in 185 countries. CA Cancer J Clin. 2018;68(6):394-424. doi:10.3322/caac.21492 [2] Siegel RL, Miller KD, Jemal A. Cancer statistics, 2020. CA Cancer J Clin. 2020;70(1):7-30. doi:10.3322/caac.21590 [3] García-Campelo R, Bernabé R, Cobo M, et al. SEOM clinical guidelines for the treatment of non-small cell lung cancer (NSCLC) 2015. Clin Transl Oncol. 2015;17(12):1020-1029. doi:10.1007/s12094-015-1455-z [4] Mizushima N, Komatsu M. Autophagy: renovation of cells and tissues. Cell. 2011;147(4):728-741. doi:10.1016/j.cell.2011.10.026 [5] Jalali P, Shahmoradi A, Samii A, et al. The role of autophagy in cancer: from molecular mechanism to therapeutic window. Front Immunol. 2025;16. doi:10.3389/fimmu.2025.1528230 [6] Santana-Codina N, Mancias JD, Kimmelman AC. The Role of Autophagy in Cancer. Annual Review of Cancer Biology. 2017;1(Volume 1, 2017):19-39. doi:10.1146/annurev-cancerbio-041816-122338 [7] Mowers EE, Sharifi MN, Macleod KF. Autophagy in cancer metastasis. Oncogene. 2017;36(12):1619-1630. doi:10.1038/onc.2016.333 [8] Hassan M, Watari H, AbuAlmaaty A, Ohba Y, Sakuragi N. Apoptosis and molecular targeting therapy in cancer. Biomed Res Int. 2014;2014:150845. doi:10.1155/2014/150845 [9] Molero-Valenzuela A, Fontova P, Alonso-Carrillo D, et al. A Novel Late-Stage Autophagy Inhibitor That Efficiently Targets Lysosomes Inducing Potent Cytotoxic and Sensitizing Effects in Lung Cancer. Cancers (Basel). 2022;14(14):3387. doi:10.3390/cancers14143387 [10] Schneider C, Pozzi A. Cyclooxygenases and lipoxygenases in cancer. Cancer Metastasis Rev. 2011;30(3-4):277-294. doi:10.1007/s10555-011-9310-3 [11] Melstrom LG, Bentrem DJ, Salabat MR, et al. Overexpression of 5-lipoxygenase in colon polyps and cancer and the effect of 5-LOX inhibitors in vitro and in a murine model. Clin Cancer Res. 2008;14(20):6525-6530. doi:10.1158/1078-0432.CCR-07-4631 [12] Neufeld G, Mumblat Y, Smolkin T, et al. The role of the semaphorins in cancer. Cell Adhesion & Migration. 2016;10(6):652-674. doi:10.1080/19336918.2016.1197478 [13] Valentini E, Di Martile M, Del Bufalo D, D’Aguanno S. SEMAPHORINS and their receptors: focus on the crosstalk between melanoma and hypoxia. Journal of Experimental & Clinical Cancer Research. 2021;40(1):131. doi:10.1186/s13046-021-01929-3 [14] Neufeld G, Kessler O. The semaphorins: versatile regulators of tumour progression and tumour angiogenesis. Nat Rev Cancer. 2008;8(8):632-645. doi:10.1038/nrc2404 [15] Nakayama H, Murakami A, Nishida-Fukuda H, et al. Semaphorin 3F inhibits breast cancer metastasis by regulating the Akt-mTOR and TGFβ signaling pathways via neuropilin-2. Sci Rep. 2025;15(1):7394. doi:10.1038/s41598-025-91559-y [16] Jiang J, Zhang F, Wan Y, et al. Semaphorins as Potential Immune Therapeutic Targets for Cancer. Front Oncol. 2022;12:793805. doi:10.3389/fonc.2022.793805 [17] Yang J, Zeng Z, Qiao L, et al. Semaphorin 4C Promotes Macrophage Recruitment and Angiogenesis in Breast Cancer. Mol Cancer Res. 2019;17(10):2015-2028. doi:10.1158/1541-7786.MCR-18-0933 [18] Rai G, Joshi N, Perry S, et al. Discovery of ML351, a Potent and Selective Inhibitor of Human 15-Lipoxygenase-1. In: Probe Reports from the NIH Molecular Libraries Program. National Center for Biotechnology Information (US); 2010. Accessed October 3, 2025. http://www.ncbi.nlm.nih.gov/books/NBK190602/ [19] Casazza A, Finisguerra V, Capparuccia L, et al. Sema3E-Plexin D1 signaling drives human cancer cell invasiveness and metastatic spreading in mice. J Clin Invest. 2010;120(8):2684-2698. doi:10.1172/JCI42118 [20] Luchino J, Hocine M, Amoureux MC, et al. Semaphorin 3E suppresses tumor cell death triggered by the plexin D1 dependence receptor in metastatic breast cancers. Cancer Cell. 2013;24(5):673-685. doi:10.1016/j.ccr.2013.09.010 [21] Bhutia SK, Mukhopadhyay S, Sinha N, et al. Autophagy: Cancer’s Friend or Foe? Adv Cancer Res. 2013;118:61-95. doi:10.1016/B978-0-12-407173-5.00003-0 [22] Mulcahy Levy JM, Thorburn A. Autophagy in cancer: moving from understanding mechanism to improving therapy responses in patients. Cell Death Differ. 2020;27(3):843-857. doi:10.1038/s41418-019-0474-7 [23] Dobrian AD, Morris MA, Taylor-Fishwick DA, et al. Role of the 12-lipoxygenase pathway in diabetes pathogenesis and complications. Pharmacology & Therapeutics. 2019;195:100-110. doi:10.1016/j.pharmthera.2018.10.010 [24] Snodgrass RG, Zezina E, Namgaladze D, et al. A Novel Function for 15-Lipoxygenases in Cholesterol Homeostasis and CCL17 Production in Human Macrophages. Front Immunol. 2018;9. doi:10.3389/fimmu.2018.01906