Synergistic induction of colorectal cancer cell proliferation by ox-LDL and TNF-α.

Authors

  • María Palma-Vejares Biotechnology and Biopharmaceutical Laboratory, Departamento de Fisiopatología, Facultad de Ciencias Biológicas, Universidad de Concepción, Víctor Lamas 1290, P.O. Box 160-C, Concepción 4030000, Chile https://orcid.org/0009-0000-2481-826X
  • Elizabeth Santana Biotechnology and Biopharmaceutical Laboratory, Departamento de Fisiopatología, Facultad de Ciencias Biológicas, Universidad de Concepción, Víctor Lamas 1290, P.O. Box 160-C, Concepción 4030000, Chile https://orcid.org/0009-0002-6372-9790
  • Carla Villavicencio Biotechnology and Biopharmaceutical Laboratory, Departamento de Fisiopatología, Facultad de Ciencias Biológicas, Universidad de Concepción, Víctor Lamas 1290, P.O. Box 160-C, Concepción 4030000, Chile.
  • Angela Hidalgo-Gajardo Biotechnology and Biopharmaceutical Laboratory, Departamento de Fisiopatología, Facultad de Ciencias Biológicas, Universidad de Concepción, Víctor Lamas 1290, P.O. Box 160-C, Concepción 4030000, Chile. https://orcid.org/0000-0003-0600-5243
  • Jorge R. Toledo Biotechnology and Biopharmaceutical Laboratory, Departamento de Fisiopatología, Facultad de Ciencias Biológicas, Universidad de Concepción, Víctor Lamas 1290, P.O. Box 160-C, Concepción 4030000, Chile https://orcid.org/0000-0002-9879-7710

DOI:

https://doi.org/10.70099/BJ/2025.02.03.14

Keywords:

Colorectal cancer, proliferation, ROS, TNF-α, ox-LDL

Abstract

Colorectal cancer (CRC) is the third most commonly diagnosed malignancy and the second leading cause of cancer-related deaths worldwide. Its incidence continues to rise, particularly in association with modifiable risk factors such as obesity, which is closely linked to chronic inflammation and metabolic disturbances, including dyslipidemia. These conditions contribute to the formation of a pro-inflammatory tumor microenvironment, characterized by high levels of TNF-α and ox-LDL. This study aimed to analyze the synergistic effects of ox-LDL and TNF-α on ROS production and cell proliferation via the WNT/β-catenin and PI3K/AKT pathways in CRC cells. COLO320 and SW620 cells were treated with various concentrations of ox-LDL, TNF-α, and their combinations. The proliferation induced was assessed using the IncuCyte® Real-Time Assay. ROS generation was measured using the 2′,7′-dichlorodihydrofluorescein diacetate (H2DCFDA) probe. Cell viability was evaluated using the MTT assay under conditions of pathway inhibition. Co-treatment with ox-LDL and TNF-α significantly increased proliferation in COLO320 cells, and was accompanied by a marked increase in ROS generation in both cell lines. Inhibiting the WNT/β-catenin and PI3K/AKT pathways revealed differential responses, suggesting a heterogeneous activation pattern dependent on the molecular context. To our knowledge, this is the first study to demonstrate the synergistic effect of ox-LDL and TNF-α in colorectal cancer cell models. These findings highlight the importance of considering both the molecular and redox context of the tumor microenvironment when designing personalized therapeutic strategies.

References

1. Brown, J.S., Amend, S.R., Austin, R.H., Gatenby, R.A., Hammarlund, E.U., and Pienta, K.J., Updating the Definition of Cancer. Mol Cancer Res, 2023. 21(11): p. 1142-1147.

2. Organization, W.H. Cancer. 2022; Available from: https://www.who.int/es/news-room/fact-sheets/detail/cancer.

3. Bray, F., Laversanne, M., Sung, H., Ferlay, J., Siegel, R.L., Soerjomataram, I., and Jemal, A., Global cancer statistics 2022: GLOBOCAN estimates of incidence and mortality worldwide for 36 cancers in 185 countries. CA Cancer J Clin, 2024. 74(3): p. 229-263.

4. Crockett, S.D. and Nagtegaal, I.D., Terminology, Molecular Features, Epidemiology, and Management of Serrated Colorectal Neoplasia. Gastroenterology, 2019. 157(4): p. 949-966 e944.

5. Sedlak, J.C., Yilmaz, O.H., and Roper, J., Metabolism and Colorectal Cancer. Annu Rev Pathol, 2023. 18: p. 467-492.

6. Goosenberg, E., Kaur, A., and Babiker, H.M., A Review of Hereditary Colorectal Cancers, in StatPearls. 2025: Treasure Island (FL).

7. Keum, N. and Giovannucci, E., Global burden of colorectal cancer: emerging trends, risk factors and prevention strategies. Nat Rev Gastroenterol Hepatol, 2019. 16(12): p. 713-732.

8. González-Horta E., V.Y., Burgueño J., Abreu M. and Toledo J., P-305 Oxidized-LDL promotes colorectal cancer progression and growth of human colonoides. Annals of Oncology, 2022. 33.

9. Gonzalez-Horta, E.E., Burgueno, J.F., Leiva, M.J., Villavicencio, C., Kawaguchi, F.I., Hazime, H., Reyes, F., Manrique-Suarez, V., Parra, N.C., Abreu, M.T., and Toledo, J.R., Oxidized Low-Density Lipoprotein Induces Reactive Oxygen Species-Dependent Proliferation of Intestinal Epithelial Cells. Pharmaceuticals (Basel), 2024. 17(11).

10. Catalano, T., Selvaggi, F., Cotellese, R., and Aceto, G.M., The Role of Reactive Oxygen Species in Colorectal Cancer Initiation and Progression: Perspectives on Theranostic Approaches. Cancers (Basel), 2025. 17(5).

11. Zhao, T.L., Qi, Y., Wang, Y.F., Wang, Y., Liang, H., and Pu, Y.B., 5-methoxytryptophan induced apoptosis and PI3K/Akt/FoxO3a phosphorylation in colorectal cancer. World J Gastroenterol, 2023. 29(47): p. 6148-6160.

12. Akhiani, A.A. and Martner, A., Role of Phosphoinositide 3-Kinase in Regulation of NOX-Derived Reactive Oxygen Species in Cancer. Antioxidants (Basel), 2022. 12(1).

13. Disoma, C., Zhou, Y., Li, S., Peng, J., and Xia, Z., Wnt/beta-catenin signaling in colorectal cancer: Is therapeutic targeting even possible? Biochimie, 2022. 195: p. 39-53.

14. Staehlke, S., Haack, F., Waldner, A.C., Koczan, D., Moerke, C., Mueller, P., Uhrmacher, A.M., and Nebe, J.B., ROS Dependent Wnt/beta-Catenin Pathway and Its Regulation on Defined Micro-Pillars-A Combined In Vitro and In Silico Study. Cells, 2020. 9(8).

15. Basak, D., Uddin, M.N., and Hancock, J., The Role of Oxidative Stress and Its Counteractive Utility in Colorectal Cancer (CRC). Cancers (Basel), 2020. 12(11).

16. Chen, X., Andresen, B.T., Hill, M., Zhang, J., Booth, F., and Zhang, C., Role of Reactive Oxygen Species in Tumor Necrosis Factor-alpha Induced Endothelial Dysfunction. Curr Hypertens Rev, 2008. 4(4): p. 245-255.

17. Cominacini, L., Pasini, A.F., Garbin, U., Davoli, A., Tosetti, M.L., Campagnola, M., Rigoni, A., Pastorino, A.M., Lo Cascio, V., and Sawamura, T., Oxidized low density lipoprotein (ox-LDL) binding to ox-LDL receptor-1 in endothelial cells induces the activation of NF-kappaB through an increased production of intracellular reactive oxygen species. J Biol Chem, 2000. 275(17): p. 12633-12638.

18. Eswar, S., Rajagopalan, B., Ete, K., and Nageswara Rao Gattem, S., Serum Tumor Necrosis Factor Alpha (TNF-alpha) Levels in Obese and Overweight Adults: Correlations With Metabolic Syndrome and Inflammatory Markers. Cureus, 2024. 16(7): p. e64619.

19. Freitas, M.C.P., Fernandez, D.G.E., Cohen, D., Figueiredo-Neto, A.M., Maranhao, R.C., and Damasceno, N.R.T., Oxidized and electronegative low-density lipoprotein as potential biomarkers of cardiovascular risk in obese adolescents. Clinics (Sao Paulo), 2018. 73: p. e189.

20. Gupta, S., Screening for Colorectal Cancer. Hematol Oncol Clin North Am, 2022. 36(3): p. 393-414.

21. Li, H., Li, X.X., Ma, Q., and Cui, J., The variability of oxLDL-induced cytotoxicity on different types of cell lines. Cell Biochem Biophys, 2013. 67(2): p. 635-644.

22. Gonzalez-Chavarria, I., Cerro, R.P., Parra, N.P., Sandoval, F.A., Zuniga, F.A., Omazabal, V.A., Lamperti, L.I., Jimenez, S.P., Fernandez, E.A., Gutierrez, N.A., Rodriguez, F.S., Onate, S.A., Sanchez, O., Vera, J.C., and Toledo, J.R., Lectin-like oxidized LDL receptor-1 is an enhancer of tumor angiogenesis in human prostate cancer cells. PLoS One, 2014. 9(8): p. e106219.

23. Cai, X., Cao, C., Li, J., Chen, F., Zhang, S., Liu, B., Zhang, W., Zhang, X., and Ye, L., Inflammatory factor TNF-alpha promotes the growth of breast cancer via the positive feedback loop of TNFR1/NF-kappaB (and/or p38)/p-STAT3/HBXIP/TNFR1. Oncotarget, 2017. 8(35): p. 58338-58352.

24. Chou, C.C., Wang, C.P., Chen, J.H., and Lin, H.H., Anti-Atherosclerotic Effect of Hibiscus Leaf Polyphenols against Tumor Necrosis Factor-alpha-Induced Abnormal Vascular Smooth Muscle Cell Migration and Proliferation. Antioxidants (Basel), 2019. 8(12).

25. Zhao, P. and Zhang, Z., TNF-alpha promotes colon cancer cell migration and invasion by upregulating TROP-2. Oncol Lett, 2018. 15(3): p. 3820-3827.

26. Zhang, X., Ye, X., and Jin, H., Oxidized Low-Density Lipoprotein as a Potential Target for Enhancing Immune Checkpoint Inhibitor Therapy in Microsatellite-Stable Colorectal Cancer. Antioxidants (Basel), 2025. 14(6).

27. Echizen, K., Horiuchi, K., Aoki, Y., Yamada, Y., Minamoto, T., Oshima, H., and Oshima, M., NF-kappaB-induced NOX1 activation promotes gastric tumorigenesis through the expansion of SOX2-positive epithelial cells. Oncogene, 2019. 38(22): p. 4250-4263.

28. Kuwano, Y., Tominaga, K., Kawahara, T., Sasaki, H., Takeo, K., Nishida, K., Masuda, K., Kawai, T., Teshima-Kondo, S., and Rokutan, K., Tumor necrosis factor alpha activates transcription of the NADPH oxidase organizer 1 (NOXO1) gene and upregulates superoxide production in colon epithelial cells. Free Radic Biol Med, 2008. 45(12): p. 1642-1652.

29. Konate, M.M., Antony, S., and Doroshow, J.H., Inhibiting the Activity of NADPH Oxidase in Cancer. Antioxid Redox Signal, 2020. 33(6): p. 435-454.

30. Gianni, D., Taulet, N., Zhang, H., DerMardirossian, C., Kister, J., Martinez, L., Roush, W.R., Brown, S.J., Bokoch, G.M., and Rosen, H., A novel and specific NADPH oxidase-1 (Nox1) small-molecule inhibitor blocks the formation of functional invadopodia in human colon cancer cells. ACS Chem Biol, 2010. 5(10): p. 981-993.

31. Chen, G.T., Tifrea, D.F., Murad, R., Habowski, A.N., Lyou, Y., Duong, M.R., Hosohama, L., Mortazavi, A., Edwards, R.A., and Waterman, M.L., Disruption of beta-Catenin-Dependent Wnt Signaling in Colon Cancer Cells Remodels the Microenvironment to Promote Tumor Invasion. Mol Cancer Res, 2022. 20(3): p. 468-484.

32. Castel, P., Ellis, H., Bago, R., Toska, E., Razavi, P., Carmona, F.J., Kannan, S., Verma, C.S., Dickler, M., Chandarlapaty, S., Brogi, E., Alessi, D.R., Baselga, J., and Scaltriti, M., PDK1-SGK1 Signaling Sustains AKT-Independent mTORC1 Activation and Confers Resistance to PI3Kalpha Inhibition. Cancer Cell, 2016. 30(2): p. 229-242.

33. Clarke, P.A., Roe, T., Swabey, K., Hobbs, S.M., McAndrew, C., Tomlin, K., Westwood, I., Burke, R., van Montfort, R., and Workman, P., Dissecting mechanisms of resistance to targeted drug combination therapy in human colorectal cancer. Oncogene, 2019. 38(25): p. 5076-5090.

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Published

2025-09-15

How to Cite

Palma-Vejares, M., Santana, E., Villavicencio, C., Hidalgo-Gajardo, A., & Toledo, J. R. (2025). Synergistic induction of colorectal cancer cell proliferation by ox-LDL and TNF-α. BioNatura Journal: Ibero-American Journal of Biotechnology and Life Sciences, 2(3), 15. https://doi.org/10.70099/BJ/2025.02.03.14

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