Synergistic Induction of Colorectal Cancer Cell Proliferation by ox-LDL and TNF-α

Colorectal cancer is the third most commonly diagnosed malignancy and the second leading cause of cancer-related death 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 elevated levels of tumor necrosis factor alpha (TNF-α) and oxidized low-density lipoprotein (ox-LDL).The objective of this study was to analyze the synergistic effect of ox-LDL and TNF-α on ROS production and cell proliferation via WNT/β-catenin and PI3K/AKT in colorectal cancer. To this end, the CRC cells, COLO320 and SW620, were treated with various concentrations of ox-LDL, TNF-α, and their combinations. The pro-liferative effect induced by their synergism was assessed using the IncuCyte® Real-Time Assay, as well as ROS generation was measured with the 2′,7′-dichlorodihydrofluorescein diacetate (H2DCFDA) probe, and cell viability was evaluated by MTT under conditions of pathway inhibition. Co-treatment with ox-LDL and TNF-α significantly increased proliferation in COLO320 cells, accompanied by a marked rise in ROS generation in both cell lines. Inhibition of 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.