Role of Oxidative Stress-Dependent C/EBPβ Expression on CAF Transformation Inducing HCT116 Colorectal Cancer Cell Progression; Migration and Invasion

Abstract

OBJECTIVE: To investigate oxidative stress-related CAF transformation through C/EBPβ, which affects CRC progression and may have a potential implication for CRC treatment. METHODS: The conditioned media (CM) from HCT116, CRC cells, was used to activate CCD-18Co, colon fibroblasts, then the ability of activated FBs to induce HCT116 growth and progression was assessed using MTT assay, transwell migration, and matrix invasion assay. Alteration of the cytokine profile and oxidative stress of the activated FBs were studied with cytokine arrays and DCFH-DA assay, respectively. The protein expressions of the CAF markers (α-SMA and FAP) and C/EBPβ were investigated with immunofluorescence and western blotting. RESULT: It was found that CM from HCT116 cells induced oxidative stress, change of cytokine profile, CAF markers, and the C/EBPβ expression of activated FBs. Furthermore, when the oxidative stress of the activated FBs was suppressed, FAP and C/EBPβ expression were downregulated, correlating with the disabling of their capability to support the cancer progression. The C/EBPβ and prognosis for CRC patients were accessed using the GEPIA dataset, in which high C/EBPβ expression was associated with a poor prognosis. CONCLUSION: These findings suggest that C/EBPβ expression has a role in CAF transformation in an oxidative stress-related manner and might be used as a target to improve aggressive CRC treatment outcomes.