Effects of r-Klotho on MMP/TIMP Balance and TNF-α in Colon Cancer and Normal Colon Epithelial Cells

Objective: To evaluate the effects of recombinant klotho protein (r-klotho) on human colon adenocarcinoma cells (Caco-2) and normal colon epithelial cells (CCD 841 CoN), with a focus on extracellular matrix (ECM) remodeling and inflammation-related gene expression. Materials and Methods: Caco-2 and CCD 841 CoN cells were cultured in four experimental groups: CCD 841 CoN; Caco-2; r-klotho-treated CCD 841 CoN; and r-klotho-treated Caco-2. r-klotho was administered at 0.3 μg/mL, selected from a previous dose–response study as the concentration yielding the lowest relative viability in Caco-2 cells within the tested range, for 24 and 48 hours. Expression levels of MMP-1, MMP-3, MMP-13, TIMP-1, TIMP-3, and TNF-α were analysed using quantitative real-time PCR (qRT-PCR) and evaluated by the ΔΔCt method with GAPDH as the reference gene. Results: In Caco-2 cells, r-klotho significantly reduced MMP-1 and TIMP-1 expression (p