Chemotherapy drugs and inflammatory cytokines enhance matrix metalloproteinases expression by oral mucosa cells

Abstract

Objective: Oral mucositis (OM), the most common side effect of cancer therapy, is associated with pro-inflammatory cytokines and matrix metalloproteinases (MMPs) increased expression. Although there are approaches for OM management, none is infallible, thus, elucidation of molecular events related to OM etiopathogenesis may improve current therapeutic strategies. This study assessed the influence of pro-inflammatory cytokines and chemotherapy drugs on MMPs expression and synthesis by oral mucosa cells. Design: Human gingival fibroblasts (HGF) were exposed to different concentrations of methotrexate (MTX) and 5-fluorouracil (5-FU); subsequentially, cell viability, nitric oxide and interleukin(IL)-6 production were evaluated to select the concentration of these drugs that could stimulate inflammatory phenotype without cytotoxic effects. Then, HGF and primary gingival keratinocytes (PGK) were subjected to different stimuli: culture medium (negative control), tumor necrosis factor-alpha (TNF-α – positive control), IL-6, IL-8, MTX, and 5-FU for 3, 6, 12, and 24 h. Next, gene expression and synthesis of MMP-2 and MMP-9 by HGF and MMP-3 by PGK were assessed. Results: At 6 h, MMP-2 synthesis increased 60 % after exposure to TNF-α and MTX, 40 % after IL-6, and 15 % after IL-8. At 12 h, MMP-9 synthesis increased 15 % after exposure to TNF-α, while MMP-3 synthesis increased 30 % after TNF-α, and 10 % after IL-8. TNF-α-treated groups presented increased gene expression of all MMPs evaluated. IL-8 and 5-FU increased MMP-2 and MMP-3 expression, while IL-6 and MTX augmented MMP-2 expression. Conclusions: The chemotherapy drugs and cytokines investigated up-regulated MMPs expression by oral mucosa cells, which may lead to OM establishment and severity.