Abstract
Oral squamous cell carcinoma (OSCC) is the most common type of head and neck squamous cell carcinoma (HNSCC), and the effect of zymosan (ZYM), a component of the yeast cell wall, on oral cancer remains unclear. The CCK-8 proliferation assay was performed to evaluate the effect of ZYM on the proliferation of the OSCC cell lines WSU-HN4, WSU-HN6 and CAL27, and the potential mechanism was explored by quantitative real-time PCR, immunofluorescence assay and western blot. A cell adhesion assay was conducted to determine the adhesion of Candida albicans to OSCC cells, and the expression of related genes, including TLR2, MyD88, NLRP3, ASC, Caspase-1 and IL-1β, and proteins, including TLR2, MyD88, NF-κB p65, p-NF-κB p65 and E-cadherin was determined. Additionally, the pro-inflammatory cytokines including IL-6, IL-8, TNF-α and IL-1β produced by OSCC cells were detected using a chemiluminescence immunoassay (CLIA). In the current study, the CCK-8 assay showed that ZYM promoted the proliferation of WSU-HN4, WSU-HN6 and CAL27 cells via the TLR2/MyD88 pathway. The cell adhesion assay showed that the number of C. albicans cells per field significantly increased in ZYM-treated OSCC cells compared to controls. When treated with ZYM, OSCC cells secreted significantly more pro-inflammatory cytokine IL-1β, which could enhance inflammation in oral cancer microenvironment. In conclusion, ZYM from the fungal cell wall promotes the proliferation, C. albicans adhesion and IL-1β production in OSCC, as demonstrated by in vitro experiments.
Highlights
Oral squamous cell carcinoma (OSCC) is the most common type of head and neck squamous cell carcinoma (HNSCC), which is the sixth leading cancer type worldwide [1]
Zymosan from the fungal cell wall promotes the proliferation of OSCC To investigate the effect of ZYM on oral cancer cells, we cultured OSCC cell lines (WSU-HN4, WSU-HN6 and CAL27) with zymosan at the concentrations of 10 μg/ml and 100 μg/ml for 12 h, 24 h or 48 h
The TLR2/myeloid differentiation primary response protein 88 (MyD88) pathway is involved in the interaction between OSCC and zymosan To define the receptors of zymosan in OSCC cells, the mRNA expression of receptors including TLR2, TLR6, Dectin-1 and CR3 was detected by quantitative real-time PCR (RT-PCR)
Summary
Oral squamous cell carcinoma (OSCC) is the most common type of head and neck squamous cell carcinoma (HNSCC), which is the sixth leading cancer type worldwide [1]. It has been observed that oral carriage of C. albicans is higher in patients with OSCC or leukoplakia than in those without oral pathology [11, 12], but the association between C. albicans and oral cancer remains ambiguous. Several reports have proposed different potential pathways by which C. albicans initiates or promotes carcinogenesis, such as inducing inflammation [13, 14]. The adhesion of microorganisms to host cells has been reported to be the first step for bacteria or fungi to influence host cells [8]. The adhesion of C. albicans to OSCC cells remains unclear
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