Promoter hypermethylation may contribute to E‑cadherin repression in the human salivary carcinoma ex pleomorphic adenoma
- Liang Xia
- Yuhua Hu
- Ting Gu
- Lizhen Wang
- Zhen Tian
Published online on: November 22, 2017
The role of promoter methylation in the inactivation of E‑cadherin (CDH1) in salivary carcinoma ex pleomorphic adenoma (CXPA) is unknown. The objective of this study was to determine the role and potential clinical implications of CDH1 promoter methylation in salivary CXPA. The CDH1 promoter methylation status was determined by bisulfite sequencing PCR in 37 primary CXPA tissues and 2 CXPA cell lines. E‑cadherin expression levels were determined by immunohistochemical analysis of each tumor. E‑cadherin protein levels and CDH1 mRNA levels were examined by immunoblotting and quantitative real-time PCR, respectively, in 2 CXPA cell lines. Cells were treated with 5‑Aza-dC or TGF‑β1 to test the influence of promoter methylation on CDH1 mRNA and protein expression. Associations between CDH1 molecular alterations and patients' clinicopathologic characteristics and prognosis were statistically evaluated. CDH1 promoter hypermethylation was detected in 21 of 37 tumors (56.76%). Of these 37 tumors, 13 tumors (35.14%) showed low E‑cadherin expression. Tumors that had CDH1 promoter methylation had a histological tendency toward luminal differentiation (P=0.004), high tumor grade (P=0.005), high T stage (P=0.024) and high TNM stage (P=0.038) compared with tumors that did not. The two CXPA cell lines exhibited an inverse relationship between CDH1 promoter methylation status and CDH1 mRNA and protein expression. Treatment of the hypermethylated cell line with 5‑Aza-dC restored CDH1 mRNA and E‑cadherin protein expression. The induction of hypermethylation by TGF‑β1 resulted in the repression of CDH1 mRNA and protein expression. CDH1 is commonly silenced in CXPA through promoter methylation. CDH1 methylation is closely related to tumor cell differentiation, histological grade, lymph node metastasis and advanced TNM stage, indicating that CDH1 methylation may play a role in the initiation and progression of CXPA.