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目的:研究口腔黏膜异常增生不同阶段上皮细胞E-cadherin及Twist1启动子甲基化,探讨其意义。方法:使用量子点免疫荧光技术检测口腔黏膜异常增生中Twist1的表达。随机选取口腔黏膜上皮异常增生组织,使用甲基化特异性PCR检测其甲基化状态,分别以口腔黏膜单纯过度角化,口腔鳞癌细胞系及口腔鳞癌组织作为阴性或阳性对照。结果:Twist1在口腔黏膜单纯过度角化中几乎不表达,在口腔异常增生黏膜中随上皮异常增生程度加重而升高(P<0.05)。在轻、中、重度的口腔黏膜异常增生的样本中,Twist1甲基化率随上皮异常增生程度加重而升高,分别为15.8%,29.4%,50%,差异有显著性(P<0.05)。口腔癌组织Twist1甲基化率为62.5%,口腔癌细胞系为60%,而单纯过度角化中Twist1未出现甲基化;与异常增生相比具有统计学意义(P<0.05)。在不同程度的异常增生上皮中,E-cadherin甲基化率分别为33.3%,50%,62.5%,未显示统计学差异。Twist1和Ecadherin的甲基化率无明显相关性(P>0.05)。结论:Twist1启动子甲基化可能发生在黏膜异常增生早期阶段,且与口腔黏膜上皮癌变过程关系密切。
Objective: To investigate the methylation of E-cadherin and Twist1 promoter in epithelial cells in different stages of oral mucosal dysplasia and to explore its significance. Methods: Quantitative dot immunofluorescence was used to detect the expression of Twist1 in oral mucosal dysplasia. Randomly selected oral mucosa epithelial dysplasia tissues, methylation specific PCR was used to detect the methylation status, oral hyperplasia of oral mucosa, oral squamous cell carcinoma and oral squamous cell carcinoma tissues as negative or positive control. Results: Twist1 was almost not expressed in oral hyperkeratosis and increased in epithelial dysplasia with dysplasia of oral mucosa (P <0.05). In mild, moderate and severe oral mucosal dysplasia samples, the methylation rate of Twist1 increased with the severity of epithelial dysplasia (15.8%, 29.4% and 50%, respectively) (P <0.05) . The methylation rate of Twist1 was 62.5% in oral cancer tissues and 60% in oral cancer cell lines. However, there was no methylation of Twist1 in simple hyperkeratosis, which was statistically significant compared with dysplasia (P <0.05). The methylation rates of E-cadherin were 33.3%, 50% and 62.5% respectively in different degrees of dysplastic epithelium, showing no statistical difference. There was no significant correlation between methylation rates of Twist1 and Ecadherin (P> 0.05). Conclusion: Twist1 promoter methylation may occur in the early stage of mucosal dysplasia, which is closely related to carcinogenesis of oral mucosa.