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目的:研究唾液腺腺样囊性癌(adenoid cystic carcinoma,ACC)中RASSF1A表达及其与启动子区甲基化之间的关系。方法:收集167例原发性唾液腺ACC,亚硫酸盐测序聚合酶链反应(bisulfite sequencing polymerase chain reaction,BSP)和甲基化特异性聚合酶链反应(methylation-specific polymerase chain reaction,MSP)方法检测RASSF1A基因启动子区甲基化状况,免疫组织化学方法检测RASSF1A蛋白表达情况。去甲基化药物decitabine处理ACC细胞系SACC-83后,检测处理前、后RASSF1A基因甲基化及表达情况。应用SPSS18.0软件包对数据进行统计学分析。结果:59/167(35.3%)例病例中检测到RASSF1A基因启动子区甲基化。101/167(60.5%)例病例中RASSF1A蛋白呈低或不表达,66/167(39.5)病例中RASSF1A蛋白呈高表达。存在RASSF1A基因甲基化组,RASSF1A蛋白表达显著低于不存在甲基化组(P=0.012)。去甲基化药物decitabine处理ACC细胞系后,RASSF1A mRNA及蛋白水平表达均升高。结论:唾液腺ACC中,启动子区甲基化是RASSF1A基因失活的主要原因,可作为该肿瘤的潜在治疗靶点。
Objective: To investigate the relationship between RASSF1A expression and promoter methylation in adenoid cystic carcinoma (ACC). Methods: 167 cases of primary salivary gland ACC, bisulfite sequencing polymerase chain reaction (BSP) and methylation-specific polymerase chain reaction (MSP) were detected RASSF1A gene promoter methylation status, immunohistochemical detection of RASSF1A protein expression. Demethylation drug decitabine treatment ACC cell line SACC-83, before and after treatment to detect RASSF1A gene methylation and expression. SPSS18.0 software package for statistical analysis of the data. Results: Methylation of promoter region of RASSF1A gene was detected in 59/167 (35.3%) cases. RASSF1A protein was low or not expressed in 101/167 (60.5%) cases and high expression of RASSF1A protein in 66/67 (39.5) cases. There was RASSF1A gene methylation group, RASSF1A protein expression was significantly lower than the absence of methylation group (P = 0.012). Demethylation drug decitabine treatment of ACC cell lines, RASSF1A mRNA and protein levels were elevated. CONCLUSION: Promoter methylation in ACC of salivary gland is the main reason of inactivation of RASSF1A gene, which may serve as a potential therapeutic target for this tumor.