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目的:通过对食管癌组织及同一患者对应的术前外周血中RASSF1A基因甲基化的检测,加深食管癌变分子机制的了解并为食管癌早期诊断和高危人群预警提供候选指标。方法:采用甲基化特异性PCR方法,分别检测来自食管癌高发区的30例食管癌患者血浆、肿瘤组织及癌旁正常组织中RASSF1A基因启动子甲基化状况。结果:食管癌组织RASSF1A甲基化阳性率为40%(12/30),而这12例癌组织甲基化阳性的患者其外周血甲基化阳性共7例,癌组织和外周血RASSF1A甲基化阳性一致率为58%(7/12),18例癌组织甲基化阴性的患者其外周血也均为阴性,阴性一致率为100%(18/18)。癌旁正常食管组织甲基化率为13%(4/30),明显低于癌组织(40%,12/30),P<0.05。7例外周血甲基化阳性的患者中,淋巴结转移阳性5例(55%,5/9),阴性2例(10%,2/21),两者差异有统计学意义,P<0.05。低分化鳞癌的RASSF1A甲基化阳性率(83%,5/6)明显高于中分化鳞癌(24%,5/21),P<0.05。结论:外周血RASSF1A甲基化可以反映同一个体食管鳞癌组织中RASSF1A基因甲基化的状态,可能是高危人群筛查重要候选分子标志之一。
OBJECTIVE: To investigate the methylation of RASSF1A gene in esophageal cancer tissue and the corresponding preoperative peripheral blood to further understand the molecular mechanism of esophageal cancer and provide a candidate index for early diagnosis of esophageal cancer and early warning of high-risk population. Methods: Methylation-specific PCR was used to detect the methylation status of RASSF1A promoter in plasma, tumor tissues and adjacent normal tissues from 30 esophageal cancer patients with high esophageal cancer incidence. Results: The positive rate of methylation of RASSF1A was 40% (12/30) in esophageal cancer tissues, while in the 12 cases with methylation-positive methylation, 7 cases were positive methylation of RASSF1A in cancer tissues and RASSF1A in cancer tissues and peripheral blood The positive rate of biochemistry was 58% (7/12). The methylation-negative patients in 18 cases were also negative in peripheral blood. The negative consistent rate was 100% (18/18). The methylation rate of adjacent normal esophageal tissues was 13% (4/30), which was significantly lower than that of the cancerous tissues (40%, 12/30), P <0.05. Of the 7 patients with methylation of peripheral blood, lymph node metastasis There were 5 positive cases (55%, 5/9) and 2 negative cases (10%, 2/21). There was a significant difference between the two groups (P <0.05). The positive rate of RASSF1A methylation in poorly differentiated squamous cell carcinoma (83%, 5/6) was significantly higher than that in moderately differentiated squamous cell carcinoma (24%, 5/21) (P <0.05). Conclusion: Methylation of RASSF1A in peripheral blood can reflect the methylation status of RASSF1A gene in the same esophageal squamous cell carcinoma, which may be one of the important candidate molecular markers for screening at high risk population.