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目的 探讨在食管拉网细胞中进行食管癌基因检测的方法及可行性 ,了解食管癌前期病变细胞中p5 3基因的突变与癌变的关系。方法 对四川省盐亭县 1982年进行食管癌普查的食管拉网脱落细胞涂片标本 ,应用聚合酶链反应 -单链构象多态性分析 (PCR SSCP)方法 ,检测其p5 3基因第 5外显子及第 7外显子的突变情况。结果 全组 4 8例标本中 ,食管正常上皮和重度不典型增生上皮各 2 4例 ,p5 3基因检测均获成功。食管重度不典型增生上皮细胞中有 5例检测到突变 ,均为p5 3基因第 7外显子突变 ,而第 5外显子未检测到突变 ;正常食管鳞状上皮拉网脱落细胞中未发现突变。检测到突变的 5例食管上皮重度不典型增生者 ,有 3例分别在 10年、12年和 14年后转变为食管癌。结论 p5 3基因突变的食管上皮不典型增生细胞具有明显的癌变趋势 ;对食管拉网脱落细胞涂片标本进行微量DNA的提取、扩增和基因检测 ,可作为研究食管癌前期病变的一种新方法。
Objective To investigate the method and feasibility of esophageal cancer gene detection in esophageal net cells and to understand the relationship between the mutation of p5 3 gene and canceration in esophageal precancerous lesion cells. Methods A smear of esophageal exfoliated cells in esophageal cancer screening in 1982 in Yanting County, Sichuan Province was detected by polymerase chain reaction-single strand conformation polymorphism (PCR SSCP) Exon and exon 7 mutations. Results Forty-four specimens from all the patients in our study showed 24 cases of esophageal normal epithelium and severe atypical hyperplasia epithelium respectively. The detection of p5 3 gene was successful. Esophageal atypical hyperplasia epithelial cells in 5 cases were detected mutations were p5 3 gene exon 7 mutations, and exon 5 mutations were not detected; normal esophageal squamous cell detachment cells found no mutation. Three of the five patients with esophageal dysplasia who detected a mutation had esophageal cancer at 10, 12, and 14 years, respectively. Conclusion The esophageal epithelial atypical hyperplasia cells with p5 3 mutation have obvious tendency of carcinogenesis. DNA extraction, amplification and gene detection of esophageal exfoliated cells can be used as a new method to study the precancerous lesions of esophageal carcinoma method.