应用SSCP-PCR技术研究非小细胞肺癌p53基因突变  

Detection of p53 gene mutations by SSCP-PCR in human non-small cell lung carcinomas

作　　者：王晓玫[1] 成志强[1] 陶凤华[1] 苏学劲[1] 王玲[1] 

机构地区：[1]暨南大学医学院附属二院病理科,广东深圳518020

出　　处：《第四军医大学学报》2003年第18期1650-1652,共3页Journal of the Fourth Military Medical University

基　　金：国家人事部科技择优项目 (1 9990 1 2 2 ) ;深圳市卫生科技计划重点攻关项目 (1 9980 50 1 1 )

摘　　要：目的 :探讨非小细胞肺癌 (NSCLC) p5 3基因突变与临床病理及预后的关系 ,评价检测 p5 3基因点突变作为基因标记物在判断患者预后中的意义 .方法 :应用聚合酶链反应 单链构象多态性 (PCR SSCP)分析技术 ,检测 31例NSCLC及5例肺良性病变石蜡包埋标本 p5 3基因的点突变 .结果 :肺癌组织 p5 3基因点突变率为 4 8% (15 / 31) ,肺良性病变未检出该基因突变 ,两组相比有显著性差异 .各组织学类型肺癌p5 3基因 5～ 8外显子点突变检出频率为 :鳞状细胞癌 9例(9/ 16 ) ,腺癌 3例 (3/ 10 ) ,肺泡细胞癌 1例 (1/ 3) ,大细胞癌 2例 (2 / 2 ) .肺腺癌组织 p5 3基因点突变率比鳞状细胞癌低 .Ⅲ Ⅳ期肺癌点突变率明显高于Ⅰ期肺癌 (P <0 .0 5 ) ,p5 3基因点突变与临床分期有关 ,在低分化肺癌其突变频率明显高于分化好的肺癌 (高分化和中分化肺癌 ) ,p5 3基因突变率与年龄、性别、肿瘤大小和组织学类型无相关性 .结论 :应用PCR SSCP技术检测 p5 3基因突变能够反映肺癌的进展情况、临床分期及肿瘤分化程度 ,是判断肺癌预后的重要生物学指标 .AIM: To investigate the relationship between point mutation in p 53 gene and the clinical pathology as well as the prognosis of human non small cell lung carcinoma (NSCLC) and to assess the prognostic value of this genetic marker in prognosis. METHODS: DNA preparations from paraffin embedded tissues of benign pulmonary diseases ( n =5) and of NSCLC tumor tissue specimens ( n =31) were detected for mutations in p53 exons 5 8 by using polymerase chain reaction single strand conformation polymorphism (PCR SSCP) assay. RESULTS: Detected mutation rate of 48% (15/31) in the pulmonary carcinoma group was significantly higher than that of the control group (0/5). In pulmonary carcinoma group, the mutation rates in p 53 exons 5 8 for squamous cell carcinomas, adenocarcinomas, bronchioloalveolar carcinomas and large cell carcinomas were nine cases (9/16), three cases (3/10), one cases (1/3) and two cases (2/2), respectively. The incidence of p 53 point mutations in adenocarcinomas was lower than that in squamous cell carcinomas. The extent of p 53 gene mutations was associated with stages of TNM classification, with higher mutation rate in p 53 gene in cancer tissues at stages Ⅲ Ⅳ than that at stagesⅠ ( P <0.05). In poorly differentiated tumors, the mutation rates were obviously higher than those in well and moderately differentiated cases. However, the statistical analysis showed no significant correlation between the p 53 mutation and other clinical parameter such as age, sex, tumor size and histological subtype. CONCLUSION: Detection of p 53 gene mutations in lung carcinomas by PCR SSCP can be used as a reliable follow up genetic marker for the early detection or prognostic surveillance of human pulmonary carcinomas.

关 键 词：肺肿瘤 预后 多态现胆 单链构象 聚合酶链反应 基因 p53 

分 类 号：R734.2[医药卫生—肿瘤]