应用PCR-SSCP方法检测非小细胞肺癌患者血浆p53基因突变  

Inspecting the Rate of P53 Mutation in Plasma of NSCLC by PCR-SSCP

作　　者：于正洪[1] 林勍[1] 林勇[1] 刘畅[1] 

机构地区：[1]南京军区南京总医院肿瘤科,江苏南京210002

出　　处：《现代检验医学杂志》2006年第2期4-7,共4页Journal of Modern Laboratory Medicine

摘　　要：目的探讨应用聚合酶链反应（PCR）-单链构象多态性分析（SSCP）检测非小细胞肺癌（NSCLC）患者血浆循环核酸p53基因突变检测的可行性,研究NSCLC患者血浆p53基因突变对NSCLC发生、发展和预后估计的意义.方法应用PCR扩增p53基因外显子5～8,并用SSCP分析产物突变情况,再与各生物参数进行相关研究.结果 10例健康人血浆循环核酸p53基因突变检测结果均为阴性,60例NSCLC患者血浆中p53基因5～8外显子突变测定的结果显示有28例出现突变,占47%（28/60）.p53基因突变率与术后肿瘤残留、对治疗的反应显著相关（P＜0.05）,与临床分期、年龄、性别、病理分型无关（P＞0.05）.结论血浆循环核酸p53基因突变对NSCLC的早期诊断、病情监测、预后评估都具有指导意义.PCR-SSCP分析简便、可靠,可作为检测大量标本的突变的可行性方法.Objective By exploring the possibility and the rate of P53 mutation in plasma of NSCLC by PCR-SSCP, discuss the association with the clinical pathological character and evalue its role in the ocurrence,progression and prognosis of NSCLC. Methods Extraction of DNA fragments from 60 NSCLC patients plasma and 10 nomal person plasma were amplified by PCR. The mutation rate of exon 5 to 8 of P53 were analyzed by single-stranded conformation polymorphism （SSCP）. The association was discussed with other biological mark after the result was managed by the SPSS10. 10. Results The rate of p53 gene exon 5 to 8 mutation was significantly higher in NSCLC group plasma （46. 7% ,28/60）than those in the control group plasma （0%, 0/10）. There were statistically significant to 28 mutation was significantly higher in NSCLC group plasma （46. 7% ,28/60）than those in the control group plasma （0% ,0/10）. There were statistically significant between p53 gene mutation and clinical stages,metastasis,cure degree （P〈0. 05）,but no significant association was identified with pathological type,age and sex. Conclusion The plasma p53 gene mutations is a free event in NSCLC patients,which is associated with the clinical stage and metastasis of NSCLC. so it would play an important role in diagnosis,progression and prognosis of NSCLC as a new tumor marker which can be dectected easily by PCR-SSCP.

关 键 词：PCR-SSCP 非小细胞肺癌(NSCLC) 循环核酸 P53基因突变 

分 类 号：R730.261[医药卫生—肿瘤] Q754[医药卫生—临床医学]