利用激光捕获显微分离技术和突变分析法检测肺癌病人痰细胞的K-ras和p53基因突变

Analysis of K-ras and p53 Gene Mutations in Sputum Cells of Lung Cancer Patients with Laser Capture Microdissection and Mutation Detection

[目的]探索激光捕获显微分离技术在检测肺癌病人K-ras和p53基因突变的应用。[方法]以3种不同的基因突变分析法对20例患者的痰细胞进行基因突变分析:①以PCR+DGGE方法分析K-ras基因突变,以PCR+SSCP方法分析p53基因突变;②以PCR+MAE+DGGE方法分析K-ras基因突变;以PCR+MAE+SSCP方法分析p53基因突变;③以激光捕获显微分离技术和突变分析法检测肺癌病人痰细胞的K-ras和p53基因突变。[结果]在20例患者中,以现有的方法仅发现有15%(3/20)的基因突变,而激光捕获显微分离技术则发现有60%(12/20)的基因突变。[结论]激光捕获显微分离技术和突变分析法能高效灵敏检测肺癌病人痰细胞的K-ras和p53基因突变。

[Objective] To explore the possibility of application of laser capture microdissection to analyze K-ras and p53 gene mutations in lung cancer patients. [Methods] K-ras and p53 gene mutations in sputum cells of 20 cases were analyzed using 3 different methods respectively：（1） K-ras mutation was analyzed by PCR＋DGGE and p53 mutation by PCR ＋ SSCP. （2） K-ras mutation was analyzed by PCR＋ MAE＋ DGGE, and p53 mutation by PCR＋ MAE＋ SSCP. （3） K-ras and p53 mutations were analyzed by laser capture microdissection and mutation detection. [Results] 15% （3/20） Gene mutations were found using the exist methods. However, 60%（12/20） gene mutations were found using laser capture microdissection and mutation detection. [Conclusion] Comparing to the exist methods, the laser capture microdissection and mutation detection are more sensitive to detect K-ras and p53 gene mutations in sputum cells of lung cancer patients.