胰腺癌组织K-ras基因第12密码子突变的定量检测

Quantitative detection of mutation of K-ras gene at codon 12

目的检测胰腺癌及相应癌旁组织K—ras基因第12密码子的突变量，分析其与胰腺癌临床病理参数的关系。方法应用肽核酸（peptide nucleic acid，PNA）钳制双荧光标记探针的实时定量PCR法检测93例胰腺癌组织及其癌旁组织中K—ras基因第12密码子的突变拷贝数，以突变百分率表示突变量。K-ras基因突变百分率=K—ras突变型拷贝数／（K—ras野生型拷贝数＋K-ras突变型拷贝数）×100％。结果胰腺癌及其癌旁组织的K-ras基因第12密码子突变率分别为83．9％和65．6％，相差显著（P〈0．05）；它们的突变量分别为（13．385±1．745）％和（2．246±0．728）％，相差亦显著（P〈0．05）。K—ras基因第12密码子突变量与临床病理参数无关。结论胰腺癌及相应癌旁组织不仅存在K-ras基因突变率的差异，亦存在K—ras基因突变量的差异。

Objective To quantitatively analyze the K-ras gene mutation at codon 12 in pancreatic cancer tissues and the relationship between K-ras gene mutation and clinicopathologieal parameters, Methods Quantitative detection of K-ras gene at codon 12 in 93 pairs of pancreatic cancer and adjacent tissues were performed by using PNA-mediated PCR clamping with two different fluorescence labeled probes. The quantity of mutation was expressed by percentage of mutation. The percentage of K-ras gene mutation = the copy of K-ras mutation/（ copy of wild type K-ras ＋ copy of K-ras mutation） × 100%. Results The percentage of mutation of K-ras gene at codon 12 in pancreatic cancer and adjacent tissues were 83.9% and 65.6% , and the difference was statistically significant （P 〈 0.05 ） ; and the quantity of mutation were （ 13. 385 ± 1. 745 ） % and （2.246 ± 0.728 ）%, and the difference was also statistically significant（ P 〈 0.05 ）. The quantity of mutation of K-ras gene at codon 12 was not associated with clinicopathological parameters. Conclusions The percentage of K-ras gene mutation, as well as the quantity of K-ras gene mutation was different in pancreatic carcinoma and adjacent tissues.