胆管癌基因变异与临床病理学特征的关系

目的：了解肝内胆管癌（ICC）在发生和发展过程中基因变异特点及其与临床病理学特征的关系。方法：采用微量组织切割法,从22份ICC石蜡包埋组织块中提取基因组DNA,经PCR扩增,琼脂糖凝胶电泳分离和DNA直接测序,检测6种肿瘤抑制基因（APC,MCC,DCC,OGG1,p53和RB1）的杂合性缺失和Ki-ras-2癌基因点突变的状况,分析其与临床病理学指标的关系。结果：7 肿瘤基因变民的总检出率为86．4％（19／22）,根据变异基因类型与临床病理学持征的关系。将19例有基因变异的病例分为两组,I型患者9例,47．4％,具有APC,MCC,DCC和Ki-ras-2基因变异,平均年龄57．2岁,II型患者10例,52．6％,具有P53,OGG1和RB1基因变异,平均年龄69．1岁（P＜0．05）,I型患者的术后3年生存率为88．9％（8例）,明显高于II型患者的30．0％（3例,P＜0．05）,结论：ICC的发生和发展与多基因变异的长期蓄积和协同作用密切相关,I型基因变异（APC,MCC,DCC和Ki-ras-2)主要在ICC的启动和早期学进阶段起作用,II型基因变异P53,OGG1和RB1）则在促进ICC的晚期演进过程中发挥作用,对ICC基因变异谱系的检测有助于了解肿瘤演进状态和判断预后。

Relationship between genetic alterations and clinicopathological features in intrahepatic cholangiocarcinoma

OBJECTIVE: To evaluate the pedigree of genetic alterations during the tumorigenesis of intrahepatic cholangiocarcinoma (ICC) and their correlation with clinicopathological features by analysis of loss of heterozygosity (LOH) in 6 tumor suppressor genes (APC, MCC, DCC, OGG1, p53 and RB1) and point mutations in Ki-ras-2 oncogene. METHODS: Genomic DNA was isolated from paraffin-embedded slides of 22 surgically resected ICC cases by microdissection-based PCR amplification and agarose gel electrophoresis. Genetic alterations were analyzed by direct DNA sequencing. RESULTS: The total frequency of alterations in 7 genes studied was 86.4% (19/22). Based on the pattern of altered genes and their correlation with clinicopathological parameters, the genetic alterations were classified into two groups: Group I (9/19, 47.4%): alterations in APC, MCC, DCC and Ki-ras-2,); Group II (10/19, 52.6%): alterations in p53, OGG1 and RB1. The average age of patients in Group I (mean age, 57.2 years) was significantly younger than those in Group II (mean age, 69.1 years) (P < 0.05). CONCLUSIONS: The occurrence and development of ICC was closely related with the accumulation and cooperation of multiple genetic alterations. The genetic alterations of APC, MCC, DCC and Ki-ras-2 may play crucial roles in the early stage of development of ICC, and the genetic alterations of p53, OGG1 and RB1 may play important roles in accelerating advanced progression of ICC. The detection of the pedigree of genetic alterations in ICC may provide useful information for evaluating the state of tumor progression and clinic prognosis.