原发性肝细胞癌组织中kruppel样因子6基因mRNA突变的测定  

A candidate tumor suppressor gene mutated in primary hepatocellular carcinoma:kruppel-like factor 6

作　　者：王少平[1] 陈孝平[1] 裘法祖[1] 

机构地区：[1]华中科技大学同济医学院附属同济医院肝脏外科中心,武汉430030

出　　处：《中华外科杂志》2004年第20期1258-1261,共4页Chinese Journal of Surgery

基　　金：卫生部临床学科重点资助项目 [2 0 0 1( 3 2 1) ]

摘　　要：目的　探讨kruppel样因子 (KLF) 6基因与原发性肝细胞肝癌发生、发展的关系。方法分别用逆转录 聚合酶链反应 (RT PCR)技术和聚合酶链单链构象多态分析 (PCR SSCP)技术、DNA序列测定方法 ,检测 2 7例肝细胞癌患者的癌组织与癌旁组织、5例肝转移癌患者的正常肝组织中KLF6基因mRNA的表达及突变情况。结果　正常肝组织及癌旁组织中 ,只有 1例KLF6mRNA无表达 ,阳性率 97% (31/ 32 ) ;肝细胞癌组织中 ,有 4例无表达 ,阳性率 85 % (2 3/ 2 7) ,两者表达率间差异无显著性(χ2 =2 5 8,P >0 0 5 )。 2 7例肝细胞癌组织中 ,6例出现DNA片段异常泳动带 ,突变率为 2 2 % (6 / 2 7) ;14例出现信息个体 ,其中 5例 (5 / 14 ,36 % )为杂合子缺失 ,这 5例中有 3例出现DNA片段异常泳动带 ,突变比例为 3/ 5。在 2 7例癌旁组织中 1例检测出突变 ,但在检测出突变的肝癌组织的配对癌旁组织中无一例检测出突变。结论　肝细胞肿瘤存在KLF6基因突变 。Objective To explore the relationship between kruppel-like factor (KLF)6 gene and the development or progression of hepatocellular carcinoma(HCC). Methods Reverse-transcription polymerase chain reaction (RT-PCR) was used to examine the expression of KLF6 mRNA in normal liver tissue and primary hepatocellular carcinoma, and single strand conformation polymorphism (SSCP), DNA sequencing were used to detect the point mutation of KLF6 in primary hepatocellular carcinoma. Results An amplified fragment of 427 bp DNA was detected in 31 (97 % ) of 32 adjacent noncancerous tissue and nomal liver tissue, and in 23 (85%) of 27 HCCs. There was no significant difference in the levels of KLF6 mRNA between normal liver and liver tumors (χ~2=2.58,P>0.05). For the 27 HCCs, six SSCP-positive bands (22 %) were detected. Among them, three of 5 (3/5) tumor samples showing loss of heterozygosity(LOH) of KLF6 had mutations in the retained KLF6 allele. Conclusion We showed that LOH was detected in 5 (36 %) HCCs obtained from 14 informative cases, and three of 5 tumor samples showing LOH of KLF6 had mutations in the retained KLF6 allele. Two inactivating events had occurred; thus, as defined by Knudson′s ”two-hit model”, 16 KLF6 appears to be a tumor suppressor gene.

关 键 词：突变 癌旁组织 基因 RNA 表达 癌组织中 肝细胞癌组织 DNA片段 DNA序列 体细胞 

分 类 号：R735.7[医药卫生—肿瘤] R737[医药卫生—临床医学]