肝癌组织中螺杆菌16SrRNA基因的检测  

Detection of Helicobacter species 16Sr RNA gene in paraffin-embedded hepatocellular carcinoma tissues

作　　者：黄燕[1] 范学工[1] 陈永平[2] 李宁[3] 汤立军 

机构地区：[1]中南大学湘雅医院传染科,湖南省长沙市410008 [2]中南大学湘雅医学院病理教研室,湖南省长沙市410078 [3]中南大学湘雅医院中心实验室,湖南省长沙市410008 [4]中南大学湘雅医学院分子生物研究中心,湖南省长沙市410078

出　　处：《世界华人消化杂志》2002年第8期877-882,共6页World Chinese Journal of Digestology

基　　金：国家教育部骨干教师基金(教技司2000-65);湖南省自然科学基金(01jjy2114);湖南省卫生厅科研基金(00022)资助~~

摘　　要：目的:探讨螺杆菌感染是否与人类肝癌的发生相关.方法:选取经病理诊断的肝癌组织38例,肝硬化15例,慢性肝炎12例,肝良性肿瘤15例,采用聚合酶链反应(PCR)扩增螺杆菌16SrRNA检测螺杆菌,扩增产物经Southern杂交证实,并对部分标本的PCR产物进行测序及同源比较.阳性者再扩增幽门螺杆菌(Hpylori)的特异基因(26Ku蛋白)和相关功能基因(cagA,vacA,glmM,rps4)来验证是否为该菌.结果:52.6%(20/38)的肝癌组织中发现螺杆菌存在,而非肝癌组无一例阳性(P<0.01).所有PCR产物用South-ern杂交得到证实.10例测序及同源比较显示,肝癌组织中的螺杆菌16SrRNA序列与Hpylori的16SrRNA序列有99-100%的同源性.阳性标本中26ku基因大部分亦阳性(14/20),但仅3例扩增出cagA基因,4例扩增出glmM基因,一直未扩增出vacA基因和rps4基因.结论:肝癌组织中存在螺杆菌感染,他与肝癌的发生可能存在某种联系.AIM: To investigate the relationship between Helicobacterspecies and primary tissue carcinoma.METHODS:Liver samples from 38 patients with primaryliver carcinoma diagnosed by histopathology and 15 withliver cirrhosis, 12 with chronic hepatitis, 15 with benignneoplasm as controls were studied. Helicobacter species inliver speimens from studied subjects were examined by poly-merase chain reaction (PCR) using Helicobacter-specific 16SrRNA primers. Amplified products were identified by South-ern blot and sequenced. A search for other genes that werespecific for Helicobacter pylori was also carried out by PCR.RESULTS:Helicobacter-specific 16Sr RNA gene was posi-tive in 52.6% (20/38) samples of hepatocellular carcinoma,while none was positive in control groups (P <0.01). Ten ofHelicobacter-specific PCR amplicons were sequenced andhomology similarity was 99-100% with 16Sr RNA of H pylori.Of these 20 positive samples, 14 were positive with the26ku protein primers of H pylori. Only 3 samples reactedwith primers for cagA, and 4 samples reacted for glmM. Noneof the samples reacted with primers for vacA and rps4.CONCLUSION:Helicobacter species can be present in theliver of patients with hepatocelluar carcinoma and it is prob-ably linked to carcinogenesis of liver mialignance.

关 键 词：肝癌 螺杆菌 16S RRNA基因 聚合酶链反应 相关性 

分 类 号：R735.7[医药卫生—肿瘤]