原发性肝癌组织中螺杆菌相关基因的检测

DETECTION OF HELICOBACTER SPECIES RELATED GENES IN LIVER TISSUE OF PRIMARY LIVER CANCER

目的探讨原发性肝细胞癌(HCC)与螺杆菌感染关系。方法选取经病理诊断的38例HCC病人的癌组织作为实验组,28例非肝癌肝病组织、29例胃癌组织、18例结肠癌组织和18例子宫肌瘤组织作对照组。采用聚合酶链反应(PCR)扩增螺杆菌属16SrRNA基因,扩增产物经Southern杂交确认,并进行测序及同源性比较。阳性者再扩增幽门螺杆菌的特异基因26-kDa及相关基因cagA、vacA、rps4、glmM以进一步验证是否为幽门螺杆菌感染。结果38例HCC标本中有27例检出螺杆菌属16SrRNA基因,阳性率为71.1%;29例胃癌标本有23例检出螺杆菌属16S rRNA,阳性率为79.3%,两组比较差异无显著性(P>0.05);与其他对照组(均未扩增出16SrRNA基因)比较差异有显著性(χ2=24.70～33.67,P<0.01)。16S rRNA PCR产物经Southern杂交证实为幽门螺杆菌。序列测定表明,HCC和胃癌组织中的螺杆菌16SrRNA序列与幽门螺杆菌序列有97.8%的同源性。Hp特异相关基因检测结果,26-kDa基因扩增阳性率82.0%;cagA基因扩增阳性率18.3%;glmM基因扩增阳性率15.0%;均未扩增出vacA和rps4基因。结论HCC病人肝组织中存在螺杆菌感染且感染率较高,螺杆菌感染与HCC可能存在相关性。

Objective To investigate the relationship between helicobacter infection and hepato cellular carcinoma （HCC）. Methods This study consisted of 33 HCC patients confirmed pathologically, 28 samples from patients with non-HCO liver-disease tissue, 29 cases with gastric cancer, and 18 with colon cancer. Eighteen cases with uterus myoma served as controls. Helicobaeter species in liver specimens from the studied subjects were examined by polymerase chain reaction （PCR） with Helico hacter specific 26S rRNA primers. Besides, amplified products were identified by Southern hybridization arid sequenced further. The idio-gene 26-kDa and related genes cagA, vacA, rps4 and glmM of positive findings were also detected by PCR to authenticate whether it was HP. Results Twenty-seven of the 38 samples of HCC were positive for Helicobacter 16S rRNA gene, while in 29 gastric cancer samples, 23 were positive. The difference between the two groups was not significant （P）〉0.05）, compared with the other control groups, the difference was significant （x^2=24. 70-33.67,P〈0.01）. The PCR amplified products were identifiled by Southern hybridization. The sequencing showed 97.8% homology of Helicobacter 16S rRNA in tissue of liver and gastric can cer. Conclusion A higher Helicobacter infection rate exists in HCC tissue, which likely correlates with HCC. The liver tissues of HCC patiens with a high infection rate, suggesting an association between Helicobacter infection and HCC.