肝癌患者肝组织中螺杆菌感染研究

Status of H. pylori infection in the hepatic tissue of patients with hepatocarcinoma

目的研究肝癌患者肝组织中螺杆菌(Hp)感染状况,进一步揭示Hp感染与肝癌的关系。方法收集正常人、慢性肝炎、肝癌患者肝组织各30例,均经病理证实。(1)在微需氧环境37℃下应用哥伦比亚培养基分离培养肝脏组织中Hp。(2)系列生化反应和速尿素酶检测。(3)免疫组化。(4)应用PCR扩增螺杆菌属16S rRNA基因及序列测定。结果 (1)所有的肝组织培养均未见细菌生长;(2)正常人、慢性肝炎患者肝组织快速尿素酶检测阳性率为0,肝癌为3%;1例肝癌组织硝酸盐试验和过氧化氢酶检测阳性。(3)免疫组化示9例肝癌组织发现Hp,检出率为30.0%。正常肝组织、慢性肝炎患者肝组织中未观察到Hp,与癌组织差异有统计学意义(P<0.05)。(4)PCR扩增产物经1%琼脂糖凝胶电泳,于紫外检测仪下均可见16SrRNA基因阳性扩增带为109bp;30例原发性肝癌标本中有18例Hp16S rRNA基因,正常人、慢性肝炎组均无检出Hp16SrRNA基因,与癌组织差异有统计学意义(P<0.05)。肝组织16S rRNA PCR扩增产物测序对比分析,与Hp16SrRNA片段的基因有98.8%同源性。结论肝癌患者肝组织中存在Hp感染,可能与肝细胞癌存在相关性。

Objective To survey the status of Helicobacter pylori （Hp） infection in the liver of patients with liver cancer and explore the correlation between Hp infection and liver cancer. Methods Liver specimens were obtained 30 patients with chronic liver disease （CLD）, 30 with liver cancer, and 30 healthy control subjects. The specimens were cultured in Colombian agar medium in microaerophilic condition for isolation of Hp. The presence of Hp was also identified by biochemical test, urease test, immunohistochemistry, and PCR and sequence analysis. Results Hp was found in none of the liver specimens by culture tests. One out of the 30 patients with liver cancer （3%） was positive for Hp by urease tests. Immunohistochemical staining identified Hp positivity in 9 liver cancer tissue （30.0%）, but in none of control or CLD cases （P0.05）. PCR detected Hp 16S rRNA in 18 out of the 30 patients with liver cirrhosis （60.0%） and in none of the control or chronic hepatitis cases. Sequencing analysis of Hp 16S rRNA in liver tissue showed a 98.8% homology with the Hp16S rRNA gene. Conclusion A greater incidence of Hp infection in the liver is identified in patients with liver cancer than in healthy subjects and CLD patients, suggesting a potential correlation between Hp infection to liver cancer.