螺杆菌感染与NF—κB p65蛋白表达相关性在人食管癌发生中的意义

The relationship of NF-κB p65 protein expression with Helicobacter species infection and carcinoma of esophagus in humans

目的研究人食管癌是否存在螺杆菌感染及其与NF-κB p65蛋白的关系，从而探讨在人食管癌发生中的意义。方法随机收集食管癌标本40例和正常食管标本10例，采用聚合酶链反应（PCR）扩增螺杆菌16SrRNA，随机选4例测序及同源比较。阳性者再扩增幽门螺杆菌（H.pylari,Hp）的特异基因相对分子质量（Mr）26×10^3的种特异性抗原和相关功能基因（GagA，VacA）。并采用免疫组化法检测食管癌组织中NF-κB p65蛋白的表达情况。结果32．5％（13／40）的食管癌组织中发现有螺杆菌16SrRNA基因存在，正常人食管组织无一例阳性。4例测序及同源比较，食管癌组织中螺杆菌序列与H．pylori的16SrRNA序列有99％～100％同源性。阳性标本均扩增出脚特异基因M，26×10^3种特异性抗原，但仅2例扩增出G田弘基因，3例扩增出VacA基因。免疫组化法染色显示，16SrRNA阳性的食管癌组织中NF-κB p65蛋白表达率为100％（13／13），而16SrRNA阴性食管癌组织表达率仅占18．5％（5／27），二者差异有统计学意义（P〈0．01）。结论食管癌患者食管组织存在螺杆菌感染，该螺杆菌可能为脚。螺杆菌感染导致食管黏膜NF-κB p65蛋白表达的增加，可能在食管癌生成中作为信号传导机制之一。

Objective To investigate the significance of relationship of NF-κB p65 protein expression with Helicobacter species infection in human esophageal carcinoma. Methods Fourty esophageal carcinoma specimens and 10 normal esophageal specimens from surgical resection were randomly collected and studied. Helicobacter species were examined by polymerase chain reaction （PCR） using Helicobacter-specific 16S rRNA primers. A search for other genes that were specific for Helicobacter pylori（ H. pylori） was carried out by PCR. The immanohistochemistry staining for NF-κB p65 protein was performed. Results Helicobacter species were found in esophageal tissue in 32.5%（13/40） patients with squamous carcinoma in which H. pylori-specific gene 26×10^3 SSA in all the 16S rRNA-positive samples were positively amplified by PCR while no positive result was recorded in control group（ P 〈0.05）. Only 2 samples reached with primers for CagA, and 3 samples reached for VacA. Four of Helicobacter-specific PCR amplicons were sequenced and the homology was 99%-100% with 16S rRNA of H. pylori. NF-κB p65 protein was positive in 100% （13/13） Helicobacter 16S rRNA-positive esophageal squamous carcinoma and in 18.5% （5/27） in Helicobacter 16S rRNA-negative esophageal squamous carcinoma（ P 〈 0.01 ）. Conclusion Helicobacteraceae are identified in human esophageal carcinoma tissues with infection of Helicobacter species likely H. pylori. NF-κB p65 protein expression increases markedly in 16S rRNA-positive carcinoma tissues, which might be one of mechanisms in signal transmission in human esophageal carcinoma generation and development.