Helicobacter infection in hepatocellular carcinoma tissue

AIM： To investigate whether Helicobacter species （Helicobacter spp.） could be detected in hepatocellular carcinoma （HCC） tissue.METHODS： Liver samples from 28 patients with hepatocellular carcinoma （HCC） diagnosed by histopathology were studied. Twenty-two patients with other liver diseases （5 with liver trauma, 7 with cavernous liver hemangioma, 6 with liver cyst and 4 with hepatolithiasis）, 25 patients with gastric cancer, 15 with colonic cancer and 15 with myoma of uterus served as controls. Two piceces of biopsy were obtained from each patient. One was cultured for Helicobacter spp. and extraction of DNA, the other was prepared for scanning electron microscopy （SEM） and in situ hybridization. The samples were cultured on Columbia agar plates with microaerobic techniques. Helicobacter spp. in biopsy from the studied subjects was detected by polymerase chain reaction （PCR） with Helicobacter spp. 16S rRNA primers. Amplified products were identified by Southern hybridization and sequenced further. Besides, other genes （vacA, cagA） specific for Helicobacter pylori （H pylorO were also detected by PCR. Helicobacter spp. in biopsies was observed by SEM. Transmission electron microscopy （TEM） was performed to identify the cultured positive Helicobacter spp. The presence of Helicobacter spp. was detected by in situ hybridization to confirm the type of Helicobacter. RESULTS： The positive rate of He/icobacter cultured in HCC and gastric cancer tissue was 10.7% （3/28） and 24%（6/25）, respectively. Helicobacter microorganisms were identified further by typical appearance on Gram staining, positive urease test and characteristic colony morphology on TEM. The bacterium was observed in adjacent hepatocytes of the two HCC samples by SEM.The number of cocci was greater than that of bacilli. The bacterium was also found in four gastric cancer samples. PCR showed that the positive rate of HCC and gastric cancer samples was 60.7% and 72% respectively, while the controls were negative （P〈 0.01）. The PCR-amplified products were identified by Southern hybridization and sequenced. The homology to 16S rRNA of H pylon was 97.80%. The samples were verified by in situ hybridization for Helicobacter spp. 16S rRNA-mRNA and proved to be Hpylori positive. There was no statistical significance between HCC and gastric cancer （P〉 0.05）, but the positive rate of HCC and controls had statistical significance （P〈0.01）. Only 3 HCC samples and 2 gastric cancer samples of the cagA genes were detected. None of the samples reacted with primers for vacA in the two groups. As for the genotype of H pylori, type II had preference over type I. CONCLUSION： Helicobacter infection exists in liver tissues of HCC patients. Helicobacter spp. infection is related with HCC, which needs further research.

瞄准：调查 Helicobacter 种类(Helicobacter spp ) 是否能在肝细胞被检测癌(HCC ) 织物。方法：从有组织病理学说诊断的肝细胞癌(HCC ) 的 28 个病人的肝样品被学习。有另外的肝疾病的 22 个病人(5 与肝损伤， 7 与多孔的肝缝血管瘤， 6 与肝包囊并且 4 与肝石病) ，有胃的癌症的 25 个病人， 15 与结肠的癌症并且 15 与子宫的肌瘤用作控制。二活体检视从每个病人被获得。一个人为 Helicobacter spp 是有教养的。并且 DNA 的抽取，其它为扫描电子显微镜学(SEM ) 和原位杂交作好准备。样品在有微氧气的技术的哥伦比亚琼脂平皿上是有教养的。Helicobacter spp。被聚合酶链反应(PCR ) 与 Helicobacter spp 在从学习题目的活体检视检测。16S rRNA 教材。放大产品被南部的杂交识别并且进一步定序。而且，另外的基因(休假 A， cagA ) 为 Helicobacter pylori (H pylori ) 的 specific 被 PCR 也检测。Helicobacter spp。被 SEM 在活体检视观察。传播电子显微镜学(TEM ) 被执行识别有教养的积极 Helicobacter spp。Helicobacter spp 的存在。被原位杂交检测证实 Helicobacter 的类型。结果：在 HCC 和胃的癌症织物有教养的 Helicobacter 的积极的率是 10.7%(3/28 ) 并且 24%(6/25 ) 分别地。Helicobacter 微生物被典型外观在克在 TEM 上染色，积极 urease 和典型殖民地形态学上进一步识别。细菌被 SEM 在二件 HCC 样品的邻近的 hepatocytes 观察。球菌的数字比杆菌的大。细菌也在四件胃的癌症样品被发现。PCR 证明 HCC 和胃的癌症样品的积极的率分别地是 60.7% 和 72% ，当控制是 negative (P<0.01 ) 时。放大 PCR 的产品被南部的杂交识别并且定序。到 H pylori 的 16S rRNA 的相同是 97.80% 。样品被原位杂交为 Helicobacter spp 验证。16S rRNA-mRNA 并且证明是 H pylori 积极。在 HCC 和胃的癌症(P>0.05 ) 之间没有统计意义，但是 HCC 和控制的积极的率有统计意义(P<0.01 ) 。3 件仅仅 HCC 样品和 cagA 基因的 2 件胃的癌症样品被检测。任何一个都没在二个组为休假 A 与教材样品反应。至于 H pylori 的遗传型，类型 II 在类型上有偏爱我。结论：Helicobacter 感染在 HCC 病人的肝纸巾存在。Helicobacter spp。感染与 HCC 被联系，它需要进一步的研究。