cagA基因阳性幽门螺杆菌培养滤液对人胃粘膜上皮细胞系GES-1生长的影响

The effects of cagA positive culture filtrates of Helicobacter pylori to the growth of human gastric epithelial cells

目的 研究cagA+ 幽门螺杆菌 (Hp)培养滤液对人胃粘膜上皮细胞 (GES 1)的作用及机制。方法 制备Hp培养滤液 ,PCR鉴定cagA基因。采用倒置显微镜、电镜、细胞生长曲线、克隆形成实验、单细胞微凝胶电泳及流式细胞仪等 ,观察Hp(cagA+ )培养滤液对GES 1细胞的作用。结果 经Hp(cagA+ )培养滤液处理的GES 1细胞 ,细胞核增大、畸形、核染色质变粗、核仁肥大、核分裂。生长曲线可见细胞增生活跃 ,增殖率 195 %。克隆形成试验显示细胞克隆形成能力增强 ,增殖率达到 337.5 %。流式细胞仪S期细胞比率显著高于对照组。Hp(cagA+ )培养滤液可使GES 1细胞形成彗星现象。结论 Hp(cagA+ )培养滤液可以导致GES 1细胞的生长特性改变 ,呈现肿瘤细胞的形态学及生长特征。DNA损伤可能是cagA诱导GES 1细胞生长特性改变的机制之一。

Objective To study the effects and mechanism of malignant transformation induced by cagA gene positive broth culture filtrates of Helicobacter pylori in human gastric epithelial cells. Methods Broth culture filtrates of Helicobacter pylori were prepared. These filtrates were identified by PCR. GES 1 cells were treated with the filtrates and then examined by microscopy, growth curve, colony forming, single cell microgel electrophoresis and FCM were used. Results GES 1 cells, treated with cagA gene positive broth culture filtrates of Helicobacter pylori showed dyskaryosis, hypertrophy of nucleolus and karyokinesis. The survival rate was 195% by growth curve and 337.5% by colony formation. The cell of S stage cycle was increased compared to control. comet assay showed DNA damage in GES 1 cells by cagA positive filtrates. Conclusion The changes of the morphology and growth character of tumor cells were induced by cagA gene positive broth culture filtrates of Helicobacter pylori in human gastric epithelial cells. DNA damage maybe be one of the mechanisms of the changes in growth character induced by cagA gene positive broth culture filtrates. [