论文部分内容阅读
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 investigate the effect and mechanism of culture filtrate of cagA + Helicobacter pylori (Hp) on human gastric mucosal epithelial cells (GES 1). Methods Hp culture filtrate was prepared and cagA gene was identified by PCR. The effects of Hp (cagA +) culture filtrate on GES-1 cells were observed by inverted microscope, electron microscope, cell growth curve, clonogenic assay, single-cell microgel electrophoresis and flow cytometry. Results The GES 1 cells treated with Hp (cagA +) culture filtrate showed enlarged nucleus, deformity, thickening of chromatin, hypertrophy of nucleolus, and mitosis. Growth curve shows that cell proliferation is active, the proliferation rate of 195%. Clone formation assay showed that the ability of cell clone formation was enhanced and the proliferation rate reached 337.5%. Flow cytometry S-phase cell ratio was significantly higher than the control group. Hp (cagA +) culture filtrate causes GES 1 cells to form a comet phenomenon. Conclusions Hp (cagA +) culture filtrate can change the growth characteristics of GES 1 cells, showing the morphological and growth characteristics of tumor cells. DNA damage may be one of the mechanisms of cagA-induced changes in the growth characteristics of GES 1 cells.