Xu Ying, Yan Ying, Hou Ming-Xiao, Liu Yun-En
Radiation oncology Department of General Hospital of Shenyang Military Command, Shenyang, l10016 China.
Emergency Medicine Department of General Hospital of Shenyang Military Command, Laboratory of Rescue Center of Severe Wound and Trauma PLA, 83 Wenhua Road, Shenhe District, Shenyang, l10016 China.
Infect Agent Cancer. 2015 Mar 1;10:8. doi: 10.1186/s13027-015-0003-3. eCollection 2015.
Both H. pylori infection and high salt (NaCl) diet are risks of gastric cancer, however, the interaction pattern of the two is not very clear. Our objective was to investigate the effects of NaCl-pretreated H. pylori on DNA damage and proliferation of gastric epithelial cell (GES-1).
GES-1 cells were co-cultured with H.pylori or NaCl-pretreated H. pylori (with 30% NaCl) for 24 h. The morphological changes of all cells were observed by inverted phase contrast microscopy and transmission electron microscopy. Oxidative DNA damage was examined by immunofluorescence. Alterations in mitochondrial membrane potential and apoptosis rate were detected by flow cytometry and western blot, and expression of Ki-67, PCNA and P21 were evaluated using the immunocytochemical staining.
GES-1 cells co-cultured with NaCl-pretreated H.pylori exhibited morphological changes and oxidative DNA damage. Although no significant disruption of the mitochondrial membrane potential (ΔΨm) and apoptotic rate were observed compared with control groups, there were significant decreased in Bax and Caspase3 proteins and increased in Bcl-2 protein in GES-1 cells infected with H. pylori (30) when compared with GES-1 cells cultured with H. pylori. In addition, we found a proliferative effect on GES-1 cells with an increased expression of Ki-67 and PCNA as well as a decreased p21 expression, through which the cells may acquire the potential for malignant transformation.
NaCl-pretreated H. pylori possessed the ability to cause cell injury and promote proliferation in gastric epithelial cells.
幽门螺杆菌感染和高盐（氯化钠）饮食均为胃癌的风险因素，然而，二者的相互作用模式尚不完全明确。我们的目的是研究经氯化钠预处理的幽门螺杆菌对胃上皮细胞（GES-1）DNA损伤和增殖的影响。
将GES-1细胞与幽门螺杆菌或经30%氯化钠预处理的幽门螺杆菌共培养24小时。通过倒置相差显微镜和透射电子显微镜观察所有细胞的形态变化。通过免疫荧光检测氧化性DNA损伤。采用流式细胞术和蛋白质印迹法检测线粒体膜电位和凋亡率的变化，并使用免疫细胞化学染色评估Ki-67、PCNA和P21的表达。
与经氯化钠预处理的幽门螺杆菌共培养的GES-1细胞呈现出形态变化和氧化性DNA损伤。尽管与对照组相比，未观察到线粒体膜电位（ΔΨm）和凋亡率有明显破坏，但与用幽门螺杆菌培养的GES-1细胞相比，感染幽门螺杆菌（30）的GES-1细胞中Bax和Caspase3蛋白显著减少，Bcl-2蛋白增加。此外，我们发现对GES-1细胞有增殖作用，Ki-67和PCNA表达增加，p21表达降低，通过这种方式细胞可能获得恶性转化的潜能。
经氯化钠预处理的幽门螺杆菌具有导致胃上皮细胞损伤和促进其增殖的能力。
