不同幽门螺杆菌临床菌株对人胃黏膜细胞系GES-1增殖和凋亡的影响及其致癌性的研究

摘要

Objective To explore the effect of different Helicobacter pylori (H.Pylori) clinical strains on the proliferation and apoptosis of gastric epithelial cells, and to observe the effect of H. Pylori on gastric mucosa by Mongolian gerbil model infected H. Pylori. Methods H. Pylori isolates harvested from pathologically documented gastric carcinoma (GC, n - 10) or chronic gastritis specimens (CG,n= 10) were co-cultured with GES-1 cells individually. MTT assay and flow cy-tometry were used to determine the proliferation and apoptosis of GES-1 cells induced by H. Pylori I-solates.Mongolian gerbils were infected by the most (A strain) and the least (B strain) significant-ly proliferated H. Pylori strains. Results When co-cultured with the cell/bacteria concentration ratio 1:1 and 1: 50 for 12 h and the cell/bacteria concentration ratio 1: 50 for 24 h, H. Pylori clinical strains isolated from patients with gastric cancer promoted the proliferation of GES-1 cells, and there was significant difference in the absorbance compared with the group of gastritis strains (P <0.05). The apoptosis rate of the GC and CG groups increased significantly ( P < 0. 05 ) compared with the control group when co-cultured with the cell/bacteria concentration ratio 1: 50 and 1: 200, and there was no significant difference between the GC group and the CG group (P>0. 05). The incidences of intestinal metaplasia and dysplasia in the A strain group were significantly higher than those in the B strain group (P < 0.05). Conclusion H. Pylori strains from different disease sources have different effects on the proliferation of GES-1 cells. H. Pylori isolated from gastric cancer can promote the proliferation of cells to different degrees and directly induce gastric precancerosis and gastric cancer.%目的:比较不同幽门螺杆菌(H.pylori)临床菌株对人胃黏膜细胞系GES-1细胞增殖、凋亡的影响,了解不同毒力菌株对胃黏膜的影响及致癌性.方法:选用临床分离自胃癌、胃炎患者的菌株各10例分别与GES-1细胞以不同浓度比例共培养,在不同时间点进行MTT和流式细胞仪检测,筛选出对GES-1细胞的增殖影响最大(A菌株)和最小(B菌株)的两株菌株,利用蒙古沙土鼠感染模型了解其导致胃黏膜病变情况及致癌性.结果:当细胞/细菌浓度比为1∶1,1∶50共培养12 h和细胞/细菌浓度比为1∶50共培养24h时,分离自胃癌患者的H.pylori临床菌株可以不同程度地促进GES-1细胞增殖,与胃炎菌株组的吸光度值比较差异有统计学意义(P＜0.05).当细胞/细菌浓度比为1∶50,1∶200共培养时,胃炎菌株组和胃癌菌株组细胞凋亡率较对照组明显增加(P＜0.05),胃炎菌株组与胃癌菌株组间差异无统计学意义(P＞0.05).筛选出的2株菌株感染蒙古沙土鼠模型后28周,A菌株引起蒙古沙土鼠胃黏膜肠化及癌变,与B菌株组比较差异有统计学意义(P＜0.05).结论:不同来源的H.pylori临床菌株对GES-1细胞的增殖影响具有差异性,分离自胃癌黏膜的菌株可明显促细胞增殖,并可以引起蒙古沙土鼠严重的胃黏膜癌前病变及胃癌发生.