摘要
目的:研究幽门螺杆菌L型(helicobacter pylori L-form,Hp-L型)感染对胃癌BGC-823细胞增殖的影响,并探讨其作用机制。方法:将胃癌BGC-823细胞与Hp-L型以不同比例(1:20、1:100、1:500)共培养,以不加Hp-L型为对照组,在不同时段进行以下实验:倒置显微镜观察细胞形态学变化;四甲基噻唑氮蓝(MTT)法检测细胞生长增殖率;流式细胞仪(FCM)检测细胞凋亡率;免疫组化(SP法)检测癌基因(skp2)、抑癌基因(p53)和核增殖指数(Ki-67)的表达情况。结果:Hp-L型作用BGC-823细胞后,倒置显微镜观察到细胞分裂增多,增殖旺盛,瘤巨细胞增多,出现明显的生长加速现象;MTT法测定细胞生长曲线显示,Hp-L型对胃癌BGC-823细胞增殖有促进作用;FCM检测可见,Hp-L型影响胃癌BGC-823细胞周期的分布,使G0/G1期比例降低,S期比例升高;免疫组化可见细胞中Skp2、p53和Ki-67蛋白表达阳性率逐渐增加;以上作用均呈细菌浓度和作用时间依赖性。结论:Hp-L型可促进胃癌BGC-823细胞增殖,其机制与上调Skp2、p53和Ki-67蛋白的表达有关。
Objective：To investigate the effect of Helicobacter pylori L - form （ Hp - L） on proliferation of gastric cancer cells BGC - 823 and the probable mechanism involved. Methods： BGC - 823 cells were co - incubated with different proportions of Hp - L （ 1 ： 20,1 ： 100,1 ： 500 ） and the experiments were carried out at different period of time. Morphology changes of BGC -823 ceils were observed by inversion microscope; The proliferous rate of cell growth was assessed by MTT spectrophotometric analysis;The cell cycle was measured by flow cytometry （FCM） ;Immunohistochemical staining was used to detect the expressions of Skp2, p53 and Ki - 67 in BGC - 823 cells. Results： By inversion microscope, BGC -823 cells were identified by their typical cobblestone configuration which adhibited the bottle wall with irregular morphology in normal situation, but when it were infected by Hp - L, division ceils increased and the cells grew bloom, many tumor giant cells could be seen, which indicated the growth acceleration of the cells;The growth curve of cell growth rate determined by MTT method showed that Hp- L could promote the BGC -823 cells proliferation; The FCM showed that Hp -L changed the cell cycle distribution notably and caused a S phase advance , cells at G0/G1 phase were diminished ; Expression of Skp2, p53 and Ki - 67 show that the positive ceils number gradually increased, all the results became more obviously with the increase of concentration of Hp - L and the extention of time in a manner of concentration - and - time - dependent. Concision： Hp - L may promote the proliferation of gastric cancer ceils BGC - 823, its mechanism may be related to up regulation of Skp2, p53 and Ki- 67 expression.
出处
《现代肿瘤医学》
CAS
2011年第2期229-234,共6页
Journal of Modern Oncology
基金
安徽省教育厅高校自然科学研究重点项目(编号:KJ2007A098)
蚌埠市科技局2007年第一批科技项目(编号:11)
