幽门螺杆菌对肝癌细胞HepG2的体外细胞毒作用  

In Vitro Cytotoxicity of Helicobacter pylori on Hepatocarcinoma HepG2 Cells

作　　者：陈仁[1] 范学工[1] 黄燕[1] 李宁[2] 陈朝晖[2] 

机构地区：[1]中南大学湘雅医院传染科,湖南长沙410008 [2]中南大学湘雅医院医学研究中心,湖南长沙410008

出　　处：《癌症》2004年第1期44-49,共6页Chinese Journal of Cancer

基　　金：国家自然科学基金(30271171);国家教育部优秀骨干教师基金(教技司2000-65);湖南省自然科学基金(01JJY2114);湖南省卫生厅科研基金(00022)~~

摘　　要：背景与目的:幽门螺杆菌(Helicobacterpylori,HP)是慢性胃炎和消化性溃疡的重要病因,人和动物胃肠外疾病亦与HP感染密切相关,本研究探讨HP对肝癌细胞株HepG2的形态和生长的影响。方法:利用体外细胞培养技术,将不同浓度的cagA(+)HP和cagA(-)HP与HepG2共同培养,应用细胞形态学观察、DNA电泳、透射电镜、MTT检测、酶学检测等技术,观察HP菌液对体外培养HepG2细胞的形态、生长速度和酶学改变的影响。结果:(1)随着cagA(+)HP菌液浓度的增大和作用时间的延长,HepG2细胞形态由贴壁的梭形变为圆形,贴壁细胞减少,悬浮细胞增多,并且细胞周围出现碎片;(2)随着cagA(+)HP菌液浓度的增大,可见DNA条带;(3)随着cagA(+)HP菌液浓度的增大和作用时间的延长,HepG2细胞核染色质浓缩,呈新月形聚集于核缘,胞质浓缩,可见空泡;(4)随着cagA(+)HP菌液浓度的增大,对HepG2细胞杀伤率逐渐增强(P<0.01);(5)与cagA(+)HP菌液共同孵育后,HepG2细胞出现ALT、LDH升高(P<0.01);(6)未发现cagA(-)HP对陈仁,等.幽门螺杆菌对肝癌HepG2细胞生长的影响。结论:cagA(+)HP对HepG2细胞具有明显的体外细胞毒作用,其毒性作用与HP菌液的浓度和作用时间呈明显正相关。BACKGROUND &OBJECTIVE: Helicobacter pylori (HP) is the important p athogen of chronic gastritis and peptic ulcer. HP infection is closely associate d with extragastrointestinal disease of human and animals. This study was design ed to investigate the effect of HP on hepatocarcinoma HepG2 cells. METHODS: HepG 2 cells was cocultured with cagA (+) HP and cagA() HP at different concentrat ions. Then the cell morphology, DNA fragments electrophoresis, transmission elec tron microscopy, MTT method, and enzyme assays were used to observe the effect o f HP on the morphology, growth rate, and enzyme change of HepG2 cells. RESULTS: (1) With the increasing concentrations of cagA (+) HP and prolongation of cultu re time, the morphology of HepG2 cells was changed from spindle shape to round; the ability of HepG2 cells adhering to wall were decreased; the ability of HepG2 cells in suspension were increased; and debris emitted around the cells; (2) Wi th the increasing concentrations of cagA (+) HP, DNA ladder occurred; (3) With the increasing concentrations of cagA (+) HP and prolongation of culture time, the nuclei of HepG2 cells showed chromatin pyknosis, and clustered on the inner border of karyotheca, condensed cytoplasm with many vacuoles; (4) The MTT values of HepG2 cells were decreased with the increasing concentration of cagA(+) HP (P< 0.01); (5)The values of alanine aminotransferase (ALT) and lactate dehydroge nase (LDH) in coculture supernatants of cagA (+) Hp and HepG2 were increased (P < 0.01); (6) No influence of cagA () Hp on HepG2 was found. CONCLUSIONS: cagA (+) Hp has in vitro cytotoxicity on HepG2 cells, and the cytotoxicity is positi vely related to the concentration and culture time of cagA(+) HP.

关 键 词：幽门螺杆菌 肝癌 HEPG2 体外细胞毒作用 癌细胞 

分 类 号：R735.7[医药卫生—肿瘤]