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利用聚(ADP-核糖)聚合酶(PARP)裂解评估不同幽门螺杆菌菌株对胃腺癌细胞系和成纤维细胞系凋亡的诱导作用。

Evaluation of apoptosis induction using PARP cleavage on gastric adenocarcinoma and fibroblast cell lines by different strains of Helicobacter pylori.

作者信息

Mojtahedi Ali, Salehi Rasoul, Navabakbar Farahtaj, Tamizifar Hasan, Tavakkoli Hamid, Duronio Vincent

机构信息

Department of Medical Microbiology, Faculty of Medicine, Guilan University Complex, Rasht-Tehran Road, Rasht 3477, Iran.

出版信息

Pak J Biol Sci. 2007 Nov 15;10(22):4097-102. doi: 10.3923/pjbs.2007.4097.4102.

Abstract

Helicobacter pylori is one of the most common pathogens affecting humans and is the major environmental factor in the development of gastric cancer increasing from 4 to 6 folds the risk of its development. Variations in cancer risk among H. pylori infected individuals may correlate to difference in H. pylori strains, variable host characteristics and specific interactions between host and microbial determinants. To determine the effect of different strains of H. pylori on cellular apoptosis this study was designed an in vitro model using AGS and HEF cell lines. After specified time intervals total cell proteins was extracted and subjected to SDS-PAGE and immunoblotting using anti poly ADP-ribose polymerase (PARP) antibody. Decrease in densitometric value of PARP was indicative of higher level of apoptosis. The ability of apoptosis induction in AGS and HEF cell lines by wild type (cagA+/vacA+), cagA-/vacA+, vacA-/cagA+ and double negative (cagA-/vacA-) strains were significantly different. The assessed apoptosis in AGS cell line co-cultured with wild type strain was 3.22 +/- 0.2 in 24 h, 2.8 +/- 0.1 in 48 and 2.1 +/- 0.09 in 72 h of incubation time. Similar assessment with cagA-/vacA+ strains in AGS cells was 4.17 +/- 1.49 in 24 h, 3.32 +/- 0.45 in 48 h and 2.32 +/- 0.61 in 72 h incubation. A variation in apoptotic potential between the H. pylori strains on two cells (AGS and HEF) was observed. Based on present results, it is concluded that H. pylori strains as well as target cell types are important in pathogenesis and induction of apoptosis during a specified time interval.

摘要

幽门螺杆菌是影响人类的最常见病原体之一,是胃癌发生的主要环境因素,会使胃癌发生风险增加4至6倍。幽门螺杆菌感染个体之间癌症风险的差异可能与幽门螺杆菌菌株的差异、宿主特征的变化以及宿主与微生物决定因素之间的特定相互作用有关。为了确定不同幽门螺杆菌菌株对细胞凋亡的影响,本研究设计了一种使用AGS和HEF细胞系的体外模型。在特定时间间隔后,提取总细胞蛋白,并使用抗聚ADP-核糖聚合酶(PARP)抗体进行SDS-PAGE和免疫印迹。PARP光密度值的降低表明凋亡水平较高。野生型(cagA+/vacA+)、cagA-/vacA+、vacA-/cagA+和双阴性(cagA-/vacA-)菌株在AGS和HEF细胞系中诱导凋亡的能力存在显著差异。与野生型菌株共培养的AGS细胞系在孵育24小时时评估的凋亡率为3.22±0.2,48小时时为2.8±0.1,72小时时为2.1±0.09。AGS细胞中与cagA-/vacA+菌株的类似评估在24小时孵育时为4.17±1.49,48小时时为3.32±0.45,72小时时为2.32±0.61。观察到两种细胞(AGS和HEF)上幽门螺杆菌菌株之间凋亡潜力的差异。根据目前的结果,得出结论:幽门螺杆菌菌株以及靶细胞类型在特定时间间隔内的发病机制和凋亡诱导中很重要。

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