Cortactin Mediates Apoptosis of Gastric Epithelial Cells Induced by VacA Protein of Helicobacter pylori
Cortactin Mediates Apoptosis of Gastric Epithelial Cells Induced by VacA Protein of Helicobacter pylori
复制标题
Cortactin介导幽门螺杆菌VacA蛋白诱导的胃上皮细胞凋亡
DOI：
10.1007/s10620-015-3836-0
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发表时间：
2016-01-01
影响因子：
3.1
通讯作者：
Lan, Chunhui
中科院分区：
文献类型：
--
作者：
Chang, Hui;Chen, Dongfeng;Lan, Chunhui
幽门螺杆菌（Helicobacter pylori，H. pylori），其负责细胞空泡变性和凋亡性细胞死亡。介导这一过程的候选宿主因子是corneumen，一种与H.为探讨Corpylin在VacA诱导的胃上皮细胞凋亡中的作用，构建了Corpylin表达载体和shRNA慢病毒载体，并转染人胃癌细胞系AGS。从H. pylori培养物，酸活化，并与转导的细胞群共孵育。流式细胞仪检测细胞凋亡，Western blot检测促凋亡蛋白Bax和抗凋亡蛋白Bcl-2的表达。coronin（AGS/coronin）表达增加导致更大百分比的细胞发生凋亡。相反，用shRNA敲低cortactin（AGS/cortactin-shRNA）降低了凋亡细胞的百分比。促凋亡蛋白Bax和抗凋亡蛋白Bcl-2的蛋白水平在AGS/corneum细胞中相对于亲本AGS细胞增加和减少。在AGS/cortactin-shRNA细胞中，Bax蛋白表达减少，Bcl-2蛋白表达增加，表明cortactin参与了VacA诱导的胃细胞凋亡的调控。
Vacuolating cytotoxin antigen (VacA) is one of the major virulence factors in Helicobacter pylori (H. pylori), which is responsible for cell vacuolar degeneration and apoptotic cell death. A candidate host factor which mediates this process is cortactin, a protein associated with the processes of colonization and adhesion of H. pylori in gastric epithelium.To investigate the role of cortactin in VacA-induced apoptosis of gastric epithelial cells.Cortactin expression and shRNA lentiviral constructs were developed and transduced into the human gastric cancer cell line, AGS. VacA protein was purified from H. pylori cultures, acid-activated, and co-incubated with the transduced cell populations. Apoptosis was detected by flow cytometry, and the levels of the pro- and anti-apoptotic proteins Bax and Bcl-2 were determined by Western blot.Acid-activated purified VacA induced apoptosis in the parental AGS cells. Increased expression of cortactin (AGS/cortactin) led to a greater percentage of cells undergoing apoptosis. In contrast, knockdown of cortactin with shRNA (AGS/cortactin-shRNA) decreased the percentage of apoptotic cells. The protein levels of pro- and anti-apoptotic proteins Bax and Bcl-2 were increased and decreased in AGS/cortactin cells relative to the parental AGS cells. In the AGS/cortactin-shRNA cells, Bax protein levels were decreased, while Bcl-2 protein was increased.The results indicate that cortactin is involved in the regulation of apoptosis induced by VacA in gastric cells.