Effects of Helicobacter pylori γ-Glutamyltranspeptidase on Apoptosis and Inflammation in Human Biliary Cells
Effects of Helicobacter pylori γ-Glutamyltranspeptidase on Apoptosis and Inflammation in Human Biliary Cells
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DOI：
10.1007/s10620-012-2216-2
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发表时间：
2012-10-01
影响因子：
3.1
通讯作者：
Baik, Seung-Chul
中科院分区：
文献类型：
--
作者：
Boonyanugomol, Wongwarut;Chomvarin, Chariya;Baik, Seung-Chul
几项研究报道了肝合疾病个体中幽门螺杆菌的存在，但仍需要在体外和体内研究。在这里，我们确定了幽门螺杆菌γ-谷氨基转肽酶（GGT）对人cholangiocarcinoma瘤细胞系（KKU-100细胞）中凋亡和IL-8产生的诱导的影响。细胞活力和BRDU合成。分别分析。进行了RT-PCR和Western印迹分析，分别评估基因和蛋白质表达。通过ELISA暴露于幽门螺杆菌GGT（+）菌株，在KKU-100细胞中分泌IL-8分泌，与暴露于幽门螺杆菌GGT突变体菌株的细胞相比，KKU-100细胞的存活率降低了KKU-100细胞的存活和DNA合成。重组幽门螺杆菌GGT（RHP-GGT）的治疗大大降低了细胞的存活和DNA合成，并刺激了凋亡。这些特征对应于用RHP-GGT处理的KKU-100细胞中iNOS基因表达水平升高。 RT-PCR和Western印迹分析表明，RHP-GGT治疗增强了促凋亡分子（Bax，Caspase-9和Caspase-3）的表达，并下调了抗凋亡分子的表达（Bcl-2和Bcl-2 -xl）。用RHP-GGT处理后，未表达外内介导的凋亡分子，包括Fas和活化的caspase-8。此外，RHP-GGT在KKU-100细胞中显着刺激了IL-8分泌。您的数据表明，幽门螺杆菌GGT可能通过改变细胞动力学并促进炎症而参与肝胆细胞中癌症的发展。
Several studies have reported the presence of H. pylori in individuals with hepatobiliary diseases, but in vitro and in vivo studies are still needed. Here, we determined the effects of H. pylori gamma-glutamyltranspeptidase (GGT) on the induction of apoptosis and IL-8 production in a human cholangiocarcinoma cell line (KKU-100 cells).Cell viability and DNA synthesis were examined by MTT and BrdU assays, respectively. RT-PCR and western blot analysis were performed to assess gene and protein expression, respectively. IL-8 secretion in KKU-100 cells was measured by ELISA.Exposure to the H. pylori ggt (+) strain decreased KKU-100 cell survival and DNA synthesis when compared with cells exposed to the H. pylori ggt mutant strain. Treatment with recombinant H. pylori GGT (rHP-GGT) dramatically decreased cell survival and DNA synthesis, and stimulated apoptosis; these features corresponded to an increased level of iNOS gene expression in KKU-100 cells treated with rHP-GGT. RT-PCR and western blot analyses revealed that rHP-GGT treatment enhanced the expression of pro-apoptotic molecules (Bax, Caspase-9, and Caspase-3) and down-regulated the expression of anti-apoptotic molecules (Bcl-2 and Bcl-xL). The extrinsic-mediated apoptosis molecules, including Fas and activated Caspase-8, were not expressed after treatment with rHP-GGT. Furthermore, rHP-GGT significantly stimulated IL-8 secretion in KKU-100 cells.Our data indicate that H. pylori GGT might be involved in the development of cancer in hepatobiliary cells by altering cell kinetics and promoting inflammation.