PDF(Pubmed)
Abstract：
OBJECTIVE: Glutathione peroxidases (GPXs) are crucial antioxidant enzymes, counteracting reactive oxygen species (ROS). GPX overexpression promotes proliferation and invasion in cancer cells. Glutathione peroxidase-1 (GPX1), the most abundant isoform, contributes to invasion, migration, cisplatin resistance, and proliferation in various cancers. Nuclear factor-kappa B (NF-[Formula: see text]B) participates in cell proliferation, apoptosis, and tumor progression. The inhibition of NF-[Formula: see text]B expression reduces the malignancy of esophageal squamous cell carcinoma. This study aimed to explore the GPX1 and NF-[Formula: see text]B signaling pathways and their correlation with gastric cancer cell proliferation and invasion.
METHODS: Cell culture, complementary DNA microarray analysis, western blotting, reverse transcription-polymerase chain reaction, zymography, 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay, GPX1 knock-down with short hairpin RNA (shRNA), standard two-chamber invasion assay, chromatin immunoprecipitation assay.
RESULTS: Hepatocyte growth factor (HGF) up-regulated GPX1 expression in gastric cancer cells. The NF-[Formula: see text]B inhibitor, pyrrolidine dithiocarbamate down-regulated HGF-induced GPX1 protein levels. Furthermore, NF-[Formula: see text]B and urokinase-type plasminogen activators were down-regulated in GPX1-shRNA-treated cells. Treatment with an Akt pathway inhibitor (LY294002) led to the down-regulation of GPX1 and NF-[Formula: see text]B gastric cancer cells. GPX1 knockdown resulted in decreased HGF-mediated in vitro cell proliferation and invasion. The study identified the putative binding site of the GPX1 promoter containing the NF-[Formula: see text]B binding site, confirmed through chromatin immunoprecipitation.
CONCLUSIONS: HGF induced GPX1 expression through the NF-[Formula: see text]B and Akt pathways, suggesting a central role in gastric cell proliferation and invasion. Hence, GPX1 emerges as a potential therapeutic target for gastric cancer.
METHODS: Cell culture, complementary DNA microarray analysis, western blotting, reverse transcription-polymerase chain reaction, zymography, 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay, GPX1 knock-down with short hairpin RNA (shRNA), standard two-chamber invasion assay, chromatin immunoprecipitation assay.
RESULTS: Hepatocyte growth factor (HGF) up-regulated GPX1 expression in gastric cancer cells. The NF-[Formula: see text]B inhibitor, pyrrolidine dithiocarbamate down-regulated HGF-induced GPX1 protein levels. Furthermore, NF-[Formula: see text]B and urokinase-type plasminogen activators were down-regulated in GPX1-shRNA-treated cells. Treatment with an Akt pathway inhibitor (LY294002) led to the down-regulation of GPX1 and NF-[Formula: see text]B gastric cancer cells. GPX1 knockdown resulted in decreased HGF-mediated in vitro cell proliferation and invasion. The study identified the putative binding site of the GPX1 promoter containing the NF-[Formula: see text]B binding site, confirmed through chromatin immunoprecipitation.
CONCLUSIONS: HGF induced GPX1 expression through the NF-[Formula: see text]B and Akt pathways, suggesting a central role in gastric cell proliferation and invasion. Hence, GPX1 emerges as a potential therapeutic target for gastric cancer.
摘要：
目的：谷胱甘肽过氧化物酶（GPXs）是重要的抗氧化酶，抵消活性氧（ROS）。GPX过表达促进癌细胞的增殖和侵袭。谷胱甘肽过氧化物酶-1(GPX1),最丰富的同种型，有助于入侵，迁移,顺铂耐药,和各种癌症的增殖。核因子-κB（NF-[公式：见正文]B）参与细胞增殖，凋亡，和肿瘤进展。NF-[公式:参见正文]B表达的抑制降低了食管鳞状细胞癌的恶性程度。本研究旨在探讨GPX1和NF-B信号通路及其与胃癌细胞增殖和侵袭的相关性。
方法：细胞培养，互补DNA微阵列分析，西方印迹,逆转录聚合酶链反应，酶谱,3-（4,5-二甲基噻唑-2-基）-2,5-二苯基四唑溴化物测定，用短发夹RNA（shRNA）敲低GPX1，标准的两室侵入试验，染色质免疫沉淀测定。
结果：肝细胞生长因子（HGF）上调胃癌细胞中GPX1的表达。NF-[配方：见正文]B抑制剂，吡咯烷二硫代氨基甲酸酯下调HGF诱导的GPX1蛋白水平。此外，NF-[式:参见正文]B和尿激酶型纤溶酶原激活剂在GPX1-shRNA处理的细胞中下调。用Akt途径抑制剂(LY294002)处理导致GPX1和NF-[配方:参见正文]B胃癌细胞的下调。GPX1敲低导致HGF介导的体外细胞增殖和侵袭减少。该研究确定了GPX1启动子的推定结合位点，其中含有NF-[公式：参见文本]B结合位点，通过染色质免疫沉淀证实。
结论：HGF通过NF-[公式：参见文本]B和Akt途径诱导GPX1表达，提示在胃细胞增殖和侵袭中起核心作用。因此，GPX1成为胃癌的潜在治疗靶点。
方法：细胞培养，互补DNA微阵列分析，西方印迹,逆转录聚合酶链反应，酶谱,3-（4,5-二甲基噻唑-2-基）-2,5-二苯基四唑溴化物测定，用短发夹RNA（shRNA）敲低GPX1，标准的两室侵入试验，染色质免疫沉淀测定。
结果：肝细胞生长因子（HGF）上调胃癌细胞中GPX1的表达。NF-[配方：见正文]B抑制剂，吡咯烷二硫代氨基甲酸酯下调HGF诱导的GPX1蛋白水平。此外，NF-[式:参见正文]B和尿激酶型纤溶酶原激活剂在GPX1-shRNA处理的细胞中下调。用Akt途径抑制剂(LY294002)处理导致GPX1和NF-[配方:参见正文]B胃癌细胞的下调。GPX1敲低导致HGF介导的体外细胞增殖和侵袭减少。该研究确定了GPX1启动子的推定结合位点，其中含有NF-[公式：参见文本]B结合位点，通过染色质免疫沉淀证实。
结论：HGF通过NF-[公式：参见文本]B和Akt途径诱导GPX1表达，提示在胃细胞增殖和侵袭中起核心作用。因此，GPX1成为胃癌的潜在治疗靶点。
