磷酸烯醇式丙酮酸羧激酶1对胃癌细胞生物学行为的影响及相关机制

目的： 探讨磷酸烯醇式丙酮酸羧激酶1(phosphoenolpyruvate carboxykinase 1，PCK1)在胃癌中的表达、与胃癌患者预后的关系及其潜在机制。方法： 利用TCGA数据库、HPA数据库及Oncomine 4.5数据库中胃癌以及癌旁组织的相关数据与图片，分析PCK1表达水平与胃癌患者临床病理学特征和总生存期的关系，采用基因功能富集分析(GSEA)预测PCK1调控胃癌潜在的信号通路。体外细胞学实验采用慢病毒转染胃癌HGC-27和AGS细胞，分别敲低与过表达PCK1，采用细胞功能学实验检测PCK1对胃癌细胞生长、增殖、迁移等影响，同时采用蛋白质免疫印迹检测相关蛋白的表达。结果： 与癌旁组织相比，PCK1在胃癌组织中表达明显上调(P＜0.05)；ROC曲线提示PCK1对胃癌患者预后具有良好的预测效能；通过GSEA富集分析提示原发性免疫缺陷、细胞黏附分子、趋化因子信号通路、全身性红斑狼疮、产生IgA的肠道免疫网络及RIGⅠ样受体信号通路相关的基因集在PCK1基因低表达表型中差异富集；敲低PCK1后，胃癌HGC-27和AGS细胞生长、克隆形成能力、迁移能力显著下降，同时p-AKT(Ser473)、p-mTOR、p-S6和p-4EBP1表达水平明显降低(P＜0.05或P＜0.01)；过表达PCK1则与之作用相反。结论： PCK1在胃癌中呈高表达且与患者预后相关，其可能通过活化AKT mTOR信号通路促进胃癌细胞生长、增殖与迁移。

Effect of phosphoenolpyruvate carboxykinase 1 on the biological behavior of gastric cancer cells and related mechanisms

 Objective To investigate the expression level of phosphoenolpyruvate carboxykinase 1(PCK1) in gastric cancer, its relationship with the prognosis of gastric cancer patients and its potential mechanism. Methods By using the relevant data and images of gastric cancer and adjacent tissues in the TCGA database, HPA database and Oncomine 4.5 database, the relationship between the expression level of PCK1 and the clinicopathological features and overall survival of patients with gastric cancer was analyzed, and the potential signaling pathway of PCK1 regulation of gastric cancer was predicted by gene function enrichment analysis (GSEA). In vitro cytology experiment, lentivirus was used to transfect gastric cancer HGC-27 and AGS cells to knock down or over express PCK1, respectively. The effects of PCK1 on the growth, proliferation and migration of gastric cancer cells were detected by cell functional assay. And Western blotting was used to detect the expression of related proteins. Results Compared with the adjacent tissues, PCK1 expression was significantly up regulated in gastric cancer tissues (P＜0.05); ROC curve suggested that PCK1 had a significant efficacy in predicting prognosis of gastric cancer patients; enrichment analysis by GSEA suggested that gene sets related to primary immunodeficiency, cell adhesion molecules, chemokine signalling pathway, systemic lupus erythematosus, IgA producing intestinal immune network and RIGⅠ type receptor signalling pathway were differentially enriched in PCK1 gene low expression phenotype. Knockdown of PCK1, the growth, clonogenic capacity, and migration ability of gastric cancer HGC-27 and AGS cells were greatly decreased, and the expression levels of p AKT (Ser473), p-mTOR, p-S6 and p-4EBP1 were also markedly decreased (P<0.05 or P＜0.01); overexpression of PCK1 produced the opposite effect. Conclusion PCK1 is highly expressed in gastric cancer and is associated with the prognosis of patients, which may promote the growth, proliferation and migration of gastric cancer cells by activating the AKT-mTOR signalling pathway.