南方医科大学学报

南方医科大学学报 ›› 2023, Vol. 43 ›› Issue (7): 1063-1070.doi: 10.12122/j.issn.1673-4254.2023.07.02

• • 上一篇    下一篇

高表达DAP5促进胃癌细胞的糖代谢并与不良预后相关

王秋生，张 震，王 炼，汪 煜，姚新宇，王月月，张小凤，葛思堂，左芦根   

  1. 蚌埠医学院第一附属医院胃肠外科，检验科，中心实验室，安徽 蚌埠 233004；蚌埠医学院，安徽 蚌埠 233030；蚌埠医学院组织移植重点实验室，安徽 蚌埠 233030
  • 出版日期:2023-07-20 发布日期:2023-07-20

High expression of death-associated protein 5 promotes glucose metabolism in gastric cancer cells and correlates with poor survival outcomes

WANG Qiusheng, ZHANG Zhen, WANG Lian, WANG Yu, YAO Xinyu, WANG Yueyue, ZHANG Xiaofeng, GE Sitang, ZUO Lugen   

  1. Department of Gastrointestinal Surgery, Clinical Laboratory, Central Laboratory, First Affiliated Hospital of Bengbu Medical College, Bengbu 233000, China; Bengbu Medical College, Bengbu 233030, China; Key Laboratory of Tissue Transplantation, Bengbu Medical College, Bengbu 233000, China
  • Online:2023-07-20 Published:2023-07-20

RichHTML

31

PDF

490

摘要： 目的 探讨死亡相关蛋白5（DAP5）在胃癌组织中表达的预后价值及其对癌细胞有氧糖酵解的作用。方法 采用公共数据库分析DAP5在胃癌及癌旁组织的表达及与患者预后的关系，纳入分析2012年6月~2017年7月于我院行胃癌根治术的患者102例，采用免疫组织化学法检测DAP5在胃癌和癌旁组织中的表达情况；统计学分析DAP5与临床病理参数间的关系；K-M生存分析评估DAP5对胃癌患者术后5年生存率的影响；Cox回归评估影响患者术后5年生存率的独立危险因素；ROC曲线分析DAP5表达对患者术后5年生存率的预测价值；基因富集分析预测DAP5在胃癌中的功能；采用慢病毒调控胃癌细胞系（MGC-803）DAP5的表达，将实验分为空白对照组、DAP5敲低组和DAP5过表达组；葡萄糖摄取实验、细胞乳酸水平测定、实时荧光定量PCR及免疫印迹实验分析DAP5对胃癌细胞有氧糖酵解的作用。结果 公共数据库分析显示DAP5在胃癌中高表达且与肿瘤进展及不良预后相关（P<0.05）；免疫组化结果显示，胃癌组织中 DAP5 表达水平较癌旁组织显著增高（3.19±0.60 vs 1.00±0.12，P<0.01）；K-M 生存分析显示 DAP5 高表达组患者术后 5 年生存率较 DAP5 低表达组明显降低（17.6%，9/51 vs 72.5%，37/51，P<0.05）；Cox回归分析进一步显示DAP5高表达、T3-4期、N2-3期、外周血CEA≥5 ng/mL是影响胃癌患者根治术后5年生存率的独立危险因素（P<0.05）；ROC曲线分析表明DAP5评估胃癌患者术后5年生存率的敏感性为73.2%，特异性为80.4%，准确度为79.0%（P<0.01）；体外实验显示，敲低DAP5可显著降低MGC-803细胞对葡萄糖摄取、乳酸水平及有氧糖酵解关键酶GLUT1、HK2、LDHA的mRNA和蛋白水平，过表达实验则相反（P<0.05）。结论 DAP5在胃癌中高表达并与患者不良预后相关，其可能通过促进癌细胞有氧糖酵解进程影响癌症进展，有望成为评估胃癌患者远期预后的潜在生物学标志物。

关键词: 胃癌；死亡相关蛋白5；生物信息学；预后；有氧糖酵解

Abstract: Objective To investigate the prognostic value of death-associated protein 5 (DAP5) in gastric cancer (GC) and its regulatory effect on aerobic glycolysis in GC cells. Methods We analyzed DAP5 expression levels in GC and adjacent tissues and its association with survival outcomes of GC patients using public databases. We collected paired samples of GC and djacent tissues from 102 patients undergoing radical resection of GC in our hospital from June, 2012 to July, 2017, and analyzed the correlation of DAP5 expression level detected immunohistochemically with the clinicopathological parameters of the patients. Cox regression analysis, Kaplan-Meier analysis, and ROC curves were used to explore the independent risk factors and the predictive value of DAP5 expression for 5-year survival of the patients. In the cell experiments, we observed the changes in aerobic glycolysis in MGC-803 cells following lentivirus-mediated DAP5 knockdown or overexpression by measuring glucose uptake and cellular lactate level and using qRT-PCR and Western blotting. Results Analysis using the public databases showed that DAP5 was highly expressed in GC and correlated with tumor progression and poor survival outcomes of the patients (P<0.05). In the clinical samples, DAP5 expression was significantly higher in GC than in the adjacent tissues (3.19±0.60 vs 1.00±0.12; t=36.863, P<0.01), and a high expression of DAP5 was associated with a reduced 5-year survival rate of the patients (17.6% vs 72.5%; χ2=29.921, P<0.05). A high DAP5 expression, T3-4, N2-3, and CEA≥5 ng/mL were identified as independent risk factors affecting 5-year survival outcomes of GC (P<0.05), for which DAP5 expression showed a prediction sensitivity, specificity and accuracy of 73.2%, 80.4% and 79.0%, respectively. In MGC-803 cells, DAP5 knockdown significantly reduced glucose uptake, lactate level and the expressions of GLUT1, HK2 and LDHA, and DAP5 overexpression produced the opposite effects (P<0.05). Conclusion A high expression of DAP5 in GC, which enhances cellular aerobic glycolysis to promote cancer progression, is correlated with a poor survival outcome and may serve as a biomarker for evaluating long-term prognosis of GC patients.

Key words: gastric cancer; death-associated protein 5; bioinformatics; prognosis; aerobic glycolysis