南方医科大学学报

南方医科大学学报 ›› 2024, Vol. 44 ›› Issue (2): 354-362.doi: 10.12122/j.issn.1673-4254.2024.02.18

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BZW1 高表达促进胃癌细胞的侵袭和转移：基于调控Wnt//β-catenin通路和促进上皮间质转化

张文静，张 诺，杨 子，张小凤，孙奥飞，王 炼，宋 雪，耿志军，李 静，胡建国   

  1. 蚌埠医学院第一附属医院检验科，胃肠外科，中心实验室，安徽 蚌埠 233004；炎症相关性疾病基础与转化研究安徽省重点实验室，安徽 蚌埠 233004
  • 发布日期:2024-03-14

Overexpression of BZW1 promotes invasion and metastasis of gastric cancer cells by regulating Wnt/β-catenin signaling and promoting epithelial-mesenchymal transition

ZHANG Wenjing, ZHANG Nuo, YANG Zi, ZHANG Xiaofeng, SUN Aofei, WANG Lian, SONG Xue, GENG Zhijun, LI Jing, HU Jianguo   

  1. Clinical Laboratory, Department of Gastrointestinal Surgery, Central Laboratory, First Affiliated Hospital of Bengbu Medical Collage, Bengbu 233004, China; Anhui Provincial Key Laboratory for Basic and Translational Research of Inflammation-Associated Diseases, Bengbu 233004, China
  • Published:2024-03-14

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摘要： 目的 明确碱性亮氨酸拉链蛋白1（BZW1）在胃癌中的表达情况，并探讨其对胃癌患者预后的影响及其可能的作用机制。方法 分别采用TIMER、UALCAN和Kaplan-Meier Plotter数据库分析BZW1在胃癌组织中的表达情况，与肿瘤分级、分期之间的相关性及对患者预后的影响。纳入2014年1月~2016年12月在我院行胃癌根治术的102例患者，分析BZW1在胃癌组织中的表达水平、对胃癌疾病进展和患者术后5年生存率的影响。体外采用慢病毒转染的方式构建上调和下调BZW1表达的胃癌细胞系（MGC803），分析BZW1对MGC803细胞迁移、侵袭及上皮-间质转化（EMT）的作用。采用KEGG富集分析预测BZW1在胃癌中的可能作用机制，并进一步采用Western blot实验进行体外验证。结果 生物信息学分析结果显示，BZW1在胃癌和癌旁组织中的表达量差异有统计学意义（P<0.01），免疫组化和RT-qPCR结果显示，BZW1在胃癌组织中的蛋白和mRNA表达水平分别是癌旁组织的3.30倍和6.54倍（P<0.01）；胃癌组织中BZW1的表达水平与外周血CEA和CA199水平呈正相关（P<0.01）；单因素结合Cox多元回归模型分析证实BZW1高表达（P<0.05，HR=2.070，95% CI：1.021~4.196）是影响胃癌患者根治术后5年生存率的独立危险因素；以BZW1相对表达量3.61为截点值，预判术后5年死亡的敏感性为75.56%，特异性为71.93%（P<0.01）。Transwell 实验显示，上调 BZW1 可促进 MGC803 细胞的迁移和侵袭（P<0.05），下调则相反（P<0.05）。Western blot实验发现，上调BZW1可促进MGC803细胞N-cadherin与Vimentin的表达（P<0.05），并抑制E-cadherin的表达（P<0.05），下调则反之（P<0.05）。富集分析显示，BZW1生物功能可能与Wnt//β-catenin信号相关。Western blot实验进一步证实，上调BZW1可促进Wnt3a、β-Catenin及C-myc的表达（P<0.05），而下调则相反（P<0.05）；使用Wnt通路抑制剂XAV-939可显著削弱上调BZW1对MGC803细胞中EMT关键分子的蛋白N-cadherin、Vimentin及E-cadherin的调控作用（P<0.05）。结论 BZW1在胃癌组织中高表达并影响患者预后，可能与调控Wnt/β-catenin信号促进胃癌细胞EMT进程相关。

关键词: 胃癌；碱性亮氨酸拉链蛋白1；预后；上皮-间质转化；Wnt/β-catenin

Abstract: Objective To investigate the expression level of basic leucine zipper and W2 domain-containing protein 1 (BZW1) in gastric cancer, its impact on patient prognosis and the underlying mechanisms. Methods TIMER, UALCAN and Kaplan-Meier Plotter databases were used for analyzing BZW1 expression level gastric cancer tissues and its correlation with tumor grade and stage and the patients' prognosis. We further analyzed BZW1 expressions, disease progression, and postoperative 5-year survival in 102 patients undergoing radical surgery for gastric cancer at our hospital between January, 2014 and December, 2016. Gastric cancer MGC803 cells were examined for changes in migration, invasion, and epithelial-mesenchymal transition (EMT) following lentivirus-mediated BZW1 overexpression or knockdown. Results The protein and mRNA expressions of BZW1 in gastric cancer tissues were 3.30 and 6.54 times of those in adjacent tissues, respectively (P<0.01). BZW1 expression in gastric cancer tissues were positively correlated with peripheral blood CEA and CA199 levels (P<0.01). A high BZW1 expression was an independent risk factor for 5-year survival of gastric cancer patients after radical surgery (P<0.05, HR=2.070, 95%CI: 1.021-4.196). At the cut-off value of 3.61, BZW1 expression had a sensitivity of 75.56% and a specificity of 71.93% for predicting postoperative 5-year mortality (P<0.01). In MGC803 cells, BZW1 overexpression obviously promoted cell migration and invasion (P<0.05), enhanced cellular expressions of N-cadherin and vimentin (P<0.05) and inhibited the expression of E-cadherin (P<0.05). Enrichment analysis suggested the involvement of BZW1 in the Wnt/β-catenin signaling pathway. Western blotting confirmed that BZW1 overexpression promoted while BZW1 knockdown inhibited the expressions of Wnt3a, β-catenin and C-myc in MGC803 cells (P<0.05). Conclusion BZW1 is highly expressed in gastric cancer tissues to affect the patient prognosis possibly by activation the Wnt/β-catenin signaling pathway to promote EMT of gastric cancer cells.

Key words: gastric cancer; basic leucine zipper and W2 domain-containing protein 1; prognosis; epithelial-mesenchymal transition; Wnt/β-catenin