南方医科大学学报

南方医科大学学报 ›› 2023, Vol. 43 ›› Issue (10): 1715-1724.doi: 10.12122/j.issn.1673-4254.2023.10.09

• • 上一篇    下一篇

ANKRD6高表达是结肠癌不良预后的有效预测指标

赵海远，刘 刚，李 阳，杨年钊，赵 军   

  1. 皖南医学院弋矶山医院//皖南医学院第一附属医院胃肠外科，安徽 芜湖 241001
  • 出版日期:2023-10-20 发布日期:2023-11-03

High expression of ANKRD6 is an indicator for poor prognosis of colon adenocarcinoma

ZHAO Haiyuan, LIU Gang, LI Yang, YANG Nianzhao, ZHAO Jun   

  1. Department of General Surgery, Yijishan Hospital of Wannan Medical College/First Affiliated Hospital of Wannan Medical College, Wuhu 241001, China
  • Online:2023-10-20 Published:2023-11-03

RichHTML

18

PDF

330

摘要： 目的 探讨ANKRD6基因在结肠癌（COAD）中的表达情况以及其在结肠癌预后和治疗中的潜在作用。方法 通过免疫组化检测20例配对的结直肠腺癌组织和癌旁组织，利用实时定量PCR方法比较结肠癌细胞系和正常结肠细胞中ANKRD6基因的表达差异。通过TCGA数据库下载521例COAD患者和41例正常组织的基因表达矩阵、免疫浸润数据和临床信息，利用R软件分析ANKRD6在COAD组织中与临床病理特征的表达差异、相关基因集富集和免疫浸润情况。采用单因素和多因素Cox回归以及 Kaplan-Meier 方法评估 COAD 的预后及相关因素，并构建含 ANKRD6 的预后列线图。最后进行体外实验，通过Western blot、CCK-8实验、划痕实验和Transwell检测siANKRD6与siCtrl的COAD细胞生物学活性。结果 ANKRD6在COAD细胞系中高表达，并在恶性程度较高的COAD组织中表达上调（P<0.05）。ANKRD6的表达上调与COAD患者的不良预后（总体生存期和无病生存期）相关（P<0.05）。单因素和多因素Cox回归分析显示，高表达的ANKRD6与患者整体生存率相关（P<0.05）。在此基础上，构建了预后Nomogram，其C-index值为0.739，能有效地预测患者1年、3年和5年的生存预后因素，Calibration校准曲线、DCA曲线和ROC曲线评估该模型的准确性较高。免疫浸润分析结果显示，ANKRD6的表达与巨噬细胞等免疫浸润程度呈正相关（P<0.05），而与Th17细胞等免疫浸润程度呈负相关（P<0.05）。GOKEGG的富集分析结果显示，ANKRD6在COAD中主要富集于Wnt信号通路、血管内皮生长因子受体信号通路和钙信号通路等。体外实验显示，相对于siCtrl，siANKRD6可减弱COAD细胞的增殖、迁移和侵袭能力（P<0.05）。结论 ANKRD6是COAD预后不良的有效预测指标，为COAD的诊断、治疗靶点的寻找和个体化治疗等提供了参考。

关键词: 结肠癌；ANKRD6；肿瘤免疫浸润；预后

Abstract: Objective To explore the expression of ANKRD6 in colon adenocarcinoma (COAD) and its implications for prognosis and treatment of COAD. Methods We investigated the differential expression of ANKRD6 in 20 pairs of COAD and adjacent tissues using immunohistochemistry and in colon cancer cell lines and normal colon cells using real-time quantitative PCR. Gene expression matrices, immune infiltration data, and clinical information of 521 COAD patients and 41 normal tissues were obtained from the TCGA database for analyzing the association of ANKRD6 expression with clinicopathological features, gene set enrichment, and immune infiltration in COAD using R software. The prognostic factors of COAD were evaluated using univariate and multivariate Cox regression and Kaplan-Meier analyses, and a prognostic nomogram containing ANKRD6 was constructed. Western blotting, CCK-8 assay, scratch assay and Transwell assay were performed to assess the effects of ANKRD6 knockdown on biological activities of COAD cells. Results ANKRD6 was highly expressed in COAD cell lines and upregulated in COAD tissues with higher malignancy (P<0.05). Increased ANKRD6 expression was significantly correlated with decreased overall survival and disease-free survival of COAD patients (P<0.05). Univariate and multivariate Cox regression analyses indicated that ANKRD6 expression was significantly correlated with the overall survival rate of COAD patients (P<0.05). The constructed prognostic nomogram showed a C-index value of 0.739 and effectively predicted the 1- , 3- , and 5-year survival of the patients. ANKRD6 expression was positively correlated with immune infiltration of macrophages (P<0.05) and negatively correlated with Th17 cells and other immune cell infiltration (P<0.05). Bioinformatic analysis suggested that ANKRD6 was mainly enriched in Wnt, vascular endothelial growth factor receptor, and calcium signaling pathways in COAD. In cultured COAD cells, ANKRD6 knockdown significantly suppressed cell proliferation, migration, and invasion (P<0.05). Conclusion High ANKRD6 expression is an indicator for poor prognosis of COAD, and ANKRD6 may serve as a potential biomarker as well as a therapeutic target for diagnosis and treatment of COAD.

Key words: colon adenocarcinoma; ANKRD6; tumor immune infiltration; prognosis