DBNDD1基因在结直肠癌进展过程中的作用与机制研究

doi:10.19586/j.2095-2341.2025.0033

生物技术进展 ›› 2025, Vol. 15 ›› Issue (4): 726-734.DOI: 10.19586/j.2095-2341.2025.0033

• 研究论文 • 上一篇

DBNDD1基因在结直肠癌进展过程中的作用与机制研究

刘晓雅¹^,²(), 张硕珉², 郑鹏², 马睿², 张朝军¹^,²()

^1.华南理工大学医学院，广州 510006
^2.解放军总医院第一医学中心普通外科，北京 100853

收稿日期:2025-03-11 接受日期:2025-05-06 出版日期:2025-07-25 发布日期:2025-09-08
通讯作者: 张朝军
作者简介:刘晓雅 E-mail: Lxy_208@163.com；
基金资助:
国家自然科学基金项目(81972320)

Role and Mechanisms of DBNDD1 in Colorectal Cancer Development

Xiaoya LIU¹^,²(), Shuomin ZHANG², Peng ZHENG², Rui MA², Chaojun ZHANG¹^,²()

^1.School of Medicine，South China University of Technology，Guangzhou 510006，China
^2.Department of General Surgery，First Medical Center，People's Liberation Army General Hospital，Beijing 100853，China

Received:2025-03-11 Accepted:2025-05-06 Online:2025-07-25 Published:2025-09-08
Contact: Chaojun ZHANG

摘要/Abstract

摘要：

结直肠癌（colorectal cancer，CRC）的恶性进展与关键癌基因的异常激活密切相关。基于癌症基因组图谱（the cancer genome atlas，TCGA）组学数据库分析结果显示，DBNDD1基因在CRC组织中高表达，且与患者总生存期缩短存在显著相关性。探究了DBNDD1在CRC中的表达特征、生物学功能及临床意义。通过TCGA数据库评估DBNDD1表达与预后的关联性；利用siRNA敲低DBNDD1后检测HCT116细胞的迁移、侵袭能力（划痕实验、Transwell实验）及凋亡水平（流式细胞术）；构建裸鼠移植瘤模型观察肿瘤生长及凋亡变化（Ki-67免疫组化、TUNEL检测），并结合基因集富集分析（gene set enrichment analysis，GESA）探索相关信号通路及免疫微环境调控机制。实验结果表明，DBNDD1敲低可抑制HCT116细胞的迁移及侵袭能力，同时促进细胞凋亡。进一步的机制分析显示，DBNDD1高表达样本在NF-κB和TNF信号通路上呈现显著富集，并与免疫抑制微环境特征（如M2型巨噬细胞及调节性T细胞浸润增加）存在密切关联。研究结果可为结直肠癌的临床分子标志物开发及靶向干预策略提供参考。

关键词: 结直肠癌, DBNDD1, 肿瘤微环境, NF-κB信号通路, 免疫浸润

Abstract:

The malignant progression of colorectal cancer （CRC） is closely associated with abnormal activation of critical oncogenes. Analysis of the cancer genome atlas （TCGA） omics data revealed that DBNDD1 is significantly upregulated in CRC tissues， with its overexpression demonstrating a significant correlation with shorter overall patient survival. This study aimed to investigate the expression characteristics， biological functions， and clinical significance of DBNDD1 in CRC. The research integrated omics analysis with experimental validation， evaluating the prognostic relevance of DBNDD1 expression through the TCGA database， assessing migration， invasion capabilities （Transwell assay， wound healing assay）， and apoptosis levels （flow cytometry） in HCT116 cells following DBNDD1 knockdown using siRNA， and monitoring tumor growth and apoptosis changes （Ki-67 immunohistochemistry， TUNEL assay） in nude mouse xenograft models. Gene set enrichment analysis （GSEA） was further employed to explore associated signaling pathways and immune microenvironment regulatory mechanisms. The experimental results demonstrated that DBNDD1 knockdown suppressed migration and invasion capabilities of HCT116 cells while promoting apoptosis. Mechanistic analysis revealed significant enrichment of NF-κB and TNF signaling pathways in DBNDD1-overexpressing samples， along with close associations with immunosuppressive microenvironment features， including increased infiltration of M2 macrophages and regulatory T cells. These findings provide novel research directions for the development of clinical molecular biomarkers and targeted therapeutic strategies in colorectal cancer.

Key words: colorectal cancer, DBNDD1, tumor microenvironment, NF-κB signaling pathway, immune infiltration

中图分类号:

Q291

刘晓雅, 张硕珉, 郑鹏, 马睿, 张朝军. DBNDD1基因在结直肠癌进展过程中的作用与机制研究[J]. 生物技术进展, 2025, 15(4): 726-734.

Xiaoya LIU, Shuomin ZHANG, Peng ZHENG, Rui MA, Chaojun ZHANG. Role and Mechanisms of DBNDD1 in Colorectal Cancer Development[J]. Current Biotechnology, 2025, 15(4): 726-734.

图/表 7

表1 本研究所用引物序列

Table 1 Primer sequences used in this study

引物名称

引物序列（5′→3′）

长度/bp

DBNDD1

F：5'-GGACCTGAAGATGGTGAAGG-3'

R：5'-CAGCACAGTCATCACCACAT-3'

150

GAPDH

F：5'-TGCACCACCAACTGCTTAG-3'

R：5'-GGCATGGACTGTGGTCATGAG-3'

130

图1 DBNDD1在泛癌和结直肠癌中的表达A：DBNDD1基因在不同类型癌症中的表达分析。ACC—肾上腺皮质癌；BLCA—膀胱尿路上皮癌；BRCA—乳腺癌；CESC—宫颈鳞状细胞癌；CHOL—胆管癌；COAD—结直肠癌；DLBC—弥漫性大B细胞淋巴瘤；ESCA—食管癌；GBM—胶质母细胞瘤；HNSC—头颈部鳞状细胞癌；KICH—肾嫌色细胞癌；KIRC—肾透明细胞癌；KIRP—肾乳头状细胞癌；LAML—急性髓系白血病；LGG—低级别神经胶质瘤；LIHC—肝细胞癌；LUAD—肺腺癌；LUSC—肺鳞状细胞癌；MESO—间皮瘤；OV—卵巢浆液性癌；PAAD—胰腺癌；PCPG—嗜铬细胞瘤和副神经节瘤；PRAD—前列腺腺癌；READ—直肠腺癌；SARC—肉瘤；SKCM—皮肤黑色素瘤；STAD—胃癌；TGCT—睾丸生殖细胞肿瘤；THCA—甲状腺癌；THYM—胸腺癌；UCEC—子宫内膜癌；UCS—子宫肉瘤；UVM—葡萄膜黑色素瘤；T—肿瘤组织；N—正常组织。B：DBNDD1在正常组织与肿瘤组织中的表达箱线图。C：依据DBNDD1表达水平（高/低）分层的Kaplan-Meier生存曲线；红色癌症缩写名称代表DBNDD1在该癌种中表达上调，绿色癌症缩写名称代表DBNDD1在该癌种中表达下调，黑色癌症缩写名称代表DBNDD1在该癌种的表达与正常组织无明显差异。

Fig. 1 Expression of DBNDD1 in pan-cancer and colorectal cancers

图2 体外验证DBNDD1在肠上皮及癌细胞中的表达差异A：DBNDD1在不同细胞系中的表达特征；B：siRNA干扰对DBNDD1表达的影响；每组n=3

Fig. 2 In vitro validation of differential DBNDD1 expression between intestinal epithelial and cancer cells

图3 DBNDD1基因敲低抑制结直肠癌细胞的恶性生物学行为A~B：敲低DBNDD1基因对结肠癌细胞迁移能力的影响（细胞划痕实验）以及划痕愈合率定量分析（比例尺=200 μm）；C~D：敲低DBNDD1基因对细胞侵袭能力的抑制作用（Transwell基质胶侵袭实验）以及侵袭细胞数定量统计（比例尺=50 μm）；E：细胞凋亡分析；每组n=3；*、**、***分别表示在P<0.05、P<0.01和P<0.001水平上具有统计学意义。

Fig. 3 DBNDD1 knockdown suppresses malignant biological behaviors in colorectal cancer cells

图4 体内实验证实DBNDD1驱动结直肠癌恶性进展A：来自Si-NC和Si-DBNDD1处理组的肿瘤图像；B：肿瘤体积的定量分析；C：Ki-67免疫组织化学染色（比例尺=20 μm）；D：TUNEL染色检测凋亡（比例尺=20 μm）；E：Ki-67表达的定量分析；F：总凋亡率的定量分析。每组n=4。

Fig. 4 DBNDD1 drives metastatic dissemination of colorectal cancer in vivo

图5 DBNDD1相关基因GO富集结果和KEGG通路解析

Fig. 5 GO functional annotation and KEGG pathway enrichment analysis of DBNDD1-associated genes

图6 免疫细胞浸润热图分析

Fig. 6 Heatmap analysis of immune cell infiltration

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DBNDD1基因在结直肠癌进展过程中的作用与机制研究

Role and Mechanisms of DBNDD1 in Colorectal Cancer Development

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