ELFN1高表达是结肠癌的预后生物标志物并促进结肠癌细胞的增殖转移

doi:10.12122/j.issn.1673-4254.2025.07.22

摘要/Abstract

摘要：

目的探讨细胞外富含亮氨酸重复序列和纤连蛋白III型结构域的蛋白质1（ELFN1）在结肠癌中的表达、临床预后、基因功能、信号通路和生物学表型。方法运用R语言，通过公共数据库TCGA、GEO数据库分析33种癌谱ELFN1表达水平，并鉴定结肠癌中ELFN1的差异基因，通过基因功能注释和富集分析鉴定其相关信号通路；分析并评价ELFN1表达与临床病理特征的相关性；qPCR及Western blotting验证ELFN1在细胞株以及肠癌组织中的表达水平；Transwell、EDU实验分析ELFN1在SW480细胞的生物学表型。结果 ELFN1在14种肿瘤中高表达，结肠癌中ELFN1的表达高于癌周组织（P<0.001）；临床特征与预后分析结果显示，ELFN1 mRNA高表达的患者OS较差；Cox回归分析显示，ELFN1表达是OS的独立预后因素（P<0.05）；ELFN1在肿瘤转移增殖中显著富集，进一步GSEA分析识别了与之相关的肿瘤信号通路；实验分析结果显示，肠癌细胞系ELFN1的表达水平高于正常细胞（P<0.05）；细胞功能实验显示，敲低ELFN1可阻止肠癌SW480细胞增殖和转移（P<0.05）。结论 ELFN1在结肠癌中异常高表达，并代表了不良的临床预后；并与肠癌增殖转移表型相关，提示ELFN1可能为结肠癌的预后标记物。

关键词: 结肠癌, ELFN1, ECM信号通路, 临床预后, 生物信息学

Abstract:

Objective To explore the correlation of ELFN1 expression level with prognosis of colorectal cancer and its regulatory role in colorectal cancer cell proliferation and metastasis. Methods We analyzed the expression levels of ELFN1 across 33 cancer types using publicly available databases and identified differential genes related to ELFN1 in colorectal cancer. Gene function annotation and enrichment analysis were used to identify the involved signaling pathways. Logistic analysis, Kaplan-Meier analysis and Cox regression analysis were performed to evaluate the correlation between ELFN1 expression and clinicopathological parameters and survival of colorectal cancer patients. qPCR and Western blotting were used to validate the expression levels of ELFN1 in different colorectal cancer cell lines and tissues, and Transwell and EDU experiments were carried out to assess the effect of ELFN1 knockdown on biological behaviors of SW480 cells. Results ELFN1 was highly expressed in 14 cancers, and its expression was significantly higher in colon cancer tissues than in adjacent tissues. A high expression of ELFN1 mRNA was associated with a poorer overall survival of colorectal cancer patients. Cox regression analysis indicated that ELFN1 expression was an independent prognostic factor for overall survival of the patients. ELFN1 was significantly enriched in tumor metastasis and proliferation and participated in several tumor signaling pathways. The colon cancer cell lines showed significantly higher expression levels of ELFN1 than normal cells, ELFN1 knockdown obviously inhibited proliferation and migration of SW480 cells in vitro. Conclusion ELFN1 is overexpressed in colorectal cancer and is associated with poor clinical prognosis of the patients. A high ELFN1 expression is associated with malignant phenotypes of colorectal cancer and promotes cancer cell proliferation and metastasis, suggesting its potential as a prognostic biomarker for colorectal cancer.

Key words: colorectal cancer, ELFN1, ECM signaling pathway, clinical prognosis, bioinformatics

王康, 李海宾, 余靖, 孟源, 张虹丽. ELFN1高表达是结肠癌的预后生物标志物并促进结肠癌细胞的增殖转移[J]. 南方医科大学学报, 2025, 45(7): 1543-1553.

Kang WANG, Haibin LI, Jing YU, Yuan MENG, Hongli ZHANG. High expression of ELFN1 is a prognostic biomarker and promotes proliferation and metastasis of colorectal cancer cells[J]. Journal of Southern Medical University, 2025, 45(7): 1543-1553.

图/表 9

图1 基于TCGA数据库显示ELFN1在多种肿瘤中高表达并具有预后差异

Fig.1 Data from the TCGA database show high expression of ELFN1 in a variety of tumors, which has different prognostic implications. A: TCGA diffuse carcinoma analysis. B: TCGA pan-cancer pairing analysis. C: Forest map of the prognostic value of ELFN1 in different tumors. *P<0.05, **P<0.01, ***P<0.001.

图2 ELFN1在结肠癌中具有较高诊断价值并在结肠癌中高表达

Fig.2 ELFN1 has a high diagnostic value in colon cancer and is highly expressed in colon cancer. A: Venn diagram of ELFN1 expression patterns and its prognosis and diagnostic value of colon cancer. B: ROC curve analysis showing an AUC value of ELFN1 in COAD of 0.851. C: High expression levels of ELFN1 in paired samples of TCGA. D-F: Multiple datasets in the GEO database indicate that the expression level of ELFN1 is higher tumor tissues than in normal tissues. *P<0.05, **P<0.01, ***P<0.001.

图3 ELFN1在结肠癌中的预后及临床价值

Fig.3 Prognostic and clinical significance of ELFN1 in colon cancer. A-C: Kaplan-Meier survival analysis reveals that COAD patients with high ELFN1 expression have shorter overall survival (A), disease-specific survival (B) and progression-free interval (C) than those with low ELFN1 expression. D-I: mRNA expression level of ELFN1 in COAD patients with different clinicopathological features. *P<0.05, **P<0.01, ***P<0.001.

表1 ELFN1在COAD中的单因素和多因素Cox回归分析

Tab.1 Univariate and multivariate Cox regression analyses of overall survival of COAD patients based on ELFN1 expression levels

Characteristics	Total (n)	Univariate analysis		Multivariate analysis
Characteristics	Total (n)	Hazard ratio (95%CI)	P	Hazard ratio (95%CI)	P
Age (year)	477
≤65	194	Reference
>65	283	1.610 (1.052-2.463)	0.028	3.417 (1.309-8.920)	0.012
T stage	476
T1&T2	94	Reference
T3&T4	382	3.072 (1.423-6.631)	0.004	0.661 (0.120-3.637)	0.634
N stage	477
N0	283	Reference
N1&N2	194	2.592 (1.743-3.855)	<0.001	0.174 (0.016-1.839)	0.146
M stage	414
M0	348	Reference
M1	66	4.193 (2.683-6.554)	<0.001	5.463 (1.807-16.518)	0.003
Pathologic stage	466
Stage I&Stage II	267	Reference
Stage III&Stage IV	199	2.947 (1.942-4.471)	<0.001	14.791(1.177-185.875)	0.037
Perineural invasion	181
No	135	Reference
Yes	46	1.940 (0.982-3.832)	0.056	0.736 (0.280-1.935)	0.534
Lymphatic invasion	433
No	265	Reference
Yes	168	2.450 (1.614-3.720)	<0.001	1.408 (0.557-3.556)	0.470
ELFN1	477
Low	238	Reference
High	239	1.683 (1.135-2.496)	0.010	3.284 (1.154-9.339)	0.026

表2 ELFN1在COAD中的Logistic回归分析

Tab.2 Logistic regression analysis of ELFN1

Characteristic	Total (n)	OR (95%CI)	P
Pathologic T stage (T3&T4 vs T1&T2)	477	1.831 (0.975-3.600)	0.069
Pathologic N stage (N1&N2 vs N0)	478	1.743 (1.078-2.851)	0.025
Pathologic M stage (M1 vs M0)	415	2.425 (1.286-4.572)	0.006
Pathologic stage (Stage III&IV vs Stage I&II)	467	1.787 (1.104-2.936)	0.020
Primary therapy outcome (PD&SD vs PR&CR)	250	2.834 (1.120-7.152)	0.025
Perineural invasion (Yes vs No)	181	2.675 (0.984-7.424)	0.044
Lymphatic invasion (Yes vs No)	434	2.823 (1.653-4.934)	<0.001

图4 ELFN1差异表达基因分析及PPI网络

Fig.4 Analysis of differentially expressed genes related to ELFN1 and construction of the PPI network. A: Volcano plot of differentially expressed genes (DEGs, |log2(FC)|>1, P.adj<0.05). B: The co-expression chord diagram showing the top 10 genes positively and negatively correlated with ELFN1. C: Molecular protein interaction network of ELFN1.

图5 ELFN1互作蛋白鉴定及基因功能注释分析

Fig.5 ELFN1 interaction protein identification and gene function annotation analysis. A: GO functional annotation analysis of biological process (BP), cellular component (CC) and molecular function (MF). B: Analysis of 10 KEGG signaling pathways related to ELFN1. C, D: The 10 GSEA pathways associated with ELFN1 in COAD.

图6 ELFN1在COAD细胞系中的表达及功能

Fig.6 Expression and function of ELFN1 in COAD cell lines. A, B: ELFN1 expression levels in NCM460, SW480, SW620, RKO, HCT116, LOVO, and CACO2 cells detected by qRT-PCR (A) and Western blotting (B). *P<0.05, **P<0.01, ***P<0.001 vs NCM460 cells. C: Expression levels of ELFN1 in 10 pairs of colon cancer tissue samples (*P<0.05, **P<0.01, ***P<0.001 vs NATs). D: Transwell assay of SW480 cells transfected with si-ELFN1. E: EDU proliferation assay of SW480 cells transfected with si-ELFN1. **P<0.01, ***P<0.001.

图7 ELFN1在结肠癌致癌信号通路分子机制研究

Fig.7 Molecular mechanism of ELFN1 in the oncogenic signaling pathway of colorectal cancer. A: Analysis of relevant carcinogenic pathways of ELFN1 in COAD. B: Western blotting for detecting protein expressions in COAD cells with ELFN1 knockdown. *P<0.05, ***P<0.001.

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