高表达SURF4通过抑制紧密连接蛋白表达促进胃癌细胞的恶性生物学行为

doi:10.12122/j.issn.1673-4254.2025.08.17

摘要/Abstract

摘要：

目的研究SURF4在胃癌中的表达情况及远期预后，进一步分析其影响胃癌细胞恶性生物学行为的可能作用途径。方法采用公共数据库初步验证SURF4在胃癌中表达高低及与不良预后之间的关系。回顾性分析蚌埠医科大学第一附属医院2016年1月~2019年10月共155例胃癌患者的信息，采用免疫组织化学方法检测癌和癌旁中SURF4表达水平，以SURF4表达量中位数（IOD=2.82）为界分为低表达组和高表达组，并分析与远期预后的关系。通过Cox比例风险模型筛选相关的独立预测因子，K-M生存曲线评估胃癌患者术后5年生存率。通过TISIBD数据库分析SURF4的表达水平与免疫细胞浸润方面的联系。采用GEO数据集筛选SURF4差异基因，利用GO和KEGG分析SURF4在胃癌中可能的分子机制及作用途径。在体外构建胃癌细胞系（HGC-27），并将SURF4分为上调组、下调组和2个对照组，采用CCK8、细胞划痕、Transwell、Western blotting实验证实对胃癌细胞增殖、迁移、侵袭和上皮间质转化（EMT）的影响。结果 GEPIA数据集及免疫组化结果提示，SURF4在胃癌中表达水平升高（P<0.05）。K-M生存分析显示SURF4过表达组患者术后5年生存期缩短（Log-rankχ²=38.749，P<0.001）。Cox回归分析表明胃癌的预后与肿瘤T3~4期、N2~3期及CEA≥5 μg/L、CA19-9≥37 kU/L密切相关（P<0.05）。TISIBD数据库分析SURF4的表达量与与活化B细胞、NK细胞及CD8+效应记忆T细胞呈现负相关（P<0.05）；与CD4⁺T细胞呈现正相关（P<0.05）。富集分析预测SUFR4可能通过紧密连接途径发生EMT参与胃癌恶性演变。Western blotting结果显示上调SURF4使E-cadherin表达降低，N-cadherin表达升高（P<0.05）。过表达SURF4可以抑制ZO-1和Claudin-1的发生（P<0.05）。CCK8、细胞划痕和Transwell结果显示敲高SURF4可促进HGC-27胃癌细胞的增殖、迁移和侵袭能力（P<0.05）。结论 SURF4在胃癌组织中呈现高表达并且与患者5年生存期缩短显著相关，可能通过抑制紧密连接蛋白进而参与上皮细胞间质化使肿瘤发生增殖、迁移和侵袭等恶性生物学行为。

关键词: 胃癌, SURF4, 预后, 紧密连接蛋白, 上皮间质转化

Abstract:

Objective To study the impact of SURF4 expression level on long-term prognosis of gastric cancer (GC) and biological behaviors of GC cells. Methods SURF4 expression level in GC and its association with long-term patient prognosis were analyzed using publicly available databases and in 155 GC patients with low and high SURF4 expressions detected immunohistochemically. The Cox proportional hazard model and Kaplan-Meier survival curves were used to analyze independent prognostic predictors of GC and the 5-year survival rate of the patients with different SURF4 expression levels. Informatics analyses were conducted to explore the correlation of SURF4 expression level with immune cell infiltration in GC, SURF4-related differential genes and their associated pathways. In cultured GC cell line HGC-27, the effects of SURF4 knockdown and overexpression on proliferation, migration, invasion and epithelial-mesenchymal transition (EMT) were investigated. Results Analysis of GEPIA dataset and immunohistochemical results suggested significant SURF4 overexpression in GC (P<0.05), which was associated with shortened 5-year survival time of the patients (χ²=38.749, P<0.001). The prognosis of GC was closely related to tumor stage T3-4, N2-3, CEA≥5 μg/L and CA19-9≥37 kU/L (P<0.05). SURF4 expression level was negatively correlated with activated B cells, NK cells and CD8⁺ effector memory T cells (P<0.05) and positively correlated with CD4⁺ T cells (P<0.05). GO and KEGG enrichment analysis suggested that SUFR4 may participate in GC carcinogenesis by promoting EMT through the tight junction pathway. In HGC-27 cells, SURF4 overexpression significantly decreased E-cadherin expression, increased N-cadherin expression, inhibited ZO-1 and claudin-1 expressions, and promoted cell proliferation, migration and invasion. Conclusion SURF4 is highly expressed in GC, and its overexpression is associated with a shortened 5-year survival of the patients possibly by enhancing tumor cell proliferation, migration and invasion via inhibiting tight junction proteins and promoting EMT.

Key words: gastric cancer, SURF4 high expression, prognosis, tight junction protein, epithelial-mesenchymal transition

王子良, 陈孝华, 杨晶晶, 严晨, 张志郅, 黄炳轶, 赵萌, 刘嵩, 葛思堂, 左芦根, 陈德利. 高表达SURF4通过抑制紧密连接蛋白表达促进胃癌细胞的恶性生物学行为[J]. 南方医科大学学报, 2025, 45(8): 1732-1742.

Ziliang WANG, Xiaohua CHEN, Jingjing YANG, Chen YAN, Zhizhi ZHANG, Bingyi HUANG, Meng ZHAO, Song LIU, Sitang GE, Lugen ZUO, Deli CHEN. High expression of SURF4 promotes migration, invasion and proliferation of gastric cancer cells by inhibiting tight junction proteins[J]. Journal of Southern Medical University, 2025, 45(8): 1732-1742.

图/表 11

图1 SURF4在肿瘤中的分布特征

Fig.1 Distribution characteristics of SURF4 in different cancers. A: Expression of SURF4 in different human cancers. B: Expression of SURF4 in gastric cancer. *P<0.05. C: Distribution characteristics of SURF4 in gastric cancer and adjacent tissues detected by immunohistochemistry. D: Relative IOD value of SURF4. n=155. *P<0.05 vs adjacent.

图2 UALCAN数据库分析不同病理参数下SURF4表达水平

Fig.2 Analysis of expression levels of SURF4 in gastric cancer patients with different pathological parameters based on UALCAN database. A: Sample type. B: Tumor grading. C: Cancer staging. D: Age. *P<0.05, **P<0.01, ***P<0.001 vs Normal.

图3 TISIBD数据平台分析SURF4的表达特征与免疫微环境的联系

Fig.3 Relationship between SURF4 expression level and immune microenvironment of gastric cancer analyzed using TISIBD data platform. A: Activated B cells. B: CD8+ effector memory T cells. C: NK cells. D: CD4+ thymocytes.

表1 胃癌SURF4表达水平与临床病理参数的联系

Tab.1 Relationship between SURF4 expression levels and clinicopathological parameters in 155 patients with gastric cancer ［n（%）］

Factors	n	SURF4 expression		χ²	P
Factors	n	Low (n=77)	High (n=78)	χ²	P
Gender				0.919	0.338
Male	114	54 (47.4)	60 (52.6)
Female	41	23 (56.1)	18 (43.9)
Age (year)				1.459	0.227
<60	65	36 (55.4)	29 (44.6)
≥60	90	41 (45.6)	49 (54.4)
CEA (μg/L)				10.874	<0.001
<5	80	50 (62.5)	30 (37.5)
≥5	75	27 (36.0)	48 (64.0)
CA19-9 (kU/L)				8.080	0.004
<37	87	52 (59.8)	35 (40.2)
≥37	68	25 (36.8)	43 (63.2)
Tumor size (cm)				0.156	0.693
<5	87	42 (48.3)	45 (51.7)
≥5	68	35 (51.5)	33 (48.5)
Histological type				1.455	0.228
Adenocarcinoma	112	59 (52.7)	53 (47.3)
Other	43	18 (41.9)	25 (58.1)
T stage				8.836	0.003
1-2	78	48 (61.5)	30 (38.5)
3-4	77	29 (37.7)	48 (62.3)
N stage				5.441	0.020
0-1	70	42 (60.0)	28 (40.0)
2-3	85	35 (41.2)	80 (58.8)

图4 SURF4表达量与胃癌根治术后5年生存期的关系

Fig.4 Relationship between SURF4 expression and 5-year survival of the patients after radical gastrectomy for gastric cancer. A: Relationship between SURF4 expression and overall survival. B: Postoperative survival of patients in high expression group and low expression group.

表2 Cox回归分析胃癌根治术后5年生存期的预后因子

Tab.2 Cox regression analysis of prognostic factors for 5-year survival after radical gastrectomy for gastric cancer

Factor	Univariate analysis		Multivariate analysis
Factor	Log rank χ²	P	HR	95% CI	P
Gender (Male vs female)	2.023	0.155	-	-	-
Age (≥60 years vs <60 years)	2.691	0.101	-	-	-
SURF4 expression (low vs high)	38.749	<0.001	3.106	1.853-5.206	<0.001
CEA (≥5 μg/L vs <5 μg/L)	34.667	<0.001	1.853	1.092-3.146	0.022
CA19-9 (≥37kU/L vs <37kU/L)	35.752	<0.001	1.949	1.125-3.378	0.017
Tumor size (≥5 cm vs <5 cm)	0.044	0.833	-	-	-
Histological type (Adenocarcinoma vs other)	0.013	0.910	-	-	-
T stage (T3-T4 vs T1-T2)	31.071	<0.001	2.774	1.661-4.635	<0.001
N stage (N2-N3 vs N0-N1)	28.779	<0.001	2.641	1.515-4.604	<0.001

图5 SURF4表达水平对胃癌患者术后5年生存情况的预估

Fig.5 Prediction of 5-year survival of patients with gastric cancer after operation based on SURF4 expression level.

图6 SURF4的富集分析结果

Fig.6 Enrichment analysis results of SURF4. A: KEGG analysis. B: GO analysis.

图7 SURF4对胃癌细胞增殖能力的影响

Fig.7 Effect of SURF4 expression level on proliferation of gastric cancer HGC-27 cells. A: Cell proliferation from day 1 to day 5. B: Comparison of cell proliferation on day 5 (n=4). LV: overexpression; Si: SiRNA. *P<0.05 vs Si-Control; #P<0.05 vs LV-Control.

图8 过表达SURF4与HGC-27胃癌细胞迁移和侵袭能力及EMT之间的关系

Fig.8 Effect of SURF4 knockdown and overexpression on migration, invasion and EMT of HGC-27 gastric cancer cells. A, D: Results of scratch assay. B, E: Effect of SURF4 knockdown and overexpression on migration ability of HGC-27 cells. C, F: Invasion ability of HGC-27 cells with SURF4 knockdown and overexpression. G-I: Expressions of E-cadherin and N-cadherin protein in HGC-27 cells with SURF4 knockdown and overexpression. *P<0.05 vs Si-Control; #P<0.05 vs LV-Control(n=4).

图9 高表达SURF4与紧密连接蛋白表达的关系

Fig.9 Effect of SURF4 knockdown and overexpression on tight junction protein expression in HGC-27 cells detected by Western blotting. A-C: Expressions of ZO-1 and Caudin-1 protein in HGC-27 gastric cancer cells. *P<0.05 vs Si-Control; #P<0.05 vs LV-Control (n=4).

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