CEP192过表达可作为胃癌患者不良预后的生物标志物并通过调控G2/M期关键蛋白的表达影响肿瘤细胞恶性增殖

doi:10.12122/j.issn.1673-4254.2024.11.10

南方医科大学学报 ›› 2024, Vol. 44 ›› Issue (11): 2137-2145.doi: 10.12122/j.issn.1673-4254.2024.11.10

• • 上一篇

CEP192过表达可作为胃癌患者不良预后的生物标志物并通过调控G2/M期关键蛋白的表达影响肿瘤细胞恶性增殖

张震¹^,²(), 鲁辉¹^,², 陈孝华¹^,², 王炼¹^,², 王子良¹^,², 王月月³^,⁴, 葛思堂¹^,⁴, 左芦根¹^,⁴()

^1.蚌埠医科大学第一附属医院，胃肠外科，安徽蚌埠 233004
^3.蚌埠医科大学第一附属医院，检验科，安徽蚌埠 233004
^4.蚌埠医科大学第一附属医院，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
^2.蚌埠医科大学研究生院，安徽蚌埠 233030

收稿日期:2024-07-11 出版日期:2024-11-20 发布日期:2024-11-29
通讯作者: 左芦根 E-mail:zhangzhencg0525@163.com;zuolugen@126.com
作者简介:张震，在读硕士研究生，E-mail: zhangzhencg0525@163.com
基金资助:
安徽省高校自然科学基金重点项目(KJ2020A0550);安徽省卫生健康科研项目(AHWJ2022a019);蚌埠医科大学2023年度研究生科研创新计划(Byycx23139)

CEP192 overexpression is correlated with poor prognosis of gastric cancer and promotes gastric cancer cell proliferation by regulating PLK1/CDK1/Cyclin B1 signaling

Zhen ZHANG¹^,²(), Hui LU¹^,², Xiaohua CHEN¹^,², Lian WANG¹^,², Ziliang WANG¹^,², Yueyue WANG³^,⁴, Sitang GE¹^,⁴, Lugen ZUO¹^,⁴()

^1.Department of Gastrointestinal Surgery, First Affiliated Hospital of Bengbu Medical University, Bengbu 233004, China
^3.Clinical Laboratory, First Affiliated Hospital of Bengbu Medical University, Bengbu 233004, China
^4.Anhui Provincial Key Laboratory of Basic and Translational Research of Inflammation-related Diseases, First Affiliated Hospital of Bengbu Medical University, Bengbu 233004, China
^2.Graduate School, Bengbu Medical University, Bengbu 233030, China

Received:2024-07-11 Online:2024-11-20 Published:2024-11-29
Contact: Lugen ZUO E-mail:zhangzhencg0525@163.com;zuolugen@126.com

摘要/Abstract

摘要：

目的探讨CEP192在胃癌中的表达情况及其预后价值，进一步研究其对胃癌细胞生物学功能的影响。方法利用公共数据库探究CEP192在胃癌组织中的表达程度，通过免疫组织化学技术进一步验证CEP192在胃癌中的具体表达状态。通过构建Kaplan-Meier （K-M）生存曲线，探究CEP192蛋白表达与胃癌患者长期预后的关联性。同时，利用单因素和多因素Cox回归分析模型，深入分析影响胃癌患者根治术后5年生存率的风险因素。通过受试者工作特征（ROC）曲线分析评估CEP192表达水平在预测胃癌患者术后5年生存率的价值。生物信息学功能富集预测CEP192在胃癌发生发展中可能参与的生物学过程。采用慢病毒干扰及过表达CEP192，CCK-8法检测CEP192对胃癌细胞增殖能力的影响，免疫印迹法检测CEP192对胃癌细胞G2/M期关键蛋白PLK1、CDK1、Cyclin B1的影响。将慢病毒稳定转染的各组细胞接种于裸鼠背部皮下进行裸鼠成瘤实验，在体观察CEP192对小鼠体质量、瘤体大小及G2/M期关键蛋白的影响。结果 CEP192在胃癌中高表达且与患者预后不良相关（P<0.05）。多因素Cox回归分析发现，CEP192高表达、CEA≥5 ng/mL、CA199≥37 IU/mL、T3-4期和N2-3期均为影响胃癌患者术后5年生存率的独立危险因素（P<0.05）。GO及REACTOME基因富集分析提示CEP192可能参与细胞周期等生物学过程，GSEA基因集富集分析结果显示CEP192对细胞周期过程呈现正向调控。CCK-8结果示干扰CEP192后MGC-803细胞的增殖能力减弱，过表达细胞增殖能力增强（P<0.05）。免疫印迹结果显示干扰CEP192后PLK1、CDK1、Cyclin B1蛋白表达水平下降，过表达呈上升趋势（P<0.05）。裸鼠皮下成瘤实验结果显示，与对照组相比，干扰CEP192小鼠体质量降低，肿瘤体积变小，PLK1、CDK1、Cyclin B1蛋白水平下调，过表达CEP192则呈现相反趋势（P<0.05）。结论 CEP192在胃癌组织中高表达，与患者的不良预后相关，其可能通过调控G2/M期关键蛋白的表达影响肿瘤细胞恶性增殖。

关键词: 胃癌, CEP192, 不良预后, 增殖, 细胞周期

Abstract:

Objective To investigate the correlation of CEP192 expression with prognosis of gastric cancer and biological behaviors of gastric cancer cells. Methods Public databases and clinical tissue samples were used to examine CEP192 expression level in gastric cancer. Kaplan-Meier survival curves, univariate and multivariate Cox regression analyses, ROC curves and bioinformatics analyses were used to explore the risk factors affecting the 5-year postoperative survival, the correlation of CEP192 expression level with the patients' survival, and its biological role in gastric cancer development. In gastric cancer MGC-803 cells with lentivirus-mediated CEP192 interference or overexpression, cell proliferation and expressions of PLK1, CDK1 and Cyclin B1 were examined with CCK-8 assay and Western blotting. The effects of CEP192 knockdown or overexpression on tumorigenesis of MGC-803 cells was observed in nude mice, and the expressions of PLK1, CDK1 and Cyclin B1 in the xenografts were detected. Results CEP192 was highly expressed in gastric cancer and associated with poor prognosis of the patients (P<0.05). High expression of CEP192, CEA ≥5 ng/mL, CA199 ≥37 IU/mL, T3-4 stage, and N2-3 stage were independent risk factors affecting the patients' 5-year postoperative survival (P<0.05). Bioinformatics analyses suggested that CEP192 was involved in several vital biological processes and positively regulated cell cycle progression. In MGC-803 cells, CEP192 knockdown significantly inhibited cell proliferation and lowered the expression levels of PLK1, CDK1, and Cyclin B1, while its overexpression produced the opposite effects. In the nude mouse models, CEP192 knockdown resulted in lowered tumorigenic potential of MGC-803 cells and decreased protein levels of PLK1, CDK1, and Cyclin B1 in the xenografts, while CEP192 overexpression in MGC-803 cells caused the opposite changes. Conclusion CEP192 overexpression is correlated with unfavorable outcomes of gastric cancer patients and promotes gastric cell proliferation by regulating the key proteins during G2/M phase transition.

Key words: gastric cancer, CEP192, poor prognosis, proliferation, cell cycle

张震, 鲁辉, 陈孝华, 王炼, 王子良, 王月月, 葛思堂, 左芦根. CEP192过表达可作为胃癌患者不良预后的生物标志物并通过调控G2/M期关键蛋白的表达影响肿瘤细胞恶性增殖[J]. 南方医科大学学报, 2024, 44(11): 2137-2145.

Zhen ZHANG, Hui LU, Xiaohua CHEN, Lian WANG, Ziliang WANG, Yueyue WANG, Sitang GE, Lugen ZUO. CEP192 overexpression is correlated with poor prognosis of gastric cancer and promotes gastric cancer cell proliferation by regulating PLK1/CDK1/Cyclin B1 signaling[J]. Journal of Southern Medical University, 2024, 44(11): 2137-2145.

图/表 10

图 1 CEP192在多种恶性肿瘤中高表达

Fig. 1 CEP192 is highly expressed in multiple cancers. A: Expression of CEP192 in common human cancers. B, C: Expression of CEP192 in gastric cancer. *P<0.05, **P<0.01, ***P<0.001 vs normal group.

图 2 CEP192在胃癌组织中高表达

Fig.2 Immunohistochemical detection of CEP192 expression and cellular localization in gastric cancer and adjacent tissues (A) and quantitative analysis of its expression levels (B). *P<0.05 vs adjacent tissues.

表1 胃癌组织中CEP192表达量与临床及病理参数间的关系

Tab.1 Relationship between CEP192 expression and clinicopathological parameters of gastric cancer patients

Factors	n	CEP192 expression		χ²	P
Factors	n	Low (n=54)	High (n=55)	χ²	P
Gender
Male	58	30 (51.7%)	28 (48.3%)	0.236	0.627
Female	51	24 (47.1%)	27 (52.9%)
Age (year)
<60	47	26 (55.3%)	21 (44.7%)	1.103	0.294
≥60	62	28 (45.2%)	34 (54.8%)
CEA (ng/mL)
<5	55	35 (63.6%)	20 (36.4%)	8.823	0.003
≥5	54	19 (35.2%)	35 (64.8%)
CA199 (IU/mL)
<37	63	41 (65.1%)	22 (34.9%)	14.418	<0.001
≥37	46	13 (28.3%)	33 (71.7%)
Cancer cell type
Adenocarcinoma	83	43 (51.8%)	40 (48.2%)	0.715	0.398
Others	26	11 (42.3%)	15 (57.7%)
Tumor size (cm)
<5	49	26 (53.1%)	23 (46.9%)	0.441	0.507
≥5	60	28 (46.7%)	32 (53.3%)
T stage
T1-T2	52	33 (63.5%)	19 (36.5%)	7.708	0.005
T3-T4	57	21 (36.8%)	36 (63.2%)
N stage
N0-N1	54	35 (64.8%)	19 (35.2%)	9.987	0.002
N2-N3	55	19 (34.5%)	36 (65.5%)

图3 CEP192表达对胃癌患者术后5年生存率的影响

Fig.3 Impact of CEP192 expression on 5-year survival rate of gastric cancer patients after surgery.

图 4 影响胃癌患者术后5年生存率的危险因素分析

Fig. 4 Analysis of risk factors affecting the 5-year postoperative survival rate of gastric cancer patients. A: Impact of preoperative serum CEA level on the 5-year survival rate of the patients. B: Impact of preoperative serum CA199 level on the patients' survival. C: Impact of tumor T stage on the patients' survival. D: Impact of tumor N stage on the patients' survival. E: Cox regression analysis of independent risk factors affecting the 5-year postoperative survival rate of gastric cancer patients.

图 5 CEP192表达量对胃癌患者根治术后5年生存率的预判价值

Fig.5 Predictive value of CEP192 expression for 5-year survival rate of gastric cancer patients after radical surgery.

图6 CEP192基因功能富集分析

Fig.6 Gene functional enrichment analysis of CEP192. A: GO enrichment analysis of CEP192 and its co-expressed genes. B: REACTOME enrichment analysis of CEP192 and co-expressed genes. C-H: CEP192 and GSEA enrichment of co-expressed genes.

图7 CEP192促进MGC-803细胞的增殖能力

Fig.7 CEP192 overexpression promotes proliferation of MGC-803 cells. A: Proliferation of MGC-803 cells with CEP192 knockdown. B: Proliferation of MGC-803 cells with CEP192 overexpression. *P<0.05 vs sh-NC group.

图8 CEP192促进细胞周期蛋白的表达

Fig.8 CEP192 overexpression promote expressions of cell cycle proteins. A: Western blotting for detecting expressions of PLK1, CDK1 and Cyclin B1 proteins. B, C: Expression levels PLK1, CDK1 and Cyclin B1 proteins. *P<0.05 vs sh-NC or LV-NC group.

图9 CEP192促进体内胃癌组织的增殖

Fig.9 CEP192 overexpression promotes proliferation of gastric cancer cells in nude mice. A: Tumor xenografts in different groups. B, C: Tumor weight of the nude mice. D, E: The tumor volume of the nude mice. F-H: Western blotting for detecting the expression levels of PLK1, CDK1 and Cyclin B1 proteins in the xenografts. *P<0.05 vs sh-NC or LV-NC group.

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CEP192过表达可作为胃癌患者不良预后的生物标志物并通过调控G2/M期关键蛋白的表达影响肿瘤细胞恶性增殖

CEP192 overexpression is correlated with poor prognosis of gastric cancer and promotes gastric cancer cell proliferation by regulating PLK1/CDK1/Cyclin B1 signaling

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