神经黏附分子 4 的表达与嗜铬细胞瘤和副神经节瘤的恶性行为有关。

Expression of Contactin 4 Is Associated With Malignant Behavior in Pheochromocytomas and Paragangliomas.

机构信息

Pole of Cardiovascular Research, Institut de Recherche Expérimentale et Clinique, Université catholique de Louvain, Brussels, Belgium.

Department of Pathology, Erasmus MC Cancer Institute, University Medical Center, Rotterdam, Netherlands.

出版信息

J Clin Endocrinol Metab. 2018 Jan 1;103(1):46-55. doi: 10.1210/jc.2017-01314.

DOI:10.1210/jc.2017-01314

PMID:28938490

Abstract

CONTEXT

Pheochromocytomas and paragangliomas (PPGLs) are rare neuroendocrine, usually benign, tumors. Currently, the only reliable criterion of malignancy is the presence of metastases.

OBJECTIVE

The aim of this study was to identify genes associated with malignancy in PPGLs.

DESIGN

Transcriptomic profiling was performed on 40 benign and 11 malignant PPGLs. Genes showing a significantly different expression between benign and malignant PPGLs with a ratio ≥4 were confirmed and tested in an independent series by quantitative real-time polymerase chain reaction (qRT-PCR). Immunohistochemistry was performed for the validated genes on 109 benign and 32 malignant PPGLs. Functional assays were performed with hPheo1 cells.

SETTING

This study was conducted at the Department of Pathology of the Erasmus MC University Medical Center Rotterdam Human Molecular Genetics laboratory of the de Duve Institute, University of Louvain.

PATIENTS

PPGL samples from 179 patients, diagnosed between 1972 and 2015, were included.

MAIN OUTCOME MEASURES

Associations between gene expression and malignancy were tested using supervised clustering approaches.

RESULTS

Ten differentially expressed genes were selected based on messenger RNA (mRNA) expression array data. Contactin 4 (CNTN4) was overexpressed in malignant vs benign tumors [4.62-fold; false discovery rate (FDR), 0.001]. Overexpression at the mRNA level was confirmed using qRT-PCR (2.90-fold, P = 0.02; validation set: 4.26-fold, P = 0.005). Consistent findings were obtained in The Cancer Genome Atlas cohort (2.7-fold; FDR, 0.02). CNTN4 protein was more frequently expressed in malignant than in benign PPGLs by immunohistochemistry (58% vs 17%; P = 0.002). Survival after 7 days of culture under starvation conditions was significantly enhanced in hPheo1 cells transfected with CNTN4 complementary DNA.

CONCLUSION

CNTN4 expression is consistently associated with malignant behavior in PPGLs.

摘要

背景

嗜铬细胞瘤和副神经节瘤（PPGLs）是罕见的神经内分泌肿瘤，通常为良性。目前，唯一可靠的恶性肿瘤标准是存在转移。

目的

本研究旨在鉴定与 PPGL 恶性相关的基因。

设计

对 40 例良性和 11 例恶性 PPGL 进行转录组谱分析。对良性和恶性 PPGL 之间表达差异显著（倍数≥4）的基因进行确认，并通过实时定量聚合酶链反应（qRT-PCR）在独立的系列中进行测试。对 109 例良性和 32 例恶性 PPGL 进行了验证基因的免疫组织化学检测。对 hPheo1 细胞进行了功能检测。

地点

该研究在鹿特丹伊拉斯姆斯大学医学中心的 Erasmus MC 大学病理学系、鲁汶大学的 de Duve 研究所的人类分子遗传学实验室进行。

患者

纳入了 1972 年至 2015 年间诊断的 179 例患者的 PPGL 样本。

主要观察指标

采用监督聚类方法检测基因表达与恶性肿瘤之间的相关性。

结果

基于信使 RNA（mRNA）表达谱数据，选择了 10 个差异表达基因。与良性肿瘤相比，恶性肿瘤中接触蛋白 4（CNTN4）表达上调[4.62 倍；假发现率（FDR），0.001]。使用 qRT-PCR 验证了 mRNA 水平的过表达（2.90 倍，P=0.02；验证集：4.26 倍，P=0.005）。在癌症基因组图谱队列中也得到了一致的结果（2.7 倍；FDR，0.02）。CNTN4 蛋白在恶性 PPGL 中的表达频率明显高于良性 PPGL（58%对 17%；P=0.002）。hPheo1 细胞转染 CNTN4 cDNA 后，在饥饿条件下培养 7 天后的生存能力明显增强。