A genetic variant conferred high expression of CAV2 promotes pancreatic cancer progression and associates with poor prognosis
A genetic variant conferred high expression of CAV2 promotes pancreatic cancer progression and associates with poor prognosis
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DOI：
10.1016/j.ejca.2021.04.008
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发表时间：
2021-05-08
影响因子：
8.4
通讯作者：
Miao, Xiaoping
中科院分区：
文献类型：
--
作者：
Zhu, Ying;Tian, Jianbo;Miao, Xiaoping
目的：本研究旨在找出与胰腺导管腺癌预后相关的功能基因和基因变异，并揭示其影响预后的机制。方法：首先，我们对1070例患者进行两阶段外显子相关研究，以确定与胰腺导管腺癌预后相关的基因变异。然后，我们通过生物信息学分析和双荧光素酶报告基因分析进行精细定位，以揭示原因、功能变异和预后基因。接下来，我们分别用小干扰RNA、CRISPR/Cas9和慢病毒建立了基因敲除、基因敲除和基因过表达细胞系，并研究了基因在体内和体外对细胞增殖和迁移的作用。结果：我们发现rs8940标记的CAV1-CAV2基因座与PDAC预后显著相关，而CAV2基因3‘非翻译区的rs10249656才是真正的功能变异体，它通过取消miR-548s结合上调了CAV2的表达。我们观察到在PDAC中CAV2表达上调，高表达与预后不良相关。CAV2的瞬时敲除抑制了PDAC的迁移，但不影响细胞的增殖率。CAV2基因敲除抑制了PDAC的进展和转移，而稳定的CAV2过表达促进了PDAC的进展和转移。结论：CAV2在PDAC的进展中起重要作用，CAV2基因的遗传变异对PDAC的预后有重要影响。(C)2021年爱思唯尔有限公司。保留所有权利。
Aim: This study aimed to identify the functional genes and genetic variants associated with the prognosis of pancreatic ductal adenocarcinoma (PDAC) and reveal the mechanism underlying their prognostic roles.Methods: First, we implement a two-stage exome-wide association study in a total of 1070 patients to identify the genetic variant correlated with PDAC prognosis. Then we performed fine mapping through bioinformatics analysis and dual-luciferase reporter assays to reveal the causal functional variant and prognostic gene. Next, we established the gene knockdown, knockout, and overexpression cell lines with small interfering RNA, CRISPR/Cas9, and lentivirus, respectively, and investigated the gene function on cell proliferation and migration in vivo and in vitro. Finally, we performed the RNA-seq to elucidate downstream genes and mechanisms altering PDAC prognosis.Results: We identified the CAV1-CAV2 locus tagged by rs8940 was significantly associated with PDAC prognosis, and rs10249656 in the 3'untranslated region of CAV2 was the real functional variant, which upregulated CAV2 expression through abolishing miR-548s binding. We observed upregulated CAV2 in PDAC and the higher expression correlated with worse prognosis. Transient knockdown of CAV2 inhibited PDAC migration without affecting proliferation rate. Knockout of CAV2 suppressed PDAC progression and metastasis, whereas stable overexpression of CAV2 promoted. Overexpressed CAV2 promoted the PDAC progression and metastasis via perturbing genes in the focal adhesion (CCND1, IGTA1, and ZYX) and extracellular matrix organisation (PLOD2, CAST, and ITGA1) pathways mechanically.Conclusion: These findings shed light on an important role of CAV2 on PDAC progression and the prognostic impact of its genetic variation. (C) 2021 Elsevier Ltd. All rights reserved.