蜗牛/叉头框蛋白K1/富含半胱氨酸的血管生成素61信号轴调控结直肠癌的上皮-间质转化和转移

Snail/FOXK1/Cyr61 Signaling Axis Regulates the Epithelial-Mesenchymal Transition and Metastasis in Colorectal Cancer.

作者信息

Huang Xiaoting, Xiang Li, Li Yueqiao, Zhao Yingying, Zhu Huiqiong, Xiao Yizhi, Liu Mengwei, Wu Xiaosheng, Wang Zhiqing, Jiang Ping, Qing Haitao, Zhang Qiang, Liu Guangnan, Zhang Wenjing, Li Aimin, Chen Ye, Liu Side, Wang Jide

机构信息

Guangdong Provincial Key Laboratory of Gastroenterology, Department of Gastroenterology, Nanfang Hospital, Southern Medical University, Guangzhou, China.

Department of Gastroenterology, Longgang District People's Hospital, Shenzhen, China.

出版信息

Cell Physiol Biochem. 2018;47(2):590-603. doi: 10.1159/000490015. Epub 2018 May 22.

DOI:10.1159/000490015

PMID:29794466

Abstract

BACKGROUND/AIMS: Metastasis is the primary cause of colorectal cancer (CRC)-related death. However, the molecular mechanisms underlying metastasis in CRC remain unclear.

METHODS

We evaluated mRNA and protein expression levels by quantitative real-time reverse transcription PCR, western blotting, immunofluorescence, tissue microarrays, and immunohistochemistry assays. We also assessed the migration and invasion abilities of CRC cells in vitro by wound healing assays, invasion and migration assays, western blot analysis, and immunofluorescence. Tumor metastasis was evaluated in nude mice in vivo.

RESULTS

A positive correlation was observed between the expression patterns of Forkhead box k1 (FOXK1) and Snail in CRC. Luciferase reporter and chromatin immunoprecipitation assays demonstrated that Snail directly bound to and activated the human FOXK1 gene promoter. Moreover, the Snail-FOXK1 axis promote epithelial mesenchymal transition (EMT)-mediated CRC cell invasion and metastasis. FOXK1 and Snail expression levels were correlated with tumor progression and served as significant predictors of overall survival in patients with CRC. Furthermore, overexpression of FOXK1 induced the EMT by upregulating the expression of cysteine-rich angiogenic inducer 61 (Cyr61). Luciferase assays showed that Cyr61 was a direct transcriptional target of FOXK1. Down regulation of Cyr61 decreased FOXK1-enhanced "CRC cell" migration, invasion, and metastasis. Additionally, FOXK1 expression was positively correlated with Cyr61 expression and was associated with poor prognosis.

CONCLUSIONS

The Snail/FOXK1/Cyr61 signaling axis regulates the EMT and metastasis of CRC.

摘要

背景/目的：转移是结直肠癌（CRC）相关死亡的主要原因。然而，CRC转移的分子机制仍不清楚。

方法

我们通过定量实时逆转录PCR、蛋白质印迹、免疫荧光、组织芯片和免疫组织化学分析评估mRNA和蛋白质表达水平。我们还通过伤口愈合试验、侵袭和迁移试验、蛋白质印迹分析和免疫荧光在体外评估CRC细胞的迁移和侵袭能力。在裸鼠体内评估肿瘤转移情况。

结果

在CRC中观察到叉头框k1（FOXK1）和Snail的表达模式之间呈正相关。荧光素酶报告基因和染色质免疫沉淀试验表明，Snail直接结合并激活人FOXK1基因启动子。此外，Snail-FOXK1轴促进上皮-间质转化（EMT）介导的CRC细胞侵袭和转移。FOXK1和Snail表达水平与肿瘤进展相关，并可作为CRC患者总生存期的重要预测指标。此外，FOXK1的过表达通过上调富含半胱氨酸的血管生成诱导因子61（Cyr61）的表达诱导EMT。荧光素酶试验表明，Cyr61是FOXK1的直接转录靶点。下调Cyr61可降低FOXK1增强的“CRC细胞”迁移、侵袭和转移。此外，FOXK1表达与Cyr61表达呈正相关，且与预后不良有关。