TCF7L2 and EGR1 synergistic activation of transcription of LCN2 via an ERK1/2-dependent pathway in esophageal squamous cell carcinoma cells

Yan Zhao; Qiaoxi Xia; Yan Liu; Wenjing Bai; Yubin Yao; Jiyu Ding; Ling Lin; Zhennan Xu; Zhixiong Cai; Shaohong Wang; Enmin Li; Haixiong Xu; Bingli Wu; Liyan Xu; Zepeng Du

doi:10.1016/j.cellsig.2018.12.007

TCF7L2 and EGR1 synergistic activation of transcription of LCN2 via an ERK1/2-dependent pathway in esophageal squamous cell carcinoma cells

Cell Signal. 2019 Mar:55:8-16. doi: 10.1016/j.cellsig.2018.12.007. Epub 2018 Dec 14.

Authors

Yan Zhao¹, Qiaoxi Xia², Yan Liu³, Wenjing Bai², Yubin Yao⁴, Jiyu Ding², Ling Lin², Zhennan Xu³, Zhixiong Cai⁵, Shaohong Wang⁶, Enmin Li², Haixiong Xu³, Bingli Wu⁷, Liyan Xu⁸, Zepeng Du⁹

Affiliations

¹ Department of Pathology, Shantou Central Hospital, Affiliated Shantou Hospital of Sun Yat-sen University, Shantou 515041, China; Key Laboratory of Molecular Biology in High Cancer Incidence Coastal Chaoshan Area of Guangdong Higher Education Institutes, Shantou University Medical College, Shantou 515041, China; Institute of Oncologic Pathology, Shantou University Medical College, Shantou 515041, China.
² Key Laboratory of Molecular Biology in High Cancer Incidence Coastal Chaoshan Area of Guangdong Higher Education Institutes, Shantou University Medical College, Shantou 515041, China; Department of Biochemistry and Molecular Biology, Shantou University Medical College, Shantou 515041, China.
³ Department of Neurosurgery, Shantou Central Hospital, Affiliated Shantou Hospital of Sun Yat-sen University, Shantou 515041, China.
⁴ Department of Radiology, Shantou Central Hospital, Affiliated Shantou Hospital of Sun Yat-sen University, Shantou 515041, China.
⁵ Department of Cardiology, Shantou Central Hospital, Affiliated Shantou Hospital of Sun Yat-sen University, Shantou 515041, China.
⁶ Department of Pathology, Shantou Central Hospital, Affiliated Shantou Hospital of Sun Yat-sen University, Shantou 515041, China.
⁷ Key Laboratory of Molecular Biology in High Cancer Incidence Coastal Chaoshan Area of Guangdong Higher Education Institutes, Shantou University Medical College, Shantou 515041, China; Department of Biochemistry and Molecular Biology, Shantou University Medical College, Shantou 515041, China. Electronic address: blwu@stu.edu.cn.
⁸ Key Laboratory of Molecular Biology in High Cancer Incidence Coastal Chaoshan Area of Guangdong Higher Education Institutes, Shantou University Medical College, Shantou 515041, China; Institute of Oncologic Pathology, Shantou University Medical College, Shantou 515041, China. Electronic address: lyxu@stu.edu.cn.
⁹ Department of Pathology, Shantou Central Hospital, Affiliated Shantou Hospital of Sun Yat-sen University, Shantou 515041, China; Guangdong Province Key Laboratory of Malignant Tumor Epigenetics and Genes Regulation, Sun Yat-sen Memorial Hospital, Sun Yat-sen University, Guangzhou 510120, China. Electronic address: zepdu@126.com.

PMID: 30557604
DOI: 10.1016/j.cellsig.2018.12.007

Abstract

High level expression of lipocalin 2 (LCN2) usually indicates poor prognosis in esophageal squamous cell carcinoma (ESCC) and many other cancers. Our previous study showed LCN2 promotes migration and invasion of ESCC cells through a novel positive feedback loop. However, the key transcription activation protein (KTAP) in the loop had not yet been identified. In this study, we first predicted the most probable KTAPs by bioinformatic analysis. We then assessed the transcription regulatory regions in the human LCN2 gene by fusing deletions of its 5'-flanking region to a dual-luciferase reporter. We found that the region -720/-200 containing transcription factor 7-like 2 (TCF7L2) (-273/-209) and early growth response 1 (EGR1) (-710/-616) binding sites is crucial for LCN2 promoter activity. Chromatin immunoprecipitation (ChIP) experiments demonstrated that TCF7L2 and EGR1 bound directly to their binding sites within the LCN2 promoter as KTAPs. Mechanistically, overexpression of TCF7L2 and EGR1 increased endogenous LCN2 expression via the ERK signaling pathway. Treatment with recombinant human LCN2 protein enhanced activation of the ERK pathway to facilitate endogenous LCN2 expression, as well as increase the expression level of TCF7L2 and EGR1. Treatment with the MEK inhibitor U0126 inhibited the activation by TCF7L2 or EGR1 overexpression. Moreover, overexpression of TCF7L2 or EGR1 accelerated the migration and invasion of ESCC cells. A synergistic effect was observed between TCF7L2 and EGR1 in amplifying the induction of LCN2 and enhancing migration and invasion. Taken together, our study indicates that TCF7L2 and EGR1 are the KTAPs of LCN2, within a positive "LCN2 → MEK/ERK → LCN2" path, to promote the migration and invasion of ESCC cells. Based on their clinicopathological significance, LCN2 and its two expression regulators TCF7L2 and ERG1 might be therapeutic targets for ESCC.

Keywords: EGR1; Esophageal squamous cell carcinoma; Key transcription activation protein; LCN2; TCF7L2.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Cell Line, Tumor
Cell Movement
Early Growth Response Protein 1 / physiology*
Esophageal Neoplasms* / genetics
Esophageal Neoplasms* / metabolism
Esophageal Squamous Cell Carcinoma* / genetics
Esophageal Squamous Cell Carcinoma* / metabolism
Gene Expression Regulation, Neoplastic
Humans
Lipocalin-2 / metabolism*
MAP Kinase Signaling System
Promoter Regions, Genetic
Transcription Factor 7-Like 2 Protein / physiology*

Substances

EGR1 protein, human
Early Growth Response Protein 1
LCN2 protein, human
Lipocalin-2
TCF7L2 protein, human
Transcription Factor 7-Like 2 Protein