Zinc-Dependent Regulation of ZEB1 and YAP1 Coactivation Promotes Epithelial-Mesenchymal Transition Plasticity and Metastasis in Pancreatic Cancer.

Abstract

BACKGROUND:

Pancreatic cancer is characterized by extensive metastasis. Epithelial-mesenchymal transition (EMT) plasticity plays a critical role in tumor progression and metastasis by maintaining the transition between EMT and mesenchymal-epithelial transition states. Our aim is to understand the molecular events regulating metastasis and EMT plasticity in pancreatic cancer.

METHODS:

The interactions between a cancer-promoting zinc transporter ZIP4, a zinc-dependent EMT transcriptional factor ZEB1, a coactivator YAP1, and integrin α3 (ITGA3) were examined in human pancreatic cancer cells, clinical specimens, spontaneous mouse models (KPC and KPCZ) and orthotopic xenografts, and 3-dimensional spheroid and organoid models. Correlations between ZIP4, miR-373, and its downstream targets were assessed by RNA in situ hybridization and immunohistochemical staining. The transcriptional regulation of ZEB1, YAP1, and ITGA3 by ZIP4 was determined by chromatin immunoprecipitation, co-immunoprecipitation, and luciferase reporter assays.

RESULTS:

The Hippo pathway effector YAP1 is a potent transcriptional coactivator and forms a complex with ZEB1 to activate ITGA3 transcription through the YAP1/transcriptional enhanced associate domain (TEAD) binding sites in human pancreatic cancer cells and KPC-derived mouse cells. ZIP4 upregulated YAP1 expression via activation of miR-373 and inhibition of the YAP1 repressor large tumor suppressor 2 kinase (LATS2). Furthermore, upregulation of ZIP4 promoted EMT plasticity, cell adhesion, spheroid formation, and organogenesis both in human pancreatic cancer cells, 3-dimensional spheroid model, xenograft model, and spontaneous mouse models (KPC and KPCZ) through ZEB1/YAP1-ITGA3 signaling axis.

CONCLUSION:

We demonstrated that ZIP4 activates ZEB1 and YAP1 through distinct mechanisms. The ZIP4-miR-373-LATS2-ZEB1/YAP1-ITGA3 signaling axis has a significant impact on pancreatic cancer metastasis and EMT plasticity.

• Authors: Mingyang Liu, Yuqing Zhang, Jingxuan Yang, Hanxiang Zhan, Zhijun Zhou, Yuanyuan Jiang, Xiuhui Shi, Xiao Fan, Junxia Zhang, Wenyi Luo, Kar-Ming A Fung, Chao Xu, Michael S Bronze, Courtney W Houchen, Min Li
• Journal: Gastroenterology (Gastroenterology) Vol. 160 Issue 5 Pg. 1771-1783.e1 (04 2021) ISSN: 1528-0012 [Electronic] United States
• PMID: 33421513 (Publication Type: Journal Article, Research Support, N.I.H., Extramural, Research Support, Non-U.S. Gov't)
• Copyright: Copyright © 2021 AGA Institute. Published by Elsevier Inc. All rights reserved.

Chemical References

• Adaptor Proteins, Signal Transducing
• Cation Transport Proteins
• ITGA3 protein, human
• Integrin alpha3
• Itga3 protein, mouse
• MIRN373 microRNA, human
• MIRN373 microRNA, mouse
• MicroRNAs
• SLC39A4 protein, human
• Slc39a4 protein, mouse
• Transcription Factors
• YAP-Signaling Proteins
• YAP1 protein, human
• Yap1 protein, mouse
• ZEB1 protein, human
• ZEB1 protein, mouse
• Zinc Finger E-box-Binding Homeobox 1
• Zinc

Topics

• Adaptor Proteins, Signal Transducing (genetics, metabolism)
• Animals
• Cation Transport Proteins (genetics, metabolism)
• Cell Line, Tumor
• Cell Movement
• Cell Plasticity
• Epithelial-Mesenchymal Transition
• Gene Expression Regulation, Neoplastic
• Humans
• Integrin alpha3 (genetics, metabolism)
• Mice
• MicroRNAs (genetics, metabolism)
• Neoplasm Invasiveness
• Neoplasm Metastasis
• Pancreatic Neoplasms (genetics, metabolism, pathology)
• Signal Transduction
• Spheroids, Cellular
• Transcription Factors (genetics, metabolism)
• YAP-Signaling Proteins
• Zinc (metabolism)
• Zinc Finger E-box-Binding Homeobox 1 (genetics, metabolism)