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Cellular and Molecular Biology

Hypoxia-inducible factor 3α1 increases epithelial-to-mesenchymal transition and iron uptake to drive colorectal cancer liver metastasis

British Journal of Cancer (2024)Cite this article

Subjects

Abstract

Background/objectives

Hypoxia-inducible factor (HIF)-3α1’s role in colorectal cancer (CRC) cells, especially its effects on epithelial-mesenchymal transition (EMT), zinc finger E-box binding homeobox 2 (ZEB2) gene expression, and iron metabolism, remains largely unstudied. This research sought to elucidate these relationships.

Methods

RNA-seq was conducted to investigate the impact of HIF-3α1 overexpression in CRC cells. Dual-luciferase reporter assays assessed the direct targeting of ZEB2 by HIF-3α1. Scratch assays measured changes in cell migration following HIF-3α1 overexpression and ZEB2 knockdown. The effects of HIF-3α1 overexpression on colon tumour growth and liver metastasis were examined in vivo. Iron chelation was used to explore the role of iron metabolism in HIF-3α1-mediated EMT and tumour growth.

Results

HIF-3α1 overexpression induced EMT and upregulated ZEB2 expression, enhancing cancer cell migration. ZEB2 knockdown reduced mesenchymal markers and cell migration. HIF-3α1 promoted colon tumour growth and liver metastasis, increased transferrin receptor (TFRC) expression and cellular iron levels, and downregulated HIF-1α, HIF-2α, and NDRG1. Iron chelation mitigated HIF-3α1-mediated EMT, tumour growth, and survival.

Conclusions

HIF-3α1 plays a critical role in colon cancer progression by promoting EMT, iron accumulation, and metastasis through ZEB2 and TFRC regulation, suggesting potential therapeutic targets in CRC.

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Fig. 1: Overexpression of HIF3A1 in colon cancer cells promotes expression of EMT signature genes.
Fig. 2: Validation of RNA-Seq results by qRT-PCR and western blot analysis.
Fig. 3: HIF3A is increased in CRC liver metastasis than primary tumour and is correlated with ZEB2 expression.
Fig. 4: HIF3A1 induces ZEB2 transcription in an HRE-dependent manner.
Fig. 5: HIF3A1 increases CRC liver metastasis in mice.
Fig. 6: HIF3A1 induces TFRC expression and increases iron content of CRC cells.
Fig. 7: Dp44mT reduces mesenchymal markers, cell migration and tumour growth in HIF3A1 overexpressing cells.

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Data availability

For original data, please contact xxue@salud.unm.edu.

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Funding

XX was supported by the National Institutes of Health (P20 GM130422, K01DK114390), a Research Scholar Grant from the American Cancer Society (RSG-18-050-01-NEC), Environmental Health and Toxicology Pilot Awards from UNM Center for Native Environmental Health Equity Research (P50 MD015706) and New Mexico Integrative Science Program Incorporating Research in Environmental Sciences (1P30ES032755), Shared Resources and Research Program Support Pilot Project Awards from UNM comprehensive cancer center (P30CA118100), a Cardiovascular and Metabolic Disease Research Program Pilot Project Grant from UNMHSC Office of Research Signature Programs and the Dedicated Health Research Funds from Research Allocation Committee at the University of New Mexico School of Medicine. LV was partially supported by a Predoctoral Fellowship from the NIAID-funded Biology of Infectious Disease and Inflammation program (T32AI007538). DMF was supported by Academic Science Education and Research Training (ASERT) from NIGMS (K12-GM088021).

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Authors and Affiliations

  1. Department of Biochemistry and Molecular Biology, University of New Mexico, Albuquerque, NM, USA

    Luke B. Villareal, Daniel M. Falcon & Xiang Xue

  2. Guangdong Provincial Key Laboratory of Microbial Culture Collection and Application, State Key Laboratory of Applied Microbiology Southern China, Institute of Microbiology, Guangdong Academy of Sciences, Guangzhou, China

    Liwei Xie

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XX designed the research, LBV and DMF performed the research, LX contributed vital analytical tools, and LBV and XX wrote the paper.

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Correspondence to Xiang Xue.

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All animal studies were carried out in accordance with the Institute of Laboratory Animal Resources guidelines and approved by the University Committee on the Use and Care of Animals at the University of New Mexico (Protocol# 20-201060-HSC).

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Villareal, L.B., Falcon, D.M., Xie, L. et al. Hypoxia-inducible factor 3α1 increases epithelial-to-mesenchymal transition and iron uptake to drive colorectal cancer liver metastasis. Br J Cancer (2024). https://doi.org/10.1038/s41416-024-02699-3

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