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The Roles of ZnT1 and ZnT4 in Glucose-Stimulated Zinc Secretion in Prostate Epithelial Cells

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Abstract

Purpose

We have previously demonstrated by MRI that high glucose stimulates efflux of zinc ions from the prostate. To our knowledge, this phenomena had not been reported previously and the mechanism remains unknown. Here, we report some initial observations that provide new insights into zinc processing during glucose-stimulated zinc secretion (GSZS) in the immortalized human prostate epithelial cell line, PNT1A. Additionally, we identified the subtypes of zinc-containing cells in human benign prostatic hyperplasia (BPH) tissue to further identify which cell types are likely responsible for zinc release in vivo.

Procedure

An intracellular fluorescence marker, FluoZin-1-AM, was used to assess the different roles of ZnT1 and ZnT4 in zinc homeostasis in wild type (WT) and mRNA knockdown PNT1A cell lines. Additionally, Bafilomycin A1 (Baf) was used to disrupt lysosomes and assess the role of lysosomal storage during GSZS. ZIMIR, an extracellular zinc-responsive fluorescent marker, was used to assess dynamic zinc efflux of WT and ZnT1 mRNA knockdown cells exposed to high glucose. Electron microscopy was used to assess intracellular zinc storage in response to high glucose and evaluate how Bafilomycin A1 affects zinc trafficking. BPH cells were harvested from transurtheral prostatectomy tissue and stained with fluorescent zinc granule indicator (ZIGIR), an intracellular zinc-responsive fluorescent marker, before being sorted for cell types using flow cytometry.

Results

Fluorescent studies demonstrate that ZnT1 is the major zinc efflux transporter in prostate epithelial cells and that loss of ZnT1 via mRNA knockdown combined with lysosomal storage disruption results in a nearly 4-fold increase in cytosolic zinc. Knockdown of ZnT1 dramatically reduces zinc efflux during GSZS. Electron microscopy (EM) reveals that glucose stimulation significantly increases lysosomal storage of zinc; disruption of lysosomes via Baf or ZnT4 mRNA knockdown increases multi-vesicular body (MVB) formation and cytosolic zinc levels. In human BPH tissue, only the luminal epithelial cells contained significant amounts of zinc storage granules.

Conclusions

Exposure of prostate epithelial cells to high glucose alters zinc homeostasis by inducing efflux of zinc ions via ZnT1 channels and increasing lysosomal storage via ZnT4. Given that prostate cancer cells undergo profound metabolic changes that result in reduced levels of total zinc, understanding the complex interplay between glucose exposure and zinc homeostasis in the prostate may provide new insights into the development of prostate carcinogenesis.

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Acknowledgements

We thank Dr. Wen-hong Li (Department of Cell Biology, UT Southwestern Medical Center) for the gift of the ZIMIR and critical reading of the manuscript.

Funding

This work was supported by grants from the National Institutes of Health (R01-DK095416), the Robert A. Welch Foundation (AT-584), and the Cancer Prevention & Research Institute of Texas (RP-180178).

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Authors and Affiliations

  1. Advanced Imaging Research Center, The University of Texas Southwestern Medical Center, 5323 Harry Hines Blvd, NE 4.210, Dallas, TX, 75390-8568, USA

    Su-Tang Lo, Daniel Parrott, M. Veronica Clavijo Jordan & A. Dean Sherry

  2. Department of Radiology, UT Southwestern Medical Center, Dallas, TX, 75390-8896, USA

    Su-Tang Lo, Daniel Parrott, M. Veronica Clavijo Jordan & A. Dean Sherry

  3. Athinoula A. Martinos Center for Biomedical Imaging, Massachusetts General Hospital, Harvard Medical School, Charlestown, MA, 02129, USA

    M. Veronica Clavijo Jordan, Diya Binoy Joseph & Douglas Strand

  4. Department of Urology, UT Southwestern Medical Center, Dallas, TX, 75390-9110, USA

    U-Ging Lo

  5. Department of Life Sciences, National Chung Hsing University, Taichung City, 402, Taiwan

    Ho Lin

  6. Electron Microscopy Core Facility, UT Southwestern Medical Center, Dallas, TX, 75390-9039, USA

    Anza Darehshouri

  7. Department of Chemistry, University of Texas at Dallas, Richardson, TX, 75083, USA

    A. Dean Sherry

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  1. Su-Tang Lo
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Correspondence to A. Dean Sherry.

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Conflict of Interest

A.D.S. is a co-founder of VitalQuan, LLC, which is developing MRI and fluorescence sensors for detecting Zn2+ in tissues.

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Lo, ST., Parrott, D., Jordan, M.V.C. et al. The Roles of ZnT1 and ZnT4 in Glucose-Stimulated Zinc Secretion in Prostate Epithelial Cells. Mol Imaging Biol 23, 230–240 (2021). https://doi.org/10.1007/s11307-020-01557-x

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