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image of Prolyl 4-Hydroxylase Beta Peptide Promotes Invasion, Migration, and Epithelial-Mesenchymal Transition through Activation of the Claudin-1/ AMPK/TGF-β1 Pathway in Bladder Cancer Cells

Abstract

Introduction

Prolyl 4-hydroxylase beta peptide (P4HB) is a novel diagnostic and prognostic marker associated with cancer progression and clinical outcomes, and it is upregulated in multiple types of cancer cells. However, the influence and potential mechanisms of P4HB on the migration, invasion, and epithelial-mesenchymal transition (EMT) of bladder cancer cells remain unclear. This study aims to clarify the role of P4HB in the migration, invasion, and EMT of bladder cancer cells and to explore its potential mechanism related to the Claudin-1/AMPK/TGF-β1 pathway.

Methods

The mRNA and protein expression levels of P4HB were examined in human ureteral epithelial cells (SV-HUC-1) and five bladder cancer cell lines (J82, T24, 5637, UM-UC-3, and RT4). Stable cell lines with P4HB overexpression and knockdown were constructed, and the effects of P4HB on migration, invasion, EMT, and the expression of EMT-related genes in bladder cancer cells were analyzed using wound healing assays, Transwell invasion assays, cellular morphology observations, real-time quantitative PCR, in-cell western blotting, western blotting, and enzyme-linked immunosorbent assays. Furthermore, Claudin-1 siRNA was transfected into P4HB-overexpressing cells to investigate its potential role in P4HB-induced invasion and EMT in bladder cancer cells.

Results

P4HB mRNA and protein expressions were significantly upregulated in human bladder cancer cell lines compared to those in ureteral epithelial cells. Cell migration, invasion, and EMT were significantly promoted in P4HB-overexpressing stable bladder cancer cells and suppressed in P4HB-knockdown cells. Furthermore, interference with P4HB downregulated EMT-related Claudin-1 mRNA and protein expressions and regulated the expression of downstream genes and proteins of Claudin-1. Moreover, interference of Claudin-1 with its siRNA significantly reversed the invasion and EMT induced by P4HB-overexpression, however, the effect of Claudin-1 siRNA was revised by TGF-β1 agonist and AMPK inhibitor.

Conclusion

P4HB promoted migration, invasion, and EMT of bladder cancer cells by activating the Claudin-1/AMPK/TGF-β1-related pathway.

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2026-01-22
2026-01-29

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Prolyl 4-Hydroxylase Beta Peptide Promotes Invasion, Migration, and Epithelial-Mesenchymal Transition through Activation of the Claudin-1/ AMPK/TGF-β1 Pathway in Bladder Cancer Cells
Bentham Science Publishers ; https://doi.org/10.2174/0109298665418763251124051527
/content/journals/ppl/10.2174/0109298665418763251124051527
/content/journals/ppl/10.2174/0109298665418763251124051527
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  • Article Type:     Research Article
Keywords: TGF-β1 ; EMT ; P4HB ; AMPK ; Claudin-1 ; bladder cancer cells

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