Projects per year
Abstract
Objectives
Cell motility is fundamental to thyroid cancer progression, integrating complex signalling networks, cell adhesion, and actin dynamics. The proto-oncogene pituitary tumor-transforming gene (PTTG)-binding factor (PBF/PTTG1IP) is upregulated in thyroid cancer and associated with tumour progression. PBF promotes thyroid cancer cell migration and invasion through Y174 phosphorylation by Src kinase and phosphoproteomic and RNA-Seq analyses of Nthy-ori 3-1 thyroid cells revealed PBF overexpression altered regulation of adhesion and cytoskeletal proteins. We hypothesised that oncogenic PBF expression promotes thyroid cancer cell motility through altering adhesion dynamics.
Methods: We utilised CRISPR/Cas9-mediated PBF-knockout (KO) TPC-1 human papillary thyroid carcinoma cells to quantitatively assess cell adhesion and spreading kinetics on fibronectin
Results: Cell adhesion assays demonstrated that PBF-KO TPC-1 cells exhibited significantly decreased adhesion and spreading on fibronectin compared with control TPC-1 cells. Immunofluorescence staining of focal adhesions (FAs), which connect the cytoskeleton and extracellular matrix, revealed markedly fewer, shorter FAs predominantly around the cell periphery in PBF-depleted TPC-1 cells, compared with control cells that displayed numerous, elongated FAs along actin fibres. The expression of FA proteins focal adhesion kinase (FAK), vinculin and paxillin remained unchanged in PBF KO cells whereas FAK-Y397 phosphorylation was significantly decreased. Early adhesion assays (15-60 min) subsequently demonstrated delayed FAK-Y397 phosphorylation and FA development in PBF-KO TPC-1 cells, suggesting diminished FAK activation. Live cell LifeAct-GFP (F-actin) imaging indicated impaired lamellipodia formation and lack of directionality in PBF-KO TPC-1 cells. During migration, PBF-KO TPC-1 cells displayed disorganised Golgi, lacking orientation towards the leading edge, suggesting impaired polarity.
Conclusions
These findings identify a critical role for PBF in promoting thyroid cancer cell motility by regulating early focal adhesion assembly and maturation, as well as maintaining cellular polarity necessary for directed migration. Elucidating the precise molecular mechanisms by which PBF interacts with the adhesion machinery may unveil novel therapeutic avenues for targeting thyroid cancer progression.
Cell motility is fundamental to thyroid cancer progression, integrating complex signalling networks, cell adhesion, and actin dynamics. The proto-oncogene pituitary tumor-transforming gene (PTTG)-binding factor (PBF/PTTG1IP) is upregulated in thyroid cancer and associated with tumour progression. PBF promotes thyroid cancer cell migration and invasion through Y174 phosphorylation by Src kinase and phosphoproteomic and RNA-Seq analyses of Nthy-ori 3-1 thyroid cells revealed PBF overexpression altered regulation of adhesion and cytoskeletal proteins. We hypothesised that oncogenic PBF expression promotes thyroid cancer cell motility through altering adhesion dynamics.
Methods: We utilised CRISPR/Cas9-mediated PBF-knockout (KO) TPC-1 human papillary thyroid carcinoma cells to quantitatively assess cell adhesion and spreading kinetics on fibronectin
Results: Cell adhesion assays demonstrated that PBF-KO TPC-1 cells exhibited significantly decreased adhesion and spreading on fibronectin compared with control TPC-1 cells. Immunofluorescence staining of focal adhesions (FAs), which connect the cytoskeleton and extracellular matrix, revealed markedly fewer, shorter FAs predominantly around the cell periphery in PBF-depleted TPC-1 cells, compared with control cells that displayed numerous, elongated FAs along actin fibres. The expression of FA proteins focal adhesion kinase (FAK), vinculin and paxillin remained unchanged in PBF KO cells whereas FAK-Y397 phosphorylation was significantly decreased. Early adhesion assays (15-60 min) subsequently demonstrated delayed FAK-Y397 phosphorylation and FA development in PBF-KO TPC-1 cells, suggesting diminished FAK activation. Live cell LifeAct-GFP (F-actin) imaging indicated impaired lamellipodia formation and lack of directionality in PBF-KO TPC-1 cells. During migration, PBF-KO TPC-1 cells displayed disorganised Golgi, lacking orientation towards the leading edge, suggesting impaired polarity.
Conclusions
These findings identify a critical role for PBF in promoting thyroid cancer cell motility by regulating early focal adhesion assembly and maturation, as well as maintaining cellular polarity necessary for directed migration. Elucidating the precise molecular mechanisms by which PBF interacts with the adhesion machinery may unveil novel therapeutic avenues for targeting thyroid cancer progression.
| Original language | English |
|---|---|
| Article number | Poster 174 |
| Pages (from-to) | e39-e39 |
| Number of pages | 1 |
| Journal | Thyroid |
| Volume | 35 |
| Issue number | S1 |
| DOIs | |
| Publication status | Published - 12 Sept 2025 |
| Event | American Thyroid Association 2025 Annual Meeting - The Westin Kierland, Scottsdale, United States Duration: 10 Sept 2025 → 14 Sept 2025 https://www.thyroid.org/2025-annual-meeting/ |
Access to Document
Fingerprint
Dive into the research topics of 'PBF-Mediated Focal Adhesion Dynamics: A Key Driver of Thyroid Cancer Cell Motility'. Together they form a unique fingerprint.-
TRANSFORMING CELLULAR IMAGING AND ABLATION STRATEGIES VIA NEW MECHANISTIC UNDERSTANDING OF THE SODIUM IODIDE SYMPORTER
McCabe, C. (Principal Investigator)
1/12/24 → 31/05/28
Project: Research Councils
-
PBF Signalling in the Regulation of Cell Motility
Smith, V. (Principal Investigator)
Biotechnology & Biological Sciences Research Council
1/06/21 → 12/09/25
Project: Research Councils