Pancreatic cancer cells overexpress gelsolin family-capping proteins, which contribute to their cell motility
- C C Thompson1,
- F J Ashcroft1,*,
- S Patel1,*,
- G Saraga1,
- D Vimalachandran1,
- W Prime2,
- F Campbell3,
- A Dodson3,
- R E Jenkins4,
- N R Lemoine5,
- T Crnogorac-Jurcevic5,
- H L Yin6,
- E Costello1
- 1Division of Surgery and Oncology, Royal Liverpool University Hospital, University of Liverpool, Liverpool, UK
- 2Cancer Tissue Bank Research Centre, Department of Pathology, University of Liverpool, Liverpool, UK
- 3Department of Pathology, University of Liverpool, Liverpool, UK
- 4Department of Pharmacology and Therapeutics, University of Liverpool, Liverpool, UK
- 5Cancer Research UK Molecular Oncology Unit, Barts and the London School of Medicine and Dentistry, London, UK
- 6University of Texas Southwestern Medical Centre, Dallas, Texas, USA
- Correspondence to:
Division of Surgery and Oncology, Royal Liverpool University Hospital, 5th Floor UCD Building, Daulby Street, University of Liverpool, Liverpool L68 3 GA, UK;
- Accepted 9 May 2006
- Revised 19 April 2006
- Published Online First 17 July 2006
Background: Previously, proteomic methods were applied to characterise differentially expressed proteins in microdissected pancreatic ductal adenocarcinoma cells.
Aims: To report that CapG and a related protein, gelsolin, which have established roles in cell motility, are overexpressed in metastatic pancreatic cancer; and to describe their pattern of expression in pancreatic cancer tissue and their effect on cell motility in pancreatic cancer cell lines.
Methods: CapG was identified by mass spectrometry and immunoblotting. CapG and gelsolin expression was assessed by immunohistochemical analysis on a pancreatic cancer tissue microarray and correlated with clinical and pathological parameters. CapG and gelsolin levels were reduced using RNA interface in Suit-2, Panc-1 and MiaPaCa-2 cells. Cell motility was assessed using modified Boyden chamber or wound-healing assays.
Results: Multiple isoforms of CapG were detected in pancreatic cancer tissue and cell lines. Immunohistochemical analysis of benign (n = 44 patients) and malignant (n = 69) pancreatic ductal cells showed significantly higher CapG staining intensity in nuclear (p<0.001) and cytoplasmic (p<0.001) compartments of malignant cells. Similarly, gelsolin immunostaining of benign (n = 24 patients) and malignant (n = 68 patients) pancreatic ductal cells showed higher expression in both compartments (both p<0.001). High nuclear CapG was associated with increased tumour size (p = 0.001). High nuclear gelsolin was associated with reduced survival (p = 0.01). Reduction of CapG or gelsolin expression in cell lines by RNAi was accompanied by significantly impaired motility.
Conclusions: Up regulation of these actin-capping proteins in pancreatic cancer and their ability to modulate cell motility in vitro suggest their potentially important role in pancreatic cancer cell motility and consequently dissemination.
- IQR, interquartile range
- MALDI-TOF, matrix-assisted laser desorption/ionisation-time-of–flight
- MTT, 3-(4, 5-dimethythiazol-2-yl-2, 5-diphenyltetrazolium bromide)
↵* These authors contributed equally to this work.
Published Online First 17 July 2006
Funding: This work is supported by grants from Cancer Research UK, North West Cancer Research Fund, The Medical Research Council and the National Institute of Health Burn Centre grant P05GM21681.
Competing interests: None.
Ethical approval: The study was conducted with ethical approval from Cheshire and Merseyside Health Authority (Hamilton House, 24 Pall Mall, Liverpool L3 6AL, UK), LREC ref 03/02/316A.