Phospholipase-C gamma-1 (PLCγ-1) is critical in hepatocyte growth factor induced in vitro invasion and migration without affecting the growth of prostate cancer cells

Davies, Gaynor; Martin, Tracey A.; Ye, Lin; Lewis-Russell, Jonathon M.; Mason, Malcolm D.; Jiang, Wen G.

doi:10.1016/j.urolonc.2007.06.003

« Previous Next »Urologic Oncology: Seminars and Original Investigations
Volume 26, Issue 4 , Pages 386-391, July 2008

Phospholipase-C gamma-1 (PLCγ-1) is critical in hepatocyte growth factor induced in vitro invasion and migration without affecting the growth of prostate cancer cells

Gaynor Davies, Ph.D.
- Metastasis and Angiogenesis Research Group, Department of Surgery, School of Medicine, Cardiff University, Cardiff, United Kingdom
Tracey A. Martin, Ph.D.
- Metastasis and Angiogenesis Research Group, Department of Surgery, School of Medicine, Cardiff University, Cardiff, United Kingdom
- Corresponding author. Tel.: +44-0-29-20744711; fax: +44-0-29-20761623.
Lin Ye, M.D.
- Metastasis and Angiogenesis Research Group, Department of Surgery, School of Medicine, Cardiff University, Cardiff, United Kingdom
Jonathon M. Lewis-Russell, M.D.
- Metastasis and Angiogenesis Research Group, Department of Surgery, School of Medicine, Cardiff University, Cardiff, United Kingdom
Malcolm D. Mason, M.D.
- Department of Medicine, Section of Clinical Oncology, Velindre Hospital, Cardiff and Vale NHS Trust, Cardiff, United Kingdom
Wen G. Jiang, M.D.
- Metastasis and Angiogenesis Research Group, Department of Surgery, School of Medicine, Cardiff University, Cardiff, United Kingdom

Received 22 February 2007; received in revised form 21 June 2007; accepted 21 June 2007. published online 15 January 2008.

Abstract

Background

Phospholipase C gamma-1 (PLCγ-1) is an intracellular signalling molecule regulating a number of biological processes including transporter mechanisms, transcription factors, and scaffolding proteins mediating cytoskeleton and membrane trafficking. Hepatocyte growth factor (HGF) is a pleiotrophic factor and a mediator of metastatic spread. This study sought to determine the effect of HGF on the invasive and migratory potential of prostate cancer cells targeted by a ribozyme transgene to PLCγ-1.

Methods

A ribozyme transgene consisting of hammerhead ribozyme and antisense specific to PLCγ-1 was cloned into a PEF6 expression vector and transfected into PC-3 cells. RT-PCR and Western blotting confirmed knock down of PLCγ-1. In vitro invasion and a cytodex-2 bead motility assays with in vitro and in vivo growth models were used to assess the impact of PLCγ-1 manipulation.

Results

PC-3 cells stably transfected with PLCγ-1 ribozyme transgene (PC-3^ΔPLCγ) manifested a reduction of PLCγ-1 expression at mRNA/protein levels. HGF/SF increased invasiveness (P < 0.01) and motility (P < 0.0001) of PC-3^WT and PC-3^PEF6 cells. In contrast, PLCγ-1 knock down PC-3^ΔPLCγ cells had reduced invasiveness (P < 0.05) and motility (P < 0.01) compared with PC-3^WT and PC-3^PEF6 cells. Although there was a marginal change of cell growth in vitro, there was no difference in the rate of growth between PC-3^ΔPLCγ, PC-3^WT, and PC-3^PEF6 cells.

Conclusion

Targeting PLCγ-1 by way of a hammerhead ribozyme to PLCγ-1 is an effective method in reducing invasive phenotype of prostate cancer. PLCγ-1 is a signalling intermediate that has prime influence on HGF induced cellular invasion and migration without affecting the growth of the cells.

Keywords: Ribozyme, HGF/SF, PLCγ-1, Prostate cancer, Migration, Invasion