The Role of the c-MET/HGF Signalling Axis as a Critical Regulator of Invasion and Migration in Colorectal Cancer

Conor Bradley, Philip Dunne, Keara Redmond, Darragh McArt, Ken Arthur, Stephen McQuaid, Jaine Blayney, Manuel Salto-Tellez, Patrick Johnston, Sandra Van Schaeybroeck

Research output: Chapter in Book/Report/Conference proceedingConference contribution

Abstract

Background
The c-MET proto-oncogene is frequently overexpressed (50-60%), amplified (5-10%), and mutated (5%) in colorectal cancer (CRC). Hepatocyte growth factor (HGF)-induced c-MET activation has been linked with enhanced proliferation, migration, survival and invasion, and has been suggested as a possible prognostic biomarker for CRC.

Method
To model CRC tumour cell invasion, we have generated invasive CRC subpopulations using Boyden Invasion chambers. Invasive cell lines were characterised for protein expression/activity by Western blotting, and analysed for migratory and invasive potential using the xCELLigence System. To model the CRC microenvironment, we have utilised a range of co-culture techniques with CRC cell lines and colon fibroblasts. c-MET expression in FFPE tissues was measured using IHC in a tissue microarray (TMA) derived from early stage CRC patients.

Results
HCT116 and LoVo invasive subpopulations showed a mesenchymal, migratory/invasive phenotype, and showed increased expression of the EMT marker SNAIL, CD44, and loss of E-Cadherin. In addition, increased expression/activation of c-MET was observed in these sublines. Inhibition of c-MET using RNAi abrogated both basal and HGF-induced migration/invasion in CRC cell lines. Co-culture of CRC cells with HGF expressing colon-derived fibroblasts leads to activation of c-MET, increasing the migratory/invasive capacity of the cancer cells. Significant increased expression levels of c-MET were also found in a CRC TMA compared to matched normal tissues. We are currently investigating the role of the tumour microenvironment in regulating migration/invasion and resistance to targeted therapy in CRC.

Conclusion
The identification of key pathways driving metastasis has a huge potential to change treatment strategies in CRC. We intend to further investigate the role of c-MET in regulation of migration and invasion in CRC in vitro and in vivo models. Characterisation of such novel mechanisms may provide a preclinical rationale for therapeutic c-MET inhibition in early stage CRC, specifically in patient subpopulations which display enhanced c-MET expression and activation.
Original languageEnglish
Title of host publicationThe Role of the c-MET/HGF Signalling Axis as a Critical Regulator of Invasion and Migration in Colorectal Cancer
PublisherNational Cancer Research Institute
Publication statusPublished - Nov 2014

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Hepatocyte Growth Factor
Colorectal Neoplasms
Tumor Microenvironment
Coculture Techniques
Cell Line
Colon
Fibroblasts
Culture Techniques
Proto-Oncogenes
Cadherins
RNA Interference

Cite this

Bradley, C., Dunne, P., Redmond, K., McArt, D., Arthur, K., McQuaid, S., ... Van Schaeybroeck, S. (2014). The Role of the c-MET/HGF Signalling Axis as a Critical Regulator of Invasion and Migration in Colorectal Cancer. In The Role of the c-MET/HGF Signalling Axis as a Critical Regulator of Invasion and Migration in Colorectal Cancer National Cancer Research Institute.
Bradley, Conor ; Dunne, Philip ; Redmond, Keara ; McArt, Darragh ; Arthur, Ken ; McQuaid, Stephen ; Blayney, Jaine ; Salto-Tellez, Manuel ; Johnston, Patrick ; Van Schaeybroeck, Sandra. / The Role of the c-MET/HGF Signalling Axis as a Critical Regulator of Invasion and Migration in Colorectal Cancer. The Role of the c-MET/HGF Signalling Axis as a Critical Regulator of Invasion and Migration in Colorectal Cancer. National Cancer Research Institute, 2014.
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title = "The Role of the c-MET/HGF Signalling Axis as a Critical Regulator of Invasion and Migration in Colorectal Cancer",
abstract = "BackgroundThe c-MET proto-oncogene is frequently overexpressed (50-60{\%}), amplified (5-10{\%}), and mutated (5{\%}) in colorectal cancer (CRC). Hepatocyte growth factor (HGF)-induced c-MET activation has been linked with enhanced proliferation, migration, survival and invasion, and has been suggested as a possible prognostic biomarker for CRC.MethodTo model CRC tumour cell invasion, we have generated invasive CRC subpopulations using Boyden Invasion chambers. Invasive cell lines were characterised for protein expression/activity by Western blotting, and analysed for migratory and invasive potential using the xCELLigence System. To model the CRC microenvironment, we have utilised a range of co-culture techniques with CRC cell lines and colon fibroblasts. c-MET expression in FFPE tissues was measured using IHC in a tissue microarray (TMA) derived from early stage CRC patients.ResultsHCT116 and LoVo invasive subpopulations showed a mesenchymal, migratory/invasive phenotype, and showed increased expression of the EMT marker SNAIL, CD44, and loss of E-Cadherin. In addition, increased expression/activation of c-MET was observed in these sublines. Inhibition of c-MET using RNAi abrogated both basal and HGF-induced migration/invasion in CRC cell lines. Co-culture of CRC cells with HGF expressing colon-derived fibroblasts leads to activation of c-MET, increasing the migratory/invasive capacity of the cancer cells. Significant increased expression levels of c-MET were also found in a CRC TMA compared to matched normal tissues. We are currently investigating the role of the tumour microenvironment in regulating migration/invasion and resistance to targeted therapy in CRC.ConclusionThe identification of key pathways driving metastasis has a huge potential to change treatment strategies in CRC. We intend to further investigate the role of c-MET in regulation of migration and invasion in CRC in vitro and in vivo models. Characterisation of such novel mechanisms may provide a preclinical rationale for therapeutic c-MET inhibition in early stage CRC, specifically in patient subpopulations which display enhanced c-MET expression and activation.",
author = "Conor Bradley and Philip Dunne and Keara Redmond and Darragh McArt and Ken Arthur and Stephen McQuaid and Jaine Blayney and Manuel Salto-Tellez and Patrick Johnston and {Van Schaeybroeck}, Sandra",
year = "2014",
month = "11",
language = "English",
booktitle = "The Role of the c-MET/HGF Signalling Axis as a Critical Regulator of Invasion and Migration in Colorectal Cancer",
publisher = "National Cancer Research Institute",

}

Bradley, C, Dunne, P, Redmond, K, McArt, D, Arthur, K, McQuaid, S, Blayney, J, Salto-Tellez, M, Johnston, P & Van Schaeybroeck, S 2014, The Role of the c-MET/HGF Signalling Axis as a Critical Regulator of Invasion and Migration in Colorectal Cancer. in The Role of the c-MET/HGF Signalling Axis as a Critical Regulator of Invasion and Migration in Colorectal Cancer. National Cancer Research Institute.

The Role of the c-MET/HGF Signalling Axis as a Critical Regulator of Invasion and Migration in Colorectal Cancer. / Bradley, Conor; Dunne, Philip; Redmond, Keara; McArt, Darragh; Arthur, Ken; McQuaid, Stephen; Blayney, Jaine; Salto-Tellez, Manuel; Johnston, Patrick; Van Schaeybroeck, Sandra.

The Role of the c-MET/HGF Signalling Axis as a Critical Regulator of Invasion and Migration in Colorectal Cancer. National Cancer Research Institute, 2014.

Research output: Chapter in Book/Report/Conference proceedingConference contribution

TY - GEN

T1 - The Role of the c-MET/HGF Signalling Axis as a Critical Regulator of Invasion and Migration in Colorectal Cancer

AU - Bradley, Conor

AU - Dunne, Philip

AU - Redmond, Keara

AU - McArt, Darragh

AU - Arthur, Ken

AU - McQuaid, Stephen

AU - Blayney, Jaine

AU - Salto-Tellez, Manuel

AU - Johnston, Patrick

AU - Van Schaeybroeck, Sandra

PY - 2014/11

Y1 - 2014/11

N2 - BackgroundThe c-MET proto-oncogene is frequently overexpressed (50-60%), amplified (5-10%), and mutated (5%) in colorectal cancer (CRC). Hepatocyte growth factor (HGF)-induced c-MET activation has been linked with enhanced proliferation, migration, survival and invasion, and has been suggested as a possible prognostic biomarker for CRC.MethodTo model CRC tumour cell invasion, we have generated invasive CRC subpopulations using Boyden Invasion chambers. Invasive cell lines were characterised for protein expression/activity by Western blotting, and analysed for migratory and invasive potential using the xCELLigence System. To model the CRC microenvironment, we have utilised a range of co-culture techniques with CRC cell lines and colon fibroblasts. c-MET expression in FFPE tissues was measured using IHC in a tissue microarray (TMA) derived from early stage CRC patients.ResultsHCT116 and LoVo invasive subpopulations showed a mesenchymal, migratory/invasive phenotype, and showed increased expression of the EMT marker SNAIL, CD44, and loss of E-Cadherin. In addition, increased expression/activation of c-MET was observed in these sublines. Inhibition of c-MET using RNAi abrogated both basal and HGF-induced migration/invasion in CRC cell lines. Co-culture of CRC cells with HGF expressing colon-derived fibroblasts leads to activation of c-MET, increasing the migratory/invasive capacity of the cancer cells. Significant increased expression levels of c-MET were also found in a CRC TMA compared to matched normal tissues. We are currently investigating the role of the tumour microenvironment in regulating migration/invasion and resistance to targeted therapy in CRC.ConclusionThe identification of key pathways driving metastasis has a huge potential to change treatment strategies in CRC. We intend to further investigate the role of c-MET in regulation of migration and invasion in CRC in vitro and in vivo models. Characterisation of such novel mechanisms may provide a preclinical rationale for therapeutic c-MET inhibition in early stage CRC, specifically in patient subpopulations which display enhanced c-MET expression and activation.

AB - BackgroundThe c-MET proto-oncogene is frequently overexpressed (50-60%), amplified (5-10%), and mutated (5%) in colorectal cancer (CRC). Hepatocyte growth factor (HGF)-induced c-MET activation has been linked with enhanced proliferation, migration, survival and invasion, and has been suggested as a possible prognostic biomarker for CRC.MethodTo model CRC tumour cell invasion, we have generated invasive CRC subpopulations using Boyden Invasion chambers. Invasive cell lines were characterised for protein expression/activity by Western blotting, and analysed for migratory and invasive potential using the xCELLigence System. To model the CRC microenvironment, we have utilised a range of co-culture techniques with CRC cell lines and colon fibroblasts. c-MET expression in FFPE tissues was measured using IHC in a tissue microarray (TMA) derived from early stage CRC patients.ResultsHCT116 and LoVo invasive subpopulations showed a mesenchymal, migratory/invasive phenotype, and showed increased expression of the EMT marker SNAIL, CD44, and loss of E-Cadherin. In addition, increased expression/activation of c-MET was observed in these sublines. Inhibition of c-MET using RNAi abrogated both basal and HGF-induced migration/invasion in CRC cell lines. Co-culture of CRC cells with HGF expressing colon-derived fibroblasts leads to activation of c-MET, increasing the migratory/invasive capacity of the cancer cells. Significant increased expression levels of c-MET were also found in a CRC TMA compared to matched normal tissues. We are currently investigating the role of the tumour microenvironment in regulating migration/invasion and resistance to targeted therapy in CRC.ConclusionThe identification of key pathways driving metastasis has a huge potential to change treatment strategies in CRC. We intend to further investigate the role of c-MET in regulation of migration and invasion in CRC in vitro and in vivo models. Characterisation of such novel mechanisms may provide a preclinical rationale for therapeutic c-MET inhibition in early stage CRC, specifically in patient subpopulations which display enhanced c-MET expression and activation.

M3 - Conference contribution

BT - The Role of the c-MET/HGF Signalling Axis as a Critical Regulator of Invasion and Migration in Colorectal Cancer

PB - National Cancer Research Institute

ER -

Bradley C, Dunne P, Redmond K, McArt D, Arthur K, McQuaid S et al. The Role of the c-MET/HGF Signalling Axis as a Critical Regulator of Invasion and Migration in Colorectal Cancer. In The Role of the c-MET/HGF Signalling Axis as a Critical Regulator of Invasion and Migration in Colorectal Cancer. National Cancer Research Institute. 2014