Reviewer's report 
 Epidermal growth factor mediates detachment from and invasion through collagen I and Title: 
Matrigel in Capan-1 pancreatic cancer cells 
 1  24 October 2004 Version: Date: 
 Jorg Kleeff Reviewer: 
Reviewer's report: 
'Epidermal growth factor mediates detachment from and invasion through collagen I and Matrigel in 
Capan-1 pancreatic cancer cells ' BMC Gastroenterology 
The study by Shirk and Kuver examines the role of EGF on invasion and adhesion in Capan-1 
pancreatic cancer cells. The authors demonstrate that there is an inverse relationship between the invasion and adhesions capabilities and that these events are mediated by EGF receptor signalling involving the erbB2 and PI3K pathways. 
This is an interesting study and the experimental procedures are in general sound and valid. 
Although most of the aspects of the present study are not completely novel and have been 
described in other cell system, the present paper adds some novel information regarding the role of 
EGF in pancreatic cancer. There are, however, certain points that should be addressed: 
Major Compulsory Revisions 
• The authors utilized only a single pancreatic cancer cell line (Capan-1) to analyze the effects of EGF on invasion and adhesion. It would be important to use at least one additional pancreatic 
cancer cell line and perform some of the key experiments in this cell line. 
• The authors should provide information regarding the expression levels of all (!) four EGF receptors 
in the analyzed cell lines. This information is crucial especially regarding the final conclusion of the 
present paper. It would be interesting to perform this analysis quantitatively (e.g. immunoblotting). 
• The authors utilized EGF and for some additional experiments also heregulin-alpha. The rationale 
for using this (and not other) growth factors of the EGF family is not clear and should be elaborated. • The authors speculate that EGF acted via erbB1 and erbB2 and heregulin via erbB3 and erbB4. However, the authors do not provide any good experimental evidence for this conclusion. The 
authors should either demonstrate for example the activation/phosphorylation of the respective 
receptors in response to the growth factors or analyze the effects of specific blockage of these receptors. • In this context, for further experiments, the authors utilized erbB2 and erbB3 inhibitors /receptor 
antibodies. The authors should also analyze EGFR inhibitors /receptor antibodies for these 
experiments. 
 Unable to decide on acceptance or rejection until the authors have responded to the What next?: 
major compulsory revisions 
 An article whose findings are important to those with closely related research Level of interest: 
interests 
 Acceptable Quality of written English: 
 No Statistical review: 
Declaration of competing interests: 
None 
Reviewer's report 
 Epidermal growth factor mediates detachment from and invasion through collagen I and Title: 
Matrigel in Capan-1 pancreatic cancer cells 
 1  24 October 2004 Version: Date: 
 Jorg Kleeff Reviewer: 
Reviewer's report: 
'Epidermal growth factor mediates detachment from and invasion through collagen I and Matrigel in 
Capan-1 pancreatic cancer cells ' BMC Gastroenterology 
The study by Shirk and Kuver examines the role of EGF on invasion and adhesion in Capan-1 
pancreatic cancer cells. The authors demonstrate that there is an inverse relationship between the invasion and adhesions capabilities and that these events are mediated by EGF receptor signalling involving the erbB2 and PI3K pathways. 
This is an interesting study and the experimental procedures are in general sound and valid. 
Although most of the aspects of the present study are not completely novel and have been 
described in other cell system, the present paper adds some novel information regarding the role of 
EGF in pancreatic cancer. There are, however, certain points that should be addressed: 
Major Compulsory Revisions 
• The authors utilized only a single pancreatic cancer cell line (Capan-1) to analyze the effects of EGF on invasion and adhesion. It would be important to use at least one additional pancreatic 
cancer cell line and perform some of the key experiments in this cell line. 
• The authors should provide information regarding the expression levels of all (!) four EGF receptors 
in the analyzed cell lines. This information is crucial especially regarding the final conclusion of the 
present paper. It would be interesting to perform this analysis quantitatively (e.g. immunoblotting). 
• The authors utilized EGF and for some additional experiments also heregulin-alpha. The rationale 
for using this (and not other) growth factors of the EGF family is not clear and should be elaborated. • The authors speculate that EGF acted via erbB1 and erbB2 and heregulin via erbB3 and erbB4. However, the authors do not provide any good experimental evidence for this conclusion. The 
authors should either demonstrate for example the activation/phosphorylation of the respective 
receptors in response to the growth factors or analyze the effects of specific blockage of these receptors. • In this context, for further experiments, the authors utilized erbB2 and erbB3 inhibitors /receptor 
antibodies. The authors should also analyze EGFR inhibitors /receptor antibodies for these 
experiments. 
 Unable to decide on acceptance or rejection until the authors have responded to the What next?: 
major compulsory revisions 
 An article whose findings are important to those with closely related research Level of interest: 
interests 
 Acceptable Quality of written English: 
 No Statistical review: 
Declaration of competing interests: 
None 
Please review this paper.
user
Reviewer's report 
 Epidermal growth factor mediates detachment from and invasion through collagen I and Title: 
Matrigel in Capan-1 pancreatic cancer cells 
 1  24 October 2004 Version: Date: 
 Jorg Kleeff Reviewer: 
Reviewer's report: 
'Epidermal growth factor mediates detachment from and invasion through collagen I and Matrigel in 
Capan-1 pancreatic cancer cells ' BMC Gastroenterology 
The study by Shirk and Kuver examines the role of EGF on invasion and adhesion in Capan-1 
pancreatic cancer cells. The authors demonstrate that there is an inverse relationship between the invasion and adhesions capabilities and that these events are mediated by EGF receptor signalling involving the erbB2 and PI3K pathways. 
This is an interesting study and the experimental procedures are in general sound and valid. 
Although most of the aspects of the present study are not completely novel and have been 
described in other cell system, the present paper adds some novel information regarding the role of 
EGF in pancreatic cancer. There are, however, certain points that should be addressed: 
Major Compulsory Revisions 
• The authors utilized only a single pancreatic cancer cell line (Capan-1) to analyze the effects of EGF on invasion and adhesion. It would be important to use at least one additional pancreatic 
cancer cell line and perform some of the key experiments in this cell line. 
• The authors should provide information regarding the expression levels of all (!) four EGF receptors 
in the analyzed cell lines. This information is crucial especially regarding the final conclusion of the 
present paper. It would be interesting to perform this analysis quantitatively (e.g. immunoblotting). 
• The authors utilized EGF and for some additional experiments also heregulin-alpha. The rationale 
for using this (and not other) growth factors of the EGF family is not clear and should be elaborated. • The authors speculate that EGF acted via erbB1 and erbB2 and heregulin via erbB3 and erbB4. However, the authors do not provide any good experimental evidence for this conclusion. The 
authors should either demonstrate for example the activation/phosphorylation of the respective 
receptors in response to the growth factors or analyze the effects of specific blockage of these receptors. • In this context, for further experiments, the authors utilized erbB2 and erbB3 inhibitors /receptor 
antibodies. The authors should also analyze EGFR inhibitors /receptor antibodies for these 
experiments. 
 Unable to decide on acceptance or rejection until the authors have responded to the What next?: 
major compulsory revisions 
 An article whose findings are important to those with closely related research Level of interest: 
interests 
 Acceptable Quality of written English: 
 No Statistical review: 
Declaration of competing interests: 
None 
assistant