Reviewer's report
Title:Neoplastic transformation of rat liver epithelial cells is enhanced by
non-transferrin-bound iron.
Version: 1Date:5 September 2007
Reviewer: Yutaka Kohgo
Reviewer's report:
General
Major Compulsory Revisions (that the author must respond to before a decision
on publication can be reached)
This manuscript describes that iron loading by ferric ammonium citrate (FAC) to
the T51B cells (liver epithelial cells) function as a tumor promoter, and cell cycle
dysregulation might contribute to tumor promotion by FAC. The results presented
in this report are interesting because the authors investigated the transforming
effects of FAC, which is one of the non-transferrin-bound iron (NTBI) observed in
iron overload, and this might be the first report focusing on this point. However, a
few criticisms and unclear points may be raised on the present form of this paper.
1. The authors used T51B cell line that was derived from hepatic epithelial cells.
The authors must comment if the result from the experiments using this cell line
directly proves that the increase of hepatoma in iron overload, because
hepatoma is derived from hepatocytes, not hepatic epithelial cells.
2. The authors should comment the physiological possible concentration of FAC,
especially in the situation of iron overload state. It should be important to tell us
whether the concentration of FAC around 200#M is actually observed
concentration in human. If the concentrations of FAC were much higher than
those observed in human with iron overload, the results should be considered
carefully.
3. What kind of uptake route did the T51B cell line use to take up FAC from the
medium? The authors should comment about that.
4. The authors investigated the iron status after iron loading with FAC in T51B
cells. However, they presented the results only for 5 days incubation. They
incubated T51B cells for more than 12 weeks in other experiments, so that they
should show the result of iron status of T51B cells that were incubated for more
than 12 weeks.
5. The incubation times and periods in there experiments are different. For
example, the authors cultured T51B cells for 12 weeks in Table 1, but they
cultured the cells for 16 weeks in Figure 3. The authors should tell us how they
settled the incubation times in each experiment in the “materials and methods”.
6. The authors should comment why they chose FAC as the model of NTBI in
their experiments. There should be many kinds of NTBI molecules in iron
overload situation, so that it should be better to tell the reasons why the authors
selected FAC.
----------------------------------------------------------------------------
Minor Essential Revisions (such as missing labels on figures, or the wrong use of
a term, which the author can be trusted to correct)
1.It should be better to write full-spelling of TPA, before using abbreviation. (In
Discussion, 2nd paragraph, line2)
-------------------------------------------------------------------------------
Discretionary Revisions (which the author can choose to ignore)
What next?: Unable to decide on acceptance or rejection until the authors have
responded to the major compulsory revisions
Level of interest: An article of importance in its field
Quality of written English: Acceptable
Statistical review: No, the manuscript does not need to be seen by a
statistician.
Declaration of competing interests:
'I declare that I have no competing interests'
Reviewer's report
Title:Neoplastic transformation of rat liver epithelial cells is enhanced by
non-transferrin-bound iron.
Version: 1Date:5 September 2007
Reviewer: Yutaka Kohgo
Reviewer's report:
General
Major Compulsory Revisions (that the author must respond to before a decision
on publication can be reached)
This manuscript describes that iron loading by ferric ammonium citrate (FAC) to
the T51B cells (liver epithelial cells) function as a tumor promoter, and cell cycle
dysregulation might contribute to tumor promotion by FAC. The results presented
in this report are interesting because the authors investigated the transforming
effects of FAC, which is one of the non-transferrin-bound iron (NTBI) observed in
iron overload, and this might be the first report focusing on this point. However, a
few criticisms and unclear points may be raised on the present form of this paper.
1. The authors used T51B cell line that was derived from hepatic epithelial cells.
The authors must comment if the result from the experiments using this cell line
directly proves that the increase of hepatoma in iron overload, because
hepatoma is derived from hepatocytes, not hepatic epithelial cells.
2. The authors should comment the physiological possible concentration of FAC,
especially in the situation of iron overload state. It should be important to tell us
whether the concentration of FAC around 200#M is actually observed
concentration in human. If the concentrations of FAC were much higher than
those observed in human with iron overload, the results should be considered
carefully.
3. What kind of uptake route did the T51B cell line use to take up FAC from the
medium? The authors should comment about that.
4. The authors investigated the iron status after iron loading with FAC in T51B
cells. However, they presented the results only for 5 days incubation. They
incubated T51B cells for more than 12 weeks in other experiments, so that they
should show the result of iron status of T51B cells that were incubated for more
than 12 weeks.
5. The incubation times and periods in there experiments are different. For
example, the authors cultured T51B cells for 12 weeks in Table 1, but they
cultured the cells for 16 weeks in Figure 3. The authors should tell us how they
settled the incubation times in each experiment in the “materials and methods”.
6. The authors should comment why they chose FAC as the model of NTBI in
their experiments. There should be many kinds of NTBI molecules in iron
overload situation, so that it should be better to tell the reasons why the authors
selected FAC.
----------------------------------------------------------------------------
Minor Essential Revisions (such as missing labels on figures, or the wrong use of
a term, which the author can be trusted to correct)
1.It should be better to write full-spelling of TPA, before using abbreviation. (In
Discussion, 2nd paragraph, line2)
-------------------------------------------------------------------------------
Discretionary Revisions (which the author can choose to ignore)
What next?: Unable to decide on acceptance or rejection until the authors have
responded to the major compulsory revisions
Level of interest: An article of importance in its field
Quality of written English: Acceptable
Statistical review: No, the manuscript does not need to be seen by a
statistician.
Declaration of competing interests:
'I declare that I have no competing interests'
Please review this paper.
user
Reviewer's report
Title:Neoplastic transformation of rat liver epithelial cells is enhanced by
non-transferrin-bound iron.
Version: 1Date:5 September 2007
Reviewer: Yutaka Kohgo
Reviewer's report:
General
Major Compulsory Revisions (that the author must respond to before a decision
on publication can be reached)
This manuscript describes that iron loading by ferric ammonium citrate (FAC) to
the T51B cells (liver epithelial cells) function as a tumor promoter, and cell cycle
dysregulation might contribute to tumor promotion by FAC. The results presented
in this report are interesting because the authors investigated the transforming
effects of FAC, which is one of the non-transferrin-bound iron (NTBI) observed in
iron overload, and this might be the first report focusing on this point. However, a
few criticisms and unclear points may be raised on the present form of this paper.
1. The authors used T51B cell line that was derived from hepatic epithelial cells.
The authors must comment if the result from the experiments using this cell line
directly proves that the increase of hepatoma in iron overload, because
hepatoma is derived from hepatocytes, not hepatic epithelial cells.
2. The authors should comment the physiological possible concentration of FAC,
especially in the situation of iron overload state. It should be important to tell us
whether the concentration of FAC around 200#M is actually observed
concentration in human. If the concentrations of FAC were much higher than
those observed in human with iron overload, the results should be considered
carefully.
3. What kind of uptake route did the T51B cell line use to take up FAC from the
medium? The authors should comment about that.
4. The authors investigated the iron status after iron loading with FAC in T51B
cells. However, they presented the results only for 5 days incubation. They
incubated T51B cells for more than 12 weeks in other experiments, so that they
should show the result of iron status of T51B cells that were incubated for more
than 12 weeks.
5. The incubation times and periods in there experiments are different. For
example, the authors cultured T51B cells for 12 weeks in Table 1, but they
cultured the cells for 16 weeks in Figure 3. The authors should tell us how they
settled the incubation times in each experiment in the “materials and methods”.
6. The authors should comment why they chose FAC as the model of NTBI in
their experiments. There should be many kinds of NTBI molecules in iron
overload situation, so that it should be better to tell the reasons why the authors
selected FAC.
----------------------------------------------------------------------------
Minor Essential Revisions (such as missing labels on figures, or the wrong use of
a term, which the author can be trusted to correct)
1.It should be better to write full-spelling of TPA, before using abbreviation. (In
Discussion, 2nd paragraph, line2)
-------------------------------------------------------------------------------
Discretionary Revisions (which the author can choose to ignore)
What next?: Unable to decide on acceptance or rejection until the authors have
responded to the major compulsory revisions
Level of interest: An article of importance in its field
Quality of written English: Acceptable
Statistical review: No, the manuscript does not need to be seen by a
statistician.
Declaration of competing interests:
'I declare that I have no competing interests'
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