Title: Nuclear respiratory factor 1 promotes the growth of liver hepatocellular carcinoma cells
via E2F1 transcriptional activation
Version: 1 Date: 13 Sep 2021
Reviewer's report:
The manuscript entitled ”Nuclear respiratory factor 1 promotes the growth of liver hepatocellular
carcinoma cells via E2F1 transcriptional activation” is of valuble work done by Dan Wang et
al.  By combination of dry and wet experiments with clinical data, the authors analyzed the
relevance of NRF1 to  its occurrence of liver cancer. The effect of NRF1 on liver cancer was
also examined at the cellular level, so as ot gain insight into the relevant mechanism. The results
unravelled that NRF1 directly regulated the expression of  E2F1 as a cognate target gene, which
governs the proliferation of liver cancer by affecting its cell cycle process. Thereby, this is a
topic of interest to attract potential workers in all related areas. However, before being acceptable
for publication in this journal, this manuscript has to warrant better improvement to address the
following comments#
Firstly, the authors' biometric analysis lacks a comparison for their comprehensiveness and
verification, because they only used a single database or a single method. Why did not try
GEO data collection or TCGA to verify it. In the enrichment analysis, in addition to DAVID,
Clusterprofiler and other tools should also be used. Together, the author should conduct more
comprehensive data mining processing and verification analysis. The selected 192 NRF1-target
genes showed a correlation with the cell cycle by analyzing the ChIP-Seq data, but how to select
the core gene E2F1 should further be explained.
Secondly, the author examined how NRF1 affects the development of liver cancer, but with
a lack of a holistic discussion. The changes in the cell cycle were only briefly described after
the MTT experiment. No follow-up verification experiments, e.g., flow cytometry, cell scratch
and Transwell experiments were provided. Direct skip analysis concluded that NRF1 affects the
proliferation of liver cancer cells by regulating the cell cycle. In addition to the cellular level in
vitro, did the authors consider whether to use the mouse model to further explore the occurrence
of liver cancer with NRF1 in vivo?
Thirdly, NRF1 is closely related to the health and function of mitochondria, and dysfunction of
mitochondria directly affects the development of liver cancer and its malignant transformation.
Since the authors found that NRF1 regulates the cell cycle, they should examine potential effects
of NRF1 on mitochondrial function and genesis. The putative mechanistic findings should also
be discussed.
Title: Nuclear respiratory factor 1 promotes the growth of liver hepatocellular carcinoma cells
via E2F1 transcriptional activation
Version: 1 Date: 13 Sep 2021
Reviewer's report:
The manuscript entitled ”Nuclear respiratory factor 1 promotes the growth of liver hepatocellular
carcinoma cells via E2F1 transcriptional activation” is of valuble work done by Dan Wang et
al.  By combination of dry and wet experiments with clinical data, the authors analyzed the
relevance of NRF1 to  its occurrence of liver cancer. The effect of NRF1 on liver cancer was
also examined at the cellular level, so as ot gain insight into the relevant mechanism. The results
unravelled that NRF1 directly regulated the expression of  E2F1 as a cognate target gene, which
governs the proliferation of liver cancer by affecting its cell cycle process. Thereby, this is a
topic of interest to attract potential workers in all related areas. However, before being acceptable
for publication in this journal, this manuscript has to warrant better improvement to address the
following comments#
Firstly, the authors' biometric analysis lacks a comparison for their comprehensiveness and
verification, because they only used a single database or a single method. Why did not try
GEO data collection or TCGA to verify it. In the enrichment analysis, in addition to DAVID,
Clusterprofiler and other tools should also be used. Together, the author should conduct more
comprehensive data mining processing and verification analysis. The selected 192 NRF1-target
genes showed a correlation with the cell cycle by analyzing the ChIP-Seq data, but how to select
the core gene E2F1 should further be explained.
Secondly, the author examined how NRF1 affects the development of liver cancer, but with
a lack of a holistic discussion. The changes in the cell cycle were only briefly described after
the MTT experiment. No follow-up verification experiments, e.g., flow cytometry, cell scratch
and Transwell experiments were provided. Direct skip analysis concluded that NRF1 affects the
proliferation of liver cancer cells by regulating the cell cycle. In addition to the cellular level in
vitro, did the authors consider whether to use the mouse model to further explore the occurrence
of liver cancer with NRF1 in vivo?
Thirdly, NRF1 is closely related to the health and function of mitochondria, and dysfunction of
mitochondria directly affects the development of liver cancer and its malignant transformation.
Since the authors found that NRF1 regulates the cell cycle, they should examine potential effects
of NRF1 on mitochondrial function and genesis. The putative mechanistic findings should also
be discussed.
Please review this paper.
user
Title: Nuclear respiratory factor 1 promotes the growth of liver hepatocellular carcinoma cells
via E2F1 transcriptional activation
Version: 1 Date: 13 Sep 2021
Reviewer's report:
The manuscript entitled ”Nuclear respiratory factor 1 promotes the growth of liver hepatocellular
carcinoma cells via E2F1 transcriptional activation” is of valuble work done by Dan Wang et
al.  By combination of dry and wet experiments with clinical data, the authors analyzed the
relevance of NRF1 to  its occurrence of liver cancer. The effect of NRF1 on liver cancer was
also examined at the cellular level, so as ot gain insight into the relevant mechanism. The results
unravelled that NRF1 directly regulated the expression of  E2F1 as a cognate target gene, which
governs the proliferation of liver cancer by affecting its cell cycle process. Thereby, this is a
topic of interest to attract potential workers in all related areas. However, before being acceptable
for publication in this journal, this manuscript has to warrant better improvement to address the
following comments#
Firstly, the authors' biometric analysis lacks a comparison for their comprehensiveness and
verification, because they only used a single database or a single method. Why did not try
GEO data collection or TCGA to verify it. In the enrichment analysis, in addition to DAVID,
Clusterprofiler and other tools should also be used. Together, the author should conduct more
comprehensive data mining processing and verification analysis. The selected 192 NRF1-target
genes showed a correlation with the cell cycle by analyzing the ChIP-Seq data, but how to select
the core gene E2F1 should further be explained.
Secondly, the author examined how NRF1 affects the development of liver cancer, but with
a lack of a holistic discussion. The changes in the cell cycle were only briefly described after
the MTT experiment. No follow-up verification experiments, e.g., flow cytometry, cell scratch
and Transwell experiments were provided. Direct skip analysis concluded that NRF1 affects the
proliferation of liver cancer cells by regulating the cell cycle. In addition to the cellular level in
vitro, did the authors consider whether to use the mouse model to further explore the occurrence
of liver cancer with NRF1 in vivo?
Thirdly, NRF1 is closely related to the health and function of mitochondria, and dysfunction of
mitochondria directly affects the development of liver cancer and its malignant transformation.
Since the authors found that NRF1 regulates the cell cycle, they should examine potential effects
of NRF1 on mitochondrial function and genesis. The putative mechanistic findings should also
be discussed.
assistant