Title: Expression of kinetochore component NDC80 promotes esophageal squamous cell
carcinoma cells proliferation and migration
Version: 1 Date: 04 Jan 2024
Reviewer:  Mehdi Montazer
Reviewer's report:
The authors have investigated the expression of NDC80, a kinetochore gene, in esophageal
squamous cell carcinoma cells (ESCC) and evaluated its role in proliferation and invasive
characteristics of ESCC cells using a variety of in vitro methods, including colony formation,
apoptosis assessment, wound-healing, and transwell migration. It has been previously
demonstrated through computational analysis that NDC80 plays a predictive role. However, as
far as I know, this is the first instance where the relevant mechanisms have been experimentally
verified. The chosen techniques are suitable, and the results are of interest to experts in this
particular area.
Major comments:
1. The manuscript needs to undergo English editing.
2. It is already known from computational studies that a high expression of NDC80 is
associated with a worse prognosis (Yang et al. Cancer Cell Int (2019) 19:142, https://
doi.org/10.1186/s12935-019-0854-6; Xiao Wang et al. https://doi.org/10.1155/2022/3758731).
It is recommended to mention these computational findings and the relevant references in the
introduction and discussion parts and concentrate instead on the idea that the present study aimed
to confirm and decipher such findings experimentally. In fact, I do not agree with statements like
“the role of NDC80 in esophageal squamous cell carcinoma (ESCC) is still unknown”, “Further
studies with larger sample size are needed to confirm its influence in ESCC survival”, and “But
the impact of NDC80 expression on esophageal cancer was still unknown”. The overall role is
already known.
3. Sequencing of NDC80 and Data Analyses: 1. What did the authors use normal tissues
(including 43 blood samples) for? Did they use them to omit any possible germline variants?
2. Please provide the primary metrics of the NGS quality, such as mean coverage and mapping
quality. 3. Including BAM file images showing NDC80 gain can enrich the paper.
4. It looks like the shRNA sequence and real-time PCR primers are taken from previously
published articles (Am J Cancer Res 2015;5(5):1730-1740 www.ajcr.us /ISSN:2156-6976/
ajcr0006976 and World J Gastroenterol. 2017 May 28; 23(20): 3675–3683. doi: 10.3748/
wjg.v23.i20.3675). Please cite these papers.
5. Line 180, “Suspended cells were wash away and cells were cultured in DMEM medium with
0.5% FSB at 37°C”. Why did the authors culture these cells (scraped cells?)?
6. The reads are aligned with the hg19 genome assembly while SNVs are reported based on the
GRCh38.p13 (hg38). Please confirm that you have converted the genome coordinates.
7. Please include the mean +/- SE OS for normal and neutral copy number patients. They are
now grouped in Figure 1 and I am curious to know their status separately. What is more, how
many patients had a normal, copy neutral or complete loss?
8. Please use the standard nomenclature (HGVS format, https://varnomen.hgvs.org/) for
reporting the novel SNP.
9. Statistical analysis: 1. The authors have used parametric tests to make comparisons between
means. Were the variables normally distributed? To have a normal distribution is a constant
assumption of such tests. 2. Where did the authors use the ANOVA test?
Minor comments:
1. Line 146: I suggest moving “ECA109 and TE1 cells were cultured in DMEM media and
transfected by lentivirus” to “shRNA interference of NDC80 and infections” part.
2. Please state what levels of p-value are represented by * and ** throughout the paper.
3. The statistics (mean +/- SE) for shRNA and Ctrl groups are written in place of each other on
several occasions: lines 232, 245, 261-263, 271-272, 277, 279, and 283-284.
4. Lines 165 and 185: Please correct the CO2.
5. Line 326: Please replace the link with the word “PubMed”.
Title: Expression of kinetochore component NDC80 promotes esophageal squamous cell
carcinoma cells proliferation and migration
Version: 1 Date: 04 Jan 2024
Reviewer:  Mehdi Montazer
Reviewer's report:
The authors have investigated the expression of NDC80, a kinetochore gene, in esophageal
squamous cell carcinoma cells (ESCC) and evaluated its role in proliferation and invasive
characteristics of ESCC cells using a variety of in vitro methods, including colony formation,
apoptosis assessment, wound-healing, and transwell migration. It has been previously
demonstrated through computational analysis that NDC80 plays a predictive role. However, as
far as I know, this is the first instance where the relevant mechanisms have been experimentally
verified. The chosen techniques are suitable, and the results are of interest to experts in this
particular area.
Major comments:
1. The manuscript needs to undergo English editing.
2. It is already known from computational studies that a high expression of NDC80 is
associated with a worse prognosis (Yang et al. Cancer Cell Int (2019) 19:142, https://
doi.org/10.1186/s12935-019-0854-6; Xiao Wang et al. https://doi.org/10.1155/2022/3758731).
It is recommended to mention these computational findings and the relevant references in the
introduction and discussion parts and concentrate instead on the idea that the present study aimed
to confirm and decipher such findings experimentally. In fact, I do not agree with statements like
“the role of NDC80 in esophageal squamous cell carcinoma (ESCC) is still unknown”, “Further
studies with larger sample size are needed to confirm its influence in ESCC survival”, and “But
the impact of NDC80 expression on esophageal cancer was still unknown”. The overall role is
already known.
3. Sequencing of NDC80 and Data Analyses: 1. What did the authors use normal tissues
(including 43 blood samples) for? Did they use them to omit any possible germline variants?
2. Please provide the primary metrics of the NGS quality, such as mean coverage and mapping
quality. 3. Including BAM file images showing NDC80 gain can enrich the paper.
4. It looks like the shRNA sequence and real-time PCR primers are taken from previously
published articles (Am J Cancer Res 2015;5(5):1730-1740 www.ajcr.us /ISSN:2156-6976/
ajcr0006976 and World J Gastroenterol. 2017 May 28; 23(20): 3675–3683. doi: 10.3748/
wjg.v23.i20.3675). Please cite these papers.
5. Line 180, “Suspended cells were wash away and cells were cultured in DMEM medium with
0.5% FSB at 37°C”. Why did the authors culture these cells (scraped cells?)?
6. The reads are aligned with the hg19 genome assembly while SNVs are reported based on the
GRCh38.p13 (hg38). Please confirm that you have converted the genome coordinates.
7. Please include the mean +/- SE OS for normal and neutral copy number patients. They are
now grouped in Figure 1 and I am curious to know their status separately. What is more, how
many patients had a normal, copy neutral or complete loss?
8. Please use the standard nomenclature (HGVS format, https://varnomen.hgvs.org/) for
reporting the novel SNP.
9. Statistical analysis: 1. The authors have used parametric tests to make comparisons between
means. Were the variables normally distributed? To have a normal distribution is a constant
assumption of such tests. 2. Where did the authors use the ANOVA test?
Minor comments:
1. Line 146: I suggest moving “ECA109 and TE1 cells were cultured in DMEM media and
transfected by lentivirus” to “shRNA interference of NDC80 and infections” part.
2. Please state what levels of p-value are represented by * and ** throughout the paper.
3. The statistics (mean +/- SE) for shRNA and Ctrl groups are written in place of each other on
several occasions: lines 232, 245, 261-263, 271-272, 277, 279, and 283-284.
4. Lines 165 and 185: Please correct the CO2.
5. Line 326: Please replace the link with the word “PubMed”.
Please review this paper.
user
Title: Expression of kinetochore component NDC80 promotes esophageal squamous cell
carcinoma cells proliferation and migration
Version: 1 Date: 04 Jan 2024
Reviewer:  Mehdi Montazer
Reviewer's report:
The authors have investigated the expression of NDC80, a kinetochore gene, in esophageal
squamous cell carcinoma cells (ESCC) and evaluated its role in proliferation and invasive
characteristics of ESCC cells using a variety of in vitro methods, including colony formation,
apoptosis assessment, wound-healing, and transwell migration. It has been previously
demonstrated through computational analysis that NDC80 plays a predictive role. However, as
far as I know, this is the first instance where the relevant mechanisms have been experimentally
verified. The chosen techniques are suitable, and the results are of interest to experts in this
particular area.
Major comments:
1. The manuscript needs to undergo English editing.
2. It is already known from computational studies that a high expression of NDC80 is
associated with a worse prognosis (Yang et al. Cancer Cell Int (2019) 19:142, https://
doi.org/10.1186/s12935-019-0854-6; Xiao Wang et al. https://doi.org/10.1155/2022/3758731).
It is recommended to mention these computational findings and the relevant references in the
introduction and discussion parts and concentrate instead on the idea that the present study aimed
to confirm and decipher such findings experimentally. In fact, I do not agree with statements like
“the role of NDC80 in esophageal squamous cell carcinoma (ESCC) is still unknown”, “Further
studies with larger sample size are needed to confirm its influence in ESCC survival”, and “But
the impact of NDC80 expression on esophageal cancer was still unknown”. The overall role is
already known.
3. Sequencing of NDC80 and Data Analyses: 1. What did the authors use normal tissues
(including 43 blood samples) for? Did they use them to omit any possible germline variants?
2. Please provide the primary metrics of the NGS quality, such as mean coverage and mapping
quality. 3. Including BAM file images showing NDC80 gain can enrich the paper.
4. It looks like the shRNA sequence and real-time PCR primers are taken from previously
published articles (Am J Cancer Res 2015;5(5):1730-1740 www.ajcr.us /ISSN:2156-6976/
ajcr0006976 and World J Gastroenterol. 2017 May 28; 23(20): 3675–3683. doi: 10.3748/
wjg.v23.i20.3675). Please cite these papers.
5. Line 180, “Suspended cells were wash away and cells were cultured in DMEM medium with
0.5% FSB at 37°C”. Why did the authors culture these cells (scraped cells?)?
6. The reads are aligned with the hg19 genome assembly while SNVs are reported based on the
GRCh38.p13 (hg38). Please confirm that you have converted the genome coordinates.
7. Please include the mean +/- SE OS for normal and neutral copy number patients. They are
now grouped in Figure 1 and I am curious to know their status separately. What is more, how
many patients had a normal, copy neutral or complete loss?
8. Please use the standard nomenclature (HGVS format, https://varnomen.hgvs.org/) for
reporting the novel SNP.
9. Statistical analysis: 1. The authors have used parametric tests to make comparisons between
means. Were the variables normally distributed? To have a normal distribution is a constant
assumption of such tests. 2. Where did the authors use the ANOVA test?
Minor comments:
1. Line 146: I suggest moving “ECA109 and TE1 cells were cultured in DMEM media and
transfected by lentivirus” to “shRNA interference of NDC80 and infections” part.
2. Please state what levels of p-value are represented by * and ** throughout the paper.
3. The statistics (mean +/- SE) for shRNA and Ctrl groups are written in place of each other on
several occasions: lines 232, 245, 261-263, 271-272, 277, 279, and 283-284.
4. Lines 165 and 185: Please correct the CO2.
5. Line 326: Please replace the link with the word “PubMed”.
assistant