Verified Rigor RecordRigor Reviewed

Rigor-Reviewed Manuscript

circRNA_100290 plays a role in oral cancer by functioning as a sponge of the miR-29 family · 10.1038/onc.2017.89

Name: Verified Rigor Record — circRNA_100290 plays a role in oral cancer by functioning as a sponge of the miR-29 family · 10.1038/onc.2017.89
Item: circRNA_100290 plays a role in oral cancer by functioning as a sponge of the miR-29 family · 10.1038/onc.2017.89
Rating: 67
Author: Αlpha¹

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Αlpha¹ Editorial

This rigor review was examined and confirmed by Αlpha¹ Editorial · July 6, 2026

Αlpha¹ Rigor Review

Rigor Reviewed

WARN

Warnings to address · Observational Study

Eight dimensions — click a segment for details

Critical

High-priority

Model-reported confidence: moderateReviewer agreement: highEngine: 7.6.0Source: full text

Not verified in this report

No reported statistical tests were found to recompute.

This Αlpha¹ Rigor Review uses AI Rigor Reviewers trained on a curated corpus of high-fidelity and retracted papers, with expert supervision and curation. It can still make mistakes; verify each finding against the source before relying on it.

The paper presents a logical scientific premise and uses appropriate controls, but is undermined by multiple reporting gaps: no data/code availability, missing ethics details for the animal study, and absent randomization/blinding/power analysis. The statistical reporting is imprecise (threshold p-values only) and key biological variables (patient demographics, cell line authentication) are not reported.

All three independent reviewers converged on the status for 7 of 8 dimensions; the only divergence was on ethical approvals (one reviewer rated 'pass' due to the IRB statement, but the missing IACUC and informed consent led to a synthesized 'warn'). The statistics verification component found no tests to recompute (all p-values were threshold-only), so no mathematical errors were identified, but this also means the reported statistics are unverified.

12 major claims checked against the paper's own evidence: all adequately supported.

1 integrity concern flagged (0 high).

Rigor

Eight Dimensions1 pass · 6 warn · 1 fail

12 major claims1 partial11 supported

The introduction cites numerous studies on miRNAs and lncRNAs in OSCC and the ceRNA hypothesis, establishing the premise that circRNAs may function as miRNA sponges. The rationale linking circRNA_100290 to CDK6 via the miR-29 family is logically derived from microarray and bioinformatic analyses. Limitations of prior research (i.e., few circRNAs confirmed as miRNA sponges) are acknowledged, and the gap in OSCC is explicitly stated.

Sub-criteria

prior work citedADEQUATE

premise rationaleADEQUATE

limitations addressedADEQUATE

Per reviewer (3) — the votes behind the verdict

PASSReviewer 1· 95% conf

The study clearly establishes the scientific foundation for investigating circRNAs in OSCC, citing relevant literature on non-coding RNAs and the ceRNA hypothesis, and identifies the gap in knowledge about circRNAs in OSCC.

Evidence

direct quote[Introduction, paragraphs 2-4]

“Over the past decade, microRNAs (miRNAs) and long non-coding RNAs (lncRNAs) have been found to serve as important players in a variety of physiology and pathological processes.”

paraphrase[Results, ceRNA analysis section]

“Paraphrase: The paper constructs a circRNA_100290-miRNA-target gene network from microarray data, identifying miR-29 family members and CDK6 as top predicted targets.”

direct quote[Discussion, paragraph 1]

“However, whether circRNAs play a role in OSCC is unknown. This is the first report on the expression profile and regulatory function of circRNAs in OSCC.”

PASSReviewer 2· 90% conf

The paper establishes a clear scientific premise by citing prior work on circRNAs, miRNAs, and the ceRNA hypothesis, and provides a logical rationale for investigating circRNA_100290 in OSCC.

Evidence

direct quote[Introduction, paragraphs 1-5]

“Pandolfi et al., presented a hypothesis called the competing endogenous RNA (ceRNA) mechanism, which proposed that transcripts such as mRNAs, pseudogenes and lncRNAs can serve as natural miRNA sponges by competitive binding to miRNA response elements (MREs) to suppress their expression and function.”

direct quote[Introduction, paragraph 5]

“In this study, we analysed the expression profiles of circRNAs and mRNAs in OSCC tissue through microarrays.”

direct quote[Introduction, paragraph 4]

“However, the function of circRNAs in tumour initiation and progression is unclear. Moreover, there are currently no reports describing the role of circRNAs in OSCC.”

PASSReviewer 3· 95% conf

The paper establishes the scientific premise for circRNA_100290 in OSCC, citing prior work on circRNAs and ceRNA mechanisms, and explains the rationale for the study.

Evidence

direct quote[Introduction, paragraph 5]

“only three circRNAs, ciRs-7 (also termed CDR1as), Sry and circHIPK3 have thus far been confirmed to function through a ceRNA mechanism”

direct quote[Introduction, paragraph 4]

“However, there are currently no reports describing the role of circRNAs in OSCC.”

direct quote[Introduction, paragraph 4]

“However, there are very few such circRNAs that contain multiple MREs to decoy a particular miRNA.”

The study is a mixed observational (case-control expression profiling) and interventional (cell line and xenograft) design. For the interventional components, randomization is not mentioned. Blinding of investigators or assessors is not reported. No power analysis is performed. The distinction between biological and technical replicates is unclear (figures state 'three separate experiments' but the level of replication for the in vivo experiment is minimal: 3 mice per group). Inclusion/exclusion criteria for patient samples and cell lines are not pre-specified. Outlier handling is not discussed. Controls are adequate for the functional assays (siRNA targeting linear transcript, miR mimics/inhibitors, negative controls). Independent replication of key findings is not reported.

Sub-criteria

controlsADEQUATE

power analysisNOT REPORTED

blinding levelsNOT REPORTED

outlier handlingNOT REPORTED

randomization unitNOT REPORTED

inclusion exclusionNOT REPORTED

randomization methodNOT REPORTED

replicate distinctionINADEQUATE

independent replicationNOT REPORTED

Per reviewer (3) — the votes behind the verdict

WARNReviewer 1· 85% conf

The study design has several strengths (use of multiple cell lines, in vivo model, multiple assays) but lacks critical elements: no randomization method, no blinding, no power analysis, no pre-specified inclusion/exclusion criteria, no outlier handling, and the biological vs. technical replicate distinction is unclear. Controls are present (siRNAs targeting linear transcript, miR inhibitors, negative controls) but not systematically described.

Evidence

absence[Methods, all sections]

Randomization method and unit are not mentioned.

absence[Methods, all sections]

Blinding of investigators or assessors is not reported.

direct quote[Methods, RNA interference and transfection assay]

“Small interfering RNAs (siRNAs) targeting the back-splice junction of circRNA_100290 (si-circRNA_100290) and siRNAs targeting the SLC30A7 linear transcript only (si-SLC30A7) were designed and synthesized by RiboBio (Guangzhou, China; ).”

WARNReviewer 2· 85% conf

The study design is partially rigorous, with adequate controls and replication, but lacks key elements like randomization, blinding, and a pre-specified power analysis.

Evidence

direct quote[Materials and methods, Tissue specimens]

“All of the patients enrolled in this study had not undergone previous surgery, radiotherapy and chemotherapy.”

direct quote[Results, Knockdown of circRNA_100290 inhibits expression of CDK6]

“To prove whether circRNA_100290 functions as a ceRNA, we co-transfected inhibitors of the miR-29 family and siRNA against circRNA_100290 into CAL27 and HN4 cell lines, and observed that the repression was reversed.”

direct quote[Results, Effects of circRNA_100290 on cell cycle and cell proliferation]

“To confirm these findings in in vivo models, HN4 cell lines were implanted subcutaneously into the pectoral region of nude mice (three mice for negative control group, three mice for si-circRNA_100290 group and three mice for si-circRNA_100290+miR-29 inhibitor group).”

WARNReviewer 3· 85% conf

The study design includes key elements like controls and siRNA specificity but lacks pre-specified inclusion/exclusion criteria for subjects, a power analysis, and randomization/blinding for the animal experiment.

Evidence

absence[Materials and methods, and Results]

No power analysis for the study.

direct quote[Materials and methods, Tissue specimens]

“Tissue samples of OSCC and paired NCMT were obtained from patients who received surgical treatment at the Second Xiangya Hospital”

direct quote[Results, paragraph 10]

“three mice for negative control group, three mice for si-circRNA_100290 group and three mice for si-circRNA_100290+miR-29 inhibitor group”

The study uses human OSCC tissue and a xenograft model. Sex and demographics of the patients providing tissue are not reported. Age/weight/health status of patients is not reported, nor is the sex of the cell lines. For the nude mice, it states 'BALB/C-nu/nu, female and 5-week-old' but housing conditions are not described. Cell lines are identified by name and source but no authentication (e.g., STR profiling) or mycoplasma testing is mentioned. The species and strain of the oral keratinocytes are not reported.

Sub-criteria

demographicsNOT REPORTED

sex reportedNOT REPORTED

sex justifiedN/A

age weight healthNOT REPORTED

housing conditionsNOT REPORTED

species strain sourceINADEQUATE

Per reviewer (3) — the votes behind the verdict

WARNReviewer 1· 80% conf

Sex, age/weight/health of patients, and demographics are not reported. For the in vivo experiment, the sex and source of nude mice are reported, but strain and housing conditions are partially described. Cell lines are identified with source but not authenticated or tested for mycoplasma.

Evidence

absence[Methods, Tissue specimens]

Sex and age/weight/health of patients are not reported.

direct quote[Methods, Tissue specimens]

“Tumour formation experiment in nude mice (BALB/C-nu/nu, female and 5-week-old) was accomplished at experimental animal centre of Central South University.”

absence[Methods, Tissue specimens]

Housing conditions for nude mice are not reported.

WARNReviewer 2· 80% conf

Biological variables are partially reported; sex and age of mice are given, but human patient demographics are not reported, and housing conditions for mice are not described.

Evidence

direct quote[Materials and methods, Tissue specimens]

“Tumour formation experiment in nude mice (BALB/C-nu/nu, female and 5-week-old) was accomplished at experimental animal centre of Central South University.”

absence[Materials and methods, Tissue specimens]

Human patient demographics are not reported.

absence[Materials and methods, Tissue specimens]

Housing conditions for mice are not described.

WARNReviewer 3· 80% conf

Sex and age are reported for the animal model, but not for the human tissue samples. Strain/source is reported for mice and cell lines, but the human tissue lacks demographic details beyond being from patients at a single hospital.

Evidence

direct quote[Materials and methods, Tissue specimens]

“Tumour formation experiment in nude mice (BALB/C-nu/nu, female and 5-week-old)”

absence[Materials and methods, Tissue specimens]

Human tissue demographics are not reported.

direct quote[Materials and methods, Cell culture]

“The source of the three cell lines were derived from tongue cancer and have been authenticated in oral cancer research.”

The paper states 'The study was approved by the institutional review board of the Second Xiangya Hospital of Central South University for scientific and ethical integrity.' This is an adequate ethics approval statement. However, informed consent from patients is not mentioned. For the animal experiment, no IACUC approval or institutional animal welfare statement is provided; only the location of the experiment is given. Regulatory compliance is not addressed.

Sub-criteria

iacuc statementNOT REPORTED

informed consentNOT REPORTED

irb ethics statementADEQUATE

regulatory complianceNOT REPORTED

Per reviewer (3) — the votes behind the verdict

WARNReviewer 1· 90% conf

An IRB statement is provided for the human tissue study, but there is no mention of informed consent from patients. The animal experiment has no IACUC statement or regulatory compliance mention.

Evidence

direct quote[Methods, Tissue specimens]

“The study was approved by the institutional review board of the Second Xiangya Hospital of Central South University for scientific and ethical integrity.”

absence[Methods, all sections]

Informed consent is not mentioned.

absence[Methods, Tissue specimens]

No IACUC statement is provided for the animal experiment.

WARNReviewer 2· 90% conf

Ethical approval is mentioned for the human study, but the statement lacks a protocol number, and informed consent is not explicitly described. Animal ethics approval is not reported.

Evidence

direct quote[Materials and methods, Tissue specimens]

“The study was approved by the institutional review board of the Second Xiangya Hospital of Central South University for scientific and ethical integrity.”

absence[Materials and methods, Tissue specimens]

Informed consent is not reported.

absence[Materials and methods, Tissue specimens]

Animal ethics approval is not reported.

PASSReviewer 3· 90% conf

IRB approval for the human tissue study and IACUC approval for the animal study are stated, and no informed consent process is needed for de-identified archival tissue (though not explicitly stated, it's a minor omission).

Evidence

direct quote[Materials and methods, Tissue specimens]

“The study was approved by the institutional review board of the Second Xiangya Hospital of Central South University for scientific and ethical integrity.”

direct quote[Materials and methods, Tissue specimens]

“Tumour formation experiment in nude mice (BALB/C-nu/nu, female and 5-week-old) was accomplished at experimental animal centre of Central South University.”

absence[Materials and methods]

Informed consent for the human tissue samples is not reported.

The study uses multiple resources: cell lines (SCC9, CAL27, HN4, 293T, oral keratinocytes) are named with sources but no authentication (e.g., STR profiling) or mycoplasma testing is reported. Antibodies (anti-CDK6, anti-pRb, anti-GAPDH) are identified with catalog numbers and dilutions but no RRIDs. siRNAs and primers are identified with sequences. Luciferase and EGFP vectors are named but no catalog numbers. Microarray platform is described but not a RRID. Software (Cytoscape, Arraystar software) is named without versions.

Sub-criteria

mycoplasma testingNOT REPORTED

reagents identifiedINADEQUATE

organisms identifiedINADEQUATE

antibodies identifiedINADEQUATE

cell line authenticationNOT REPORTED

software tools identifiedINADEQUATE

Per reviewer (3) — the votes behind the verdict

WARNReviewer 1· 85% conf

Cell lines are identified by name and source but not authenticated or tested for mycoplasma. Antibodies are identified with catalog numbers and dilutions but not RRIDs. Key reagents (siRNAs, vectors, MTT) are identified but not fully cataloged. Software is mentioned but versions are lacking.

Evidence

direct quote[Methods, Cell culture]

“The human OSCC cell lines SCC9 and CAL27 were obtained from the Cancer Research Institute of Central South University (Changsha, China).”

direct quote[Methods, Immunohistochemistry and immunofluorescence analysis]

“primary antibodies (sc-7961, dilution 1:200, Santa Cruz Biotechnology...)”

WARNReviewer 2· 85% conf

Key resources are partially identified; antibodies and cell lines are named with sources, but validation data and catalog numbers are incomplete, and software versions are not reported.

Evidence

direct quote[Materials and methods, Immunohistochemistry and immunofluorescence analysis]

“the sections were incubated with primary antibodies (sc-7961, dilution 1:200, Santa Cruz Biotechnology, Santa Cruz, CA, USA)”

direct quote[Materials and methods, Cell culture]

“The human OSCC cell lines SCC9 and CAL27 were obtained from the Cancer Research Institute of Central South University (Changsha, China).”

direct quote[Materials and methods, ceRNA analysis and target prediction]

“We constructed a has_circRNA_100290-miRNA-target gene network using Cytoscape to visualize their interactions”

WARNReviewer 3· 85% conf

Antibodies, cell lines, reagents, and software are identified with catalog numbers and sources, but several details are missing: RRIDs, STR authentication for cell lines, full primer sequences for some miRNA primers, and all antibodies lack lot numbers.

Evidence

direct quote[Materials and methods, Immunohistochemistry]

“primary antibodies (sc-7961, dilution 1:200, Santa Cruz Biotechnology, Santa Cruz, CA, USA)”

direct quote[Materials and methods, Cell culture]

“The human OSCC cell lines SCC9 and CAL27 were obtained from the Cancer Research Institute of Central South University”

direct quote[Materials and methods, Microarray analysis]

“scanned by the Agilent Scanner G2505C”

The methods state 'One-way analysis of variance was applied to compare difference between two groups. A value of P <0.05 indicated that the difference was statistically significant.' This is a flaw (ANOVA for two groups is overkill). Assumptions (normality, equal variance) are not verified. Most p-values are reported as '<0.05' or '<0.01' rather than exact values. Effect sizes and confidence intervals are not reported. Data are presented as bar graphs with SD, but individual data points are not shown. Statistical software (Quantity One, ABI PRISM systems) is named without version. No mathematical plausibility issues are apparent.

Sub-criteria

tests namedINADEQUATE

exact p valuesNOT REPORTED

effect sizes ciNOT REPORTED

data presentationINADEQUATE

software identifiedINADEQUATE

assumptions verifiedNOT REPORTED

mathematical plausibilityN/A

Per reviewer (3) — the votes behind the verdict

WARNReviewer 1· 80% conf

Statistical tests are named (one-way ANOVA, t-test), and a threshold is set. However, test assumptions are not verified, exact p-values are largely not reported (only '<0.05' or '<0.01'), effect sizes and CIs are absent, and data presentation is partly adequate (means+SD shown) but individual data points are not shown. Software is identified (Quantity One, ABI PRISM) but not versioned.

Evidence

direct quote[Methods, Statistical analysis]

“One-way analysis of variance was applied to compare difference between two groups. A value of P <0.05 indicated that the difference was statistically significant.”

direct quote[Results, Figure legends and text]

“** P <0.01”

WARNReviewer 2· 80% conf

Statistical analysis is partially adequate; tests are named, but assumptions are not verified, exact p-values are not consistently reported, and effect sizes with confidence intervals are absent.

Evidence

direct quote[Materials and methods, Statistical analysis]

“One-way analysis of variance was applied to compare difference between two groups.”

direct quote[Results, circRNA_100290 and CDK6 are upregulated in OSCC]

“The presented values are the means±s.d., * P <0.05.”

absence[Results, circRNA_100290 and CDK6 are upregulated in OSCC]

Effect sizes and confidence intervals are not reported.

WARNReviewer 3· 75% conf

The statistical methods are described (one-way ANOVA), but key details are missing: exact p-values are rarely reported (mostly 'P<0.05'), effect sizes and confidence intervals are absent, individual data points are not shown (only means±SD), and normality/equal variance assumptions are not verified.

Evidence

direct quote[Figure legends and Results]

“* P <0.05 and ** P <0.01”

absence[Materials and methods, Statistical analysis]

Assumptions (normality, equal variance) not mentioned.

absence[Results and Figure legends]

Effect sizes or confidence intervals are not reported.

The paper does not include a data availability statement. The microarray data are not deposited in any public repository (e.g., GEO, ArrayExpress). No accession numbers are provided. Custom code (e.g., for ceRNA network analysis or statistical analysis) is not shared. The only URL given is the UCSC genome database, which is a reference, not study-specific data.

Sub-criteria

code sharingNOT REPORTED

accession numbersNOT REPORTED

repository depositNOT REPORTED

data availability statementNOT REPORTED

Per reviewer (3) — the votes behind the verdict

FAILReviewer 1· 95% conf

No data availability statement is provided, no microarray data deposited in a public repository, no accession numbers, and no code sharing.

Evidence

absence[All sections]

No data availability statement is present.

absence[All sections]

No repository deposit or accession numbers for microarray data.

absence[All sections]

No code sharing is mentioned.

FAILReviewer 2· 95% conf

No data or code availability is reported; the paper lacks a data availability statement, repository deposit, accession numbers, and code sharing.

Evidence

absence[Entire paper]

No data availability statement is present.

absence[Entire paper]

No repository deposit is mentioned.

absence[Entire paper]

No code sharing is mentioned.

FAILReviewer 3· 90% conf

No data or code availability statement is provided. Microarray data are not deposited in a public repository; no accession numbers or code are provided.

Evidence

absence[Entire paper]

No data availability statement.

absence[Entire paper]

No repository deposition of microarray data.

absence[Entire paper]

No code sharing or pipeline description.

Methods are detailed for cell culture, transfections, and assays, but lack critical design elements (see study_design). No reporting guideline (ARRIVE, CONSORT) is referenced. Not a clinical trial, so trial registration is n/a. All outcomes appear to be reported. Limitations are mentioned (off-target effects of siRNA, need for more specific methods). Conclusions are mainly about ceRNA mechanism and potential as a target, which is proportional to the evidence. Funding sources and grant numbers are provided, and a conflict of interest statement is present.

Sub-criteria

funding coiADEQUATE

trial registrationN/A

reporting guidelineNOT REPORTED

methods completenessINADEQUATE

all outcomes reportedADEQUATE

limitations discussedADEQUATE

conclusions proportionalADEQUATE

Per reviewer (3) — the votes behind the verdict

WARNReviewer 1· 85% conf

Methods are generally complete, but key details are missing (randomization, blinding, power analysis). No trial registration or reporting guideline is mentioned. Limitations are partially discussed, conclusions are largely proportional, and funding/COI statements are provided.

Evidence

direct quote[Discussion, paragraph 3]

“however, due to the difficulty of designing siRNA at the back-splice junction of circRNA, in this study, only one siRNA can specifically target the circular form of SLC30A7 but do not affect its linear form, thus off-target effects have to be considered.”

direct quote[Acknowledgments and disclosures]

“This study was supported by National Natural Science Foundation of China (Contract grant number: 81500832) and High Level Talent Program of Central South University (Contract grant number: 502035026).”

absence[Methods, all sections]

No reporting guideline is referenced.

WARNReviewer 2· 85% conf

Reporting is partially transparent; methods are detailed, and limitations are discussed, but the paper lacks a reporting guideline, trial registration, and a funding/COI statement is present but incomplete.

Evidence

direct quote[Materials and methods, Cell culture]

“DMEM/F12 medium supplemented with 0.4 μg/ml hydrocortisone was used to incubate the OSCC cell lines and α-MEM medium was used to culture the oral keratinocytes.”

direct quote[Discussion, paragraph 5]

“However, due to the difficulty of designing siRNA at the back-splice junction of circRNA, in this study, only one siRNA can specifically target the circular form of SLC30A7 but do not affect its linear form, thus off-target effects have to be considered.”

direct quote[End of paper]

“The authors declare no conflict of interest.”

WARNReviewer 3· 85% conf

Methods are generally detailed enough for replication, and conclusions are proportional, but there is no trial registration (not required), no reporting guideline referenced, and limitations are only briefly discussed at the end.

Evidence

absence[Materials and methods]

Reporting guidelines not mentioned.

direct quote[Discussion, last paragraph]

“only one siRNA can specifically target the circular form of SLC30A7 but do not affect its linear form, thus off-target effects have to be considered”

direct quote[End of paper]

18 copyedit issues flagged: mostly typo, consistency, punctuation.

Checked 61 references: 58 verified — 2 unresolved, 1 not checked.

Reproducibility

1 data/code link checked; 1 live.

Αlpha¹ Rigor Action ItemsFinal pass before journal submission

This published paper has significant reporting gaps that an informed reader should weigh: the missing data availability statement (microarray data not deposited) and absent animal ethics approval are the most consequential. The lack of randomization, blinding, and power analysis for the in vivo experiment limits the strength of causal claims. A correction or independent re-analysis would be warranted to address these issues.

1.
HIGHdata code
Deposit the microarray data in a public repository (e.g., GEO) and provide accession numbers in a dedicated Data Availability section.
The paper generates high-throughput data but provides no way for readers to access it, which is a fundamental reproducibility requirement.
2.
HIGHethics
Add an IACUC approval statement for the nude mouse experiment, including the institution and protocol number, in the Methods section (Tissue specimens).
Animal ethics approval is a mandatory reporting requirement for in vivo studies; its absence is a serious omission.
3.
HIGHethics
Describe the informed consent process for human tissue sample collection, including whether written consent was obtained, in the Methods section (Tissue specimens).
Informed consent is a standard ethical requirement for human tissue research; its absence is a reporting gap.
4.
HIGHreporting
Report exact p-values (e.g., P=0.023) instead of thresholds like P<0.05 throughout the Results section and figure legends.
Threshold-only p-values are insufficient for readers to assess the strength of evidence; exact values are standard.
5.
HIGHreporting
Provide a power analysis or sample size justification for the in vivo mouse experiment in the Methods section.
The xenograft experiment uses only 3 mice per group; a power analysis would justify this sample size or indicate a need for more.
6.
HIGHreporting
Describe the method of randomization for assigning mice to experimental groups and state whether blinding was used during tumor volume measurements or data analysis, in the Methods section (Tissue specimens).
Randomization and blinding are essential to minimize bias in animal experiments; their absence weakens causal inferences.
7.
HIGHreporting
Report effect sizes (e.g., Cohen's d) and confidence intervals for key comparisons in the Results section.
Effect sizes and CIs provide a more informative measure of the magnitude and precision of effects than p-values alone.
8.
HIGHreporting
Verify and report test assumptions (e.g., normality, equal variance) for ANOVA analyses in the Methods section (Statistical analysis).
Failure to verify assumptions can invalidate statistical conclusions; this is a standard reporting requirement.
9.
HIGHreporting
Include a statement on mycoplasma testing for the cell lines used in the Methods section (Cell culture).
Mycoplasma contamination can affect experimental results; its absence is a common reviewer concern.
10.
HIGHreporting
Provide catalog numbers and RRIDs for all antibodies and key reagents in the Methods section.
RRIDs enable unambiguous identification of reagents and improve reproducibility.
11.
HIGHreporting
Report the version numbers of all software used (e.g., Cytoscape 3.8.0, Arraystar version) in the Methods section.
Software versions are necessary for reproducibility of bioinformatic analyses.
12.
HIGHreporting
Define the distinction between biological and technical replicates for qPCR, Western blots, and cell proliferation assays in the Methods section (Statistical analysis).
Clear replicate definitions are essential for assessing the reliability of the data.
13.
HIGHreporting
Report the sex and age/weight/health status of the patients who provided tissue samples in the Methods section (Tissue specimens).
Patient demographics are important for assessing generalizability and potential confounding.
14.
HIGHreporting
Correct the statistical method description: 'One-way analysis of variance was applied to compare difference between two groups' – if only two groups are compared, a t-test is more appropriate; if multiple groups, specify which. Revise in Methods (Statistical analysis).
The current description is methodologically inconsistent and could confuse readers.
15.
MEDIUMcopyedit
Fix the title: remove the leading '1' and ensure it is a single coherent sentence.
The title has a numbering artifact that appears unprofessional.
16.
MEDIUMcopyedit
Format the author list and affiliations properly with commas and line breaks in the abstract.
The current formatting is run together and difficult to read.
17.
MEDIUMcopyedit
Standardize the spelling of the cell line 'HN4' (not 'NH4') throughout the manuscript.
Inconsistent spelling of a key resource is confusing.
18.
MEDIUMcopyedit
Remove the duplicate URL in the Results section: 'http://genome.ucsc.edu/ http://genome.ucsc.edu/'.
Duplicate URL is a typographical error.
19.
MEDIUMcopyedit
Change 'Immunochemistry' to 'Immunohistochemistry' in the Results section for consistency with the Methods.
Inconsistent terminology can confuse readers.
20.
MEDIUMcopyedit
Change 'an secondary antibody' to 'a secondary antibody' in the Methods section (Immunohistochemistry).
Grammatical error.
21.
LOWcopyedit
Ensure figure references are consistently formatted (e.g., 'Figure 3d') throughout the Results section.
Missing or inconsistent figure references reduce clarity.
22.
LOWcopyedit
Use italic P and consistent spacing (e.g., P < 0.05) throughout the manuscript.
Inconsistent formatting of p-values is a minor but noticeable issue.

Full text (52,517 chars)Analyzed: Jul 6, 2026

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