Verified Rigor RecordRigor Reviewed

Rigor-Reviewed Manuscript

JQ1 suppresses tumor growth via PTEN/PI3K/AKT pathway in endometrial cancer · 10.18632/oncotarget.11631

Name: Verified Rigor Record — JQ1 suppresses tumor growth via PTEN/PI3K/AKT pathway in endometrial cancer · 10.18632/oncotarget.11631
Item: JQ1 suppresses tumor growth via PTEN/PI3K/AKT pathway in endometrial cancer · 10.18632/oncotarget.11631
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 · Interventional Study

Eight dimensions — click a segment for details

Critical

High-priority

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

Not verified in this report

References were not verified against Crossref/OpenAlex.
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.

This preclinical study evaluates JQ1 as a c-Myc inhibitor in endometrial cancer, with a well-grounded scientific premise. However, the methods reporting is critically incomplete: key design elements (randomization, blinding, power analysis) are absent, statistical reporting is minimal (no effect sizes, no assumption checks), and resource identification (antibodies, cell line authentication) is incomplete. The paper would benefit from substantial methodological reporting improvements before the findings can be considered fully robust.

Evaluated based on three independent reviewer assessments, a copyedit pass, and verification components (statistics, reproducibility, preregistration, integrity, claim audit). The statistics component found no recomputable tests; the integrity check flagged an internal contradiction between mouse strains used and an implausible dose selection. Preregistration was not applicable.

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

2 integrity concerns flagged (0 high).

Rigor

Eight Dimensions1 pass · 6 warn · 1 fail

12 major claims1 partial11 supported

The introduction extensively cites prior work on c-Myc overexpression in endometrial cancer (30–50% of patients, references 14–15), JQ1 as a BRD4/c-Myc inhibitor (reference 21), and existing evidence of JQ1 efficacy in other cancers (references 22–24). The limitations of prior targeting strategies (technical difficulties, drug delivery, redundancy) are acknowledged (paragraph 4). The hypothesis that PTEN status may modulate JQ1 sensitivity follows logically from the observation that PTEN loss is the most common alteration in type I endometrial cancer (Results, paragraph beginning 'Given that PTEN loss'). All three sub-criteria are adequately addressed.

Sub-criteria

prior work citedADEQUATE

premise rationaleADEQUATE

limitations addressedADEQUATE

Per reviewer (3) — the votes behind the verdict

PASSReviewer 1· 95% conf

The premise that c-Myc is a promising target in endometrial cancer and that JQ1 inhibits c-Myc is well supported by prior literature, and the study rationale linking PTEN status to JQ1 sensitivity is clearly developed.

Evidence

direct quote[Introduction, paragraph 2]

“Overexpression of c-Myc is observed in 30–50% of patients with endometrial cancer and is associated with advanced stage, higher grade, distant metastasis and worse prognosis [, ].”

direct quote[Introduction, paragraph 4]

“Unfortunately, most of these approaches continue to be hampered by technical difficulties, pertaining largely to drug delivery and the fact that many c-Myc target genes are functionally redundant and/or cell type specific [].”

paraphrase[Results, paragraph beginning 'Given that PTEN loss']

“The authors note that PTEN loss is the most common genetic abnormality in type I endometrial cancer, motivating the investigation of PTEN status as a determinant of JQ1 sensitivity.”

PASSReviewer 2· 90% conf

The paper establishes a clear scientific foundation: it cites prior work on c-Myc overexpression in endometrial cancer, the development of JQ1 as a BRD4 inhibitor, and articulates a logical hypothesis linking JQ1 to PTEN status.

Evidence

direct quote[Introduction, paragraphs 2-3]

“Overexpression of c-Myc is observed in 30–50% of patients with endometrial cancer and is associated with advanced stage, higher grade, distant metastasis and worse prognosis”

direct quote[Introduction, paragraph 4]

“a small molecule, JQ1, was shown to be a potent c-Myc inhibitor. JQ1 was preliminarily designed as an inhibitor of bromodomain-containing proteins (BRDs)”

direct quote[Introduction, paragraph 4]

PASSReviewer 3· 95% conf

The paper clearly establishes the scientific foundation by citing prior work on c-Myc, JQ1, and PTEN, and provides a logical rationale for the study.

Evidence

direct quote[Introduction, paragraph 1-4]

“Overexpression of c-Myc is observed in 30–50% of patients with endometrial cancer and is associated with advanced stage, higher grade, distant metastasis and worse prognosis.”

direct quote[Introduction, paragraph 4]

direct quote[Introduction, paragraph 5]

“In the present study, we investigated the effects of JQ1 on cell proliferation and tumor growth in endometrial cancer.”

The in vitro experiments use a standard panel of cell lines (ATCC), with appropriate controls (e.g., DMSO, empty plasmids). The in vivo xenograft experiments use nude mice (n=4 per group) and describe JQ1 dosing (50 mg/kg/day) but do not report any randomization method for allocating mice to treatment groups or any blinding during tumor measurement. No power analysis or sample size justification is provided. Inclusion/exclusion criteria for the primary culture samples are not pre-specified: the paper states '28 consecutive pathologically diagnosed endometrial cancer samples were collected' but no other eligibility criteria. Outlier handling is not discussed. Biological vs. technical replicates are not explicitly distinguished for the in vitro experiments, though 'triplicate' is mentioned for MTT and cell cycle assays. Independent replication is absent (the xenograft experiments are not replicated).

Sub-criteria

controlsADEQUATE

power analysisNOT REPORTED

blinding levelsNOT REPORTED

outlier handlingNOT REPORTED

randomization unitNOT REPORTED

inclusion exclusionINADEQUATE

randomization methodNOT REPORTED

replicate distinctionINADEQUATE

independent replicationNOT REPORTED

Per reviewer (3) — the votes behind the verdict

WARNReviewer 1· 90% conf

The study design, combining in vitro and in vivo experiments, is well-structured but has critical gaps: no randomization or blinding is reported, no power analysis, and no pre-specified inclusion/exclusion criteria for the xenograft experiments or primary cultures.

Evidence

absence[Materials and Methods, Tumor xenografts growth assay]

No randomization method is described for the xenograft experiments.

absence[Materials and Methods, Tumor xenografts growth assay]

No blinding of investigators during tumor measurement is reported.

direct quote[Materials and Methods, Tissue samples collection and primary cell culture]

“Twenty-eight consecutive pathologically diagnosed endometrial cancer samples were collected from UNC Hospital...”

WARNReviewer 2· 80% conf

The study design has multiple gaps: no randomization method for cell culture or animal experiments, no blinding described, no a priori power analysis, and incomplete reporting on outlier handling and replication.

Evidence

absence[Methods: Tumor xenografts growth assay]

No randomization method for animal experiments is described.

absence[Methods: all sections]

Blinding is not mentioned for any experiment.

direct quote[Methods: Tissue samples collection and primary cell culture]

“Twenty-eight consecutive pathologically diagnosed endometrial cancer samples were collected from UNC Hospital ... and the First Affiliated Hospital of Zhengzhou University”

FAILReviewer 3· 95% conf

The study lacks essential design elements such as randomization, blinding, power analysis, and explicit handling of replicates and outliers, resulting in a failing score.

Evidence

absence[Materials and Methods, Tumor xenografts growth assay]

No randomization method is described for animal experiments.

absence[Materials and Methods]

Blinding is not mentioned for any experiment.

absence[Materials and Methods]

No power analysis or sample size justification is provided.

For the in vivo xenograft model, mice are described as '6-week-old female mice' (sex and approximate age reported), but weight, health status, and housing conditions (light cycle, temperature, enrichment) are not reported. The species is specified as 'severe combined immunodeficiency' mice, but strain/substrain and source are not given beyond 'nude mice' and 'Balb b/c nude mice' (two different names are used inconsistently). For the primary culture patients, the paper provides stage, grade, and histologic subtype for each case (Table 1), but does not report patient demographics (age, race/ethnicity, comorbidities) except indirectly through disease characteristics. The study includes both sexes (all female mice; human endometrial cancer patients are female), so sex_justified is not applicable.

Sub-criteria

demographicsNOT REPORTED

sex reportedADEQUATE

sex justifiedN/A

age weight healthINADEQUATE

housing conditionsNOT REPORTED

species strain sourceINADEQUATE

Per reviewer (3) — the votes behind the verdict

WARNReviewer 1· 85% conf

The paper reports sex, age (for human subjects), and some health status for mice, but lacks key details: animal housing conditions, specific ages/weights of mice, and detailed demographics for the primary culture patients.

Evidence

direct quote[Materials and Methods, Tumor xenografts growth assay]

“Six-week-old female mice with severe combined immunodeficiency were used for this assay.”

absence[Materials and Methods]

Housing conditions (light cycle, temperature, enrichment) are not reported.

absence[Results, Table 1]

Patient demographics (age, race, comorbidities) are not reported in Table 1 or elsewhere.

WARNReviewer 2· 85% conf

Sex and species/strain are reported (female mice, six cell lines from ATCC), but age/weight/health status of mice, housing conditions, and human demographics are incomplete.

Evidence

direct quote[Methods: Tumor xenografts growth assay]

“Six-week-old female mice with severe combined immunodeficiency were used for this assay.”

absence[Methods: Tumor xenografts growth assay]

Housing conditions not described.

paraphrase[Results: Table 1]

“Table 1 lists stage, grade, histologic subtype, IC50, and PTEN status for 28 patients; no race/ethnicity or comorbidities.”

WARNReviewer 3· 85% conf

Sex and age are reported, but species/strain source is incomplete, housing conditions are missing, and single-sex use is not justified.

Evidence

direct quote[Materials and Methods, Tumor xenografts growth assay]

“Six-week-old female mice with severe combined immunodeficiency were used for this assay.”

direct quote[Results, The anti-tumor activity of JQ1 depended on PTEN status in vivo]

“we established four nude mice models (Balb b/c nude mice)”

absence[Materials and Methods]

Housing conditions (e.g., light cycle, enrichment) are not described.

For human tissue samples, the paper states 'All patients provided written consents and approval was obtained from the respective Institutional Review Board (IRB) committees' — this names the type of body but not the specific committee(s), institution(s), or protocol numbers, making it reported_but_inadequate. For the xenograft experiments, the paper states 'All mice were handled according to protocols approved by the ethics committee of Zhengzhou University' — again, no protocol number. Regulatory compliance: the paper does not explicitly state adherence to the Declaration of Helsinki, the Guide for the Care and Use of Laboratory Animals, or any other named framework, so this is not_reported.

Sub-criteria

iacuc statementINADEQUATE

informed consentADEQUATE

irb ethics statementINADEQUATE

regulatory complianceNOT REPORTED

Per reviewer (3) — the votes behind the verdict

WARNReviewer 1· 90% conf

Ethics approval for the primary culture samples is reported but the statement is vague: a named approving body is not given for the IRB, and the animal statement lacks a protocol number. For the xenograft experiments, the ethics committee is named but no approval number is provided.

Evidence

direct quote[Materials and Methods, Tissue samples collection and primary cell culture]

“All patients provided written consents and approval was obtained from the respective Institutional Review Board (IRB) committees.”

direct quote[Materials and Methods, Tumor xenografts growth assay]

“All mice were handled according to protocols approved by the ethics committee of Zhengzhou University.”

absence[Materials and Methods]

No statement of regulatory compliance (e.g., Declaration of Helsinki, Guide for Care and Use of Laboratory Animals) is found.

PASSReviewer 2· 90% conf

Ethical approvals are adequately reported: IRB approval for human primary cultures, IACUC for animal work, and patient consent are described.

Evidence

direct quote[Methods: Tissue samples collection and primary cell culture]

“All patients provided written consents and approval was obtained from the respective Institutional Review Board (IRB) committees.”

direct quote[Methods: Tumor xenografts growth assay]

“All mice were handled according to protocols approved by the ethics committee of Zhengzhou University.”

WARNReviewer 3· 90% conf

Ethics approvals for human and animal studies are stated, but regulatory compliance with a named framework (e.g., Declaration of Helsinki) is not explicitly mentioned.

Evidence

direct quote[Materials and Methods, Tissue samples collection and primary cell culture]

“All patients provided written consents and approval was obtained from the respective Institutional Review Board (IRB) committees.”

direct quote[Materials and Methods, Tumor xenografts growth assay]

“All mice were handled according to protocols approved by the ethics committee of Zhengzhou University.”

absence[Materials and Methods]

No explicit mention of the Declaration of Helsinki or other regulatory framework.

All six cell lines are sourced from ATCC (vendor named) but specific catalogue numbers or RRIDs are not provided, and no authentication (e.g., STR profiling) is reported. No mycoplasma testing is mentioned. Antibodies are identified by target and vendor (Cell Signaling Technology, Sigma-Aldrich), with dilutions stated (e.g., 1:1000 for primary antibodies), but no catalogue numbers, clone IDs, or RRIDs are given — this is reported_but_inadequate. Reagents: JQ1 is from Dr. James E. Brander (no catalogue number); pcDNA3-PTEN and shRNA-PTEN plasmids from Addgene with catalogue numbers (10759 and 10669). Software: FCS Express 4 (De Novo Software, version stated), Cellometer (Nexcelom), and GenePix Pro 4.1 are identified with vendors, but versions are missing for some. Microarray platform: UNC-customized microarray from Agilent, but no accession number or RRID. The investigational product (JQ1) is identified by name and source, which is adequate for a small-molecule drug (the paper is a preclinical study of a drug, not a clinical trial, so drug identification is applicable).

Sub-criteria

mycoplasma testingNOT REPORTED

reagents identifiedADEQUATE

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 and reagents are adequately identified, but critical gaps include no mycoplasma testing, no STR authentication for cell lines, and incomplete identification of antibodies (dilutions given but no catalogue or lot numbers).

Evidence

direct quote[Materials and Methods, Cancer cell lines and reagents]

“All the primary antibodies for phosphorylated-S6, pan-S6, phosphorylated-AKT, pan-AKT, cyclin D1, CDK6, CDK4 and Ki-67 were obtained from Cell Signaling Technology (Danvers, MA, USA).”

absence[Materials and Methods, Cancer cell lines and reagents]

No mycoplasma testing is reported for any of the cell lines used.

direct quote[Materials and Methods, Establishment of the stably transfected cell lines]

“The pcDNA3-PTEN and shRNA-PTEN plasmids were obtained from Addgene (plasmid 10759 and 10669, Cambridge, MA).”

WARNReviewer 2· 85% conf

Critical resources are partially identified: antibodies have vendor but no catalog numbers; JQ1 source is stated vaguely; cell lines are from ATCC; plasmids from Addgene are identified; software lacks version numbers.

Evidence

direct quote[Methods: Cancer cell lines and reagents]

“All the primary antibodies for phosphorylated-S6, pan-S6, phosphorylated-AKT, pan-AKT, cyclin D1, CDK6, CDK4 and Ki-67 were obtained from Cell Signaling Technology (Danvers, MA, USA).”

direct quote[Methods: Cancer cell lines and reagents]

“All cells were purchased from ATCC (American type culture collection, USA)”

absence[Methods: all sections]

Mycoplasma testing not mentioned.

FAILReviewer 3· 95% conf

Critical resources such as antibodies, cell lines, and reagents are inadequately identified, with no catalog numbers, authentication, or mycoplasma testing reported.

Evidence

direct quote[Materials and Methods, Cancer cell lines and reagents]

“All the primary antibodies for phosphorylated-S6, pan-S6, phosphorylated-AKT, pan-AKT, cyclin D1, CDK6, CDK4 and Ki-67 were obtained from Cell Signaling Technology (Danvers, MA, USA).”

direct quote[Materials and Methods, Cancer cell lines and reagents]

“All cells were purchased from ATCC (American type culture collection, USA)”

absence[Materials and Methods]

Mycoplasma testing is not reported.

The 'Statistical analysis' section states: 'Results were compared by Student's t test and data were expressed as mean ± S.E. χ² test was used for the analysis of 2×2 tables.' This names the tests, but no verification of assumptions (normality, equal variance) is reported. Some exact p-values are given (e.g., p=0.009, p=0.021), but many are reported as inequalities (p<0.001, p<0.05). No effect sizes (e.g., Cohen's d, hazard ratios) or confidence intervals are reported for any comparison. Software is identified (FCS Express 4, Cellometer) but the statistical analysis section does not name a specific package (e.g., SPSS, GraphPad Prism) for the t-tests and χ² tests. Data presentation: bar charts with error bars are used, but error bars are not defined (SEM/CI), and individual data points are not shown. Per-group n is stated for some comparisons (e.g., n=4 mice per group) but inconsistently. Mathematical plausibility checks: the IC50 values and percentages from Table 1 are plausible. No arithmetic errors are detected. Overall, the reporting is suboptimal but not demonstrably erroneous.

Sub-criteria

tests namedINADEQUATE

exact p valuesINADEQUATE

effect sizes ciNOT REPORTED

data presentationINADEQUATE

software identifiedINADEQUATE

assumptions verifiedNOT REPORTED

mathematical plausibilityPASS

Per reviewer (3) — the votes behind the verdict

WARNReviewer 1· 80% conf

Statistical tests are named (Student's t-test, χ²), but key details are missing: no verification of test assumptions, no effect sizes or confidence intervals, and exact p-values are not consistently reported (many are given as '<0.05' rather than precise values). Software is identified but version is minimal.

Evidence

direct quote[Materials and Methods, Statistical analysis]

“Results were compared by Student's t test and data were expressed as mean ± S.E. χ2 test was used for the analysis of 2×2 tables.”

direct quote[Results, multiple sections]

“JQ1 significantly suppressed tumor growth and reduced tumor weight in Hec-1a xenografts compared to control mice after 4 weeks of treatment (p < 0.001).”

absence[Results]

No confidence intervals or effect sizes are reported.

WARNReviewer 2· 75% conf

Statistical tests are named (Student's t, χ²), but exact p-values are often not reported (ranges given), effect sizes/CI missing, assumptions not verified, and individual data points are not shown.

Evidence

direct quote[Methods: Statistical analysis]

“Results were compared by Student's t test and data were expressed as mean ± S.E. χ2 test was used for the analysis of 2 × 2 tables.”

direct quote[Results, multiple figures]

“JQ1 notably suppressed tumor growth and reduced tumor weight ... (p < 0.001)”

absence[Results and Methods]

Effect sizes and confidence intervals not reported.

FAILReviewer 3· 90% conf

While statistical tests are named, the paper lacks essential reporting: effect sizes, confidence intervals, software identification, and verification of assumptions. P-values are often given as inequalities.

Evidence

direct quote[Materials and Methods, Statistical analysis]

“Results were compared by Student's t test and data were expressed as mean ± S.E. χ2 test was used for the analysis of 2 × 2 tables.”

direct quote[Results, JQ1 inhibited cell proliferation in Hec-1a and KLE cells]

“JQ1 (100 nM) notably suppressed the colon formation in Hec-1a and KLE cells (p = 0.009 and 0.021 respectively).”

absence[Results and Tables]

Effect sizes and confidence intervals are not reported.

The paper mentions that microarray data are uploaded to a UNC database (https://genome.unc.edu/), but no accession number or DOI is provided. No data availability statement covers other data (cell line, animal, primary culture). No code is shared. All four sub-criteria are inadequate, leading to a fail.

Sub-criteria

code sharingNOT REPORTED

accession numbersNOT REPORTED

repository depositINADEQUATE

data availability statementINADEQUATE

Per reviewer (3) — the votes behind the verdict

WARNReviewer 1· 85% conf

A data availability statement is absent. Microarray data were uploaded to a UNC database, but no persistent identifier or public accession number is provided. No statement about patient-level data is made. Custom code was not used, so code_sharing is not applicable.

Evidence

absence[Entire manuscript]

No data availability statement is present anywhere in the manuscript.

direct quote[Materials and Methods, Microarray analysis]

“The results were analyzed with GenePix Pro 4.1 and uploaded onto the Microarray Database of UNC at Chapel Hill (https://genome.unc.edu/).”

PASSReviewer 2· 85% conf

Microarray data are deposited and accessible via a link; no data availability statement for other data, but code sharing is not applicable.

Evidence

direct quote[Methods: Microarray analysis]

“The results were analyzed with GenePix Pro 4.1 and uploaded onto the Microarray Database of UNC at Chapel Hill (https://genome.unc.edu/).”

FAILReviewer 3· 95% conf

Data availability is limited to a vague statement about microarray data, with no persistent identifiers, and no code sharing.

Evidence

direct quote[Materials and Methods, Microarray analysis]

“The results were analyzed with GenePix Pro 4.1 and uploaded onto the Microarray Database of UNC at Chapel Hill (https://genome.unc.edu/).”

absence[Entire paper]

No data availability statement for other data types.

absence[Entire paper]

No code sharing is mentioned.

Methods are detailed and reproducible (reagent details, cell culture conditions, antibody dilutions, assay protocols). The paper does not reference any reporting guideline (ARRIVE for animal studies, CONSORT for clinical trials, etc.). Limitations are discussed in the Discussion (e.g., PTEN status as predictive marker may be cancer-type specific, need for combination therapy), but the discussion does not address important methodological limitations such as the lack of blinding, small sample size (n=4 mice per group), or the use of a single xenograft model. Conclusions are proportional: the paper states that JQ1 'might serve as a novel therapeutic strategy in PTEN-positive endometrial cancers,' which is appropriately cautious. The funding sources (Steelman Fund, NIH, National Natural Science Funds of China) and conflicts of interest (none reported) are stated. All outcomes mentioned in the Results appear to be reported, including negative/null results (e.g., PTEN-negative cell lines were resistant).

Sub-criteria

funding coiADEQUATE

trial registrationN/A

reporting guidelineNOT REPORTED

methods completenessADEQUATE

all outcomes reportedADEQUATE

limitations discussedINADEQUATE

conclusions proportionalADEQUATE

Per reviewer (3) — the votes behind the verdict

WARNReviewer 1· 90% conf

Methods are described in sufficient detail for replication, but critical pieces are missing: no trial registration (not required for preclinical study), no reporting guideline mentioned, and the limitations discussion is present but does not address key sources of bias (e.g., lack of blinding). Conclusions are proportional to the evidence. Funding and COI are stated.

Evidence

absence[Materials and Methods]

No reporting guideline (ARRIVE, STROBE, etc.) is referenced.

paraphrase[Discussion]

“The Discussion focuses on biological limitations (PTEN status as a predictive marker depending on cancer type) but does not discuss methodological limitations such as the absence of blinding or the small group sizes.”

direct quote[Acknowledgements and Funding, and Footnotes]

“This work was generously supported by the Steelman Fund ... CONFLICTS OF INTEREST The authors have no potential conflicts of interest to report.”

WARNReviewer 2· 85% conf

Methods are detailed enough for replication, funding and COI are stated, and limitations are discussed, but no reporting guideline is referenced, trial registration is not applicable, and conclusions are proportional, but pre-specified outcomes are not explicitly listed.

Evidence

paraphrase[Methods: all sections]

“Methods include detailed protocols for cell culture, MTT assay, colony formation, cell cycle, apoptosis, Western blot, IHC, microarray, and xenografts.”

direct quote[Acknowledgments and Funding]

“This work was generously supported by the Steelman Fund (Bae-Jump VL). ... The authors have no potential conflicts of interest to report.”

absence[Methods: all sections]

No reporting guideline referenced.

PASSReviewer 3· 85% conf

The paper provides detailed methods, reports all outcomes, and has proportional conclusions with funding disclosure, but lacks a reporting guideline and has a limited limitations discussion.

Evidence

direct quote[Materials and Methods]

“Total protein was extracted from tumor cells or tissues using RIPA buffer (Thermo Fisher, MA) and quantified by BCA assay (Thermo Fisher).”

direct quote[Discussion]

“Further analysis found that c-Myc protein expression was not associated with sensitivity to JQ1 in these cells (data not shown).”

direct quote[Acknowledgments and Funding, Footnotes]

“This work was generously supported by the Steelman Fund (Bae-Jump VL).”

9 copyedit issues flagged: mostly consistency, typo, grammar.

Reproducibility

1 data/code link checked; 0 live.

Αlpha¹ Rigor Action ItemsFinal pass before journal submission

This post-publication audit identifies serious methodological and reporting deficiencies that undermine the robustness of the published findings. An informed reader should weigh the lack of randomization, blinding, and power analysis in the xenograft experiments, the incomplete statistical reporting, and the internal contradictions (mouse strain, IC50 vs. dose). These issues would warrant consideration of an erratum or independent re-analysis.

1.
HIGHrigor
Add randomization method and unit (e.g., computer-generated random numbers for mouse allocation) to the 'Tumor xenografts growth assay' section.
Without randomization, the animal experiments are at high risk of selection bias, invalidating statistical inferences.
2.
HIGHrigor
Describe blinding procedures for treatment administration and tumor measurements in the 'Tumor xenografts growth assay' section; if not feasible, state this and provide a rationale.
Lack of blinding is a major source of bias in preclinical studies, especially for subjective outcomes like tumor volume.
3.
HIGHstatistics
Report effect sizes (e.g., Cohen's d) and 95% confidence intervals for all primary and secondary comparisons in the Results section and figure legends.
P-values alone do not indicate the magnitude or precision of effects, making the results non-reproducible and incomplete.
4.
HIGHstatistics
Provide a complete data availability statement covering all data types (cell line, animal, primary culture) in a dedicated 'Data Availability' section, and deposit microarray data with a persistent identifier (e.g., GEO accession).
Without a full data availability statement, the data are not findable or reusable, which is a failure of modern research reporting.
5.
HIGHrigor
Resolve the internal contradiction: state which mouse strain was actually used ('severe combined immunodeficiency' or 'Balb/c nude') in the 'Tumor xenografts growth assay' and ensure consistency throughout.
The discrepancy between Methods and Results undermines confidence in the animal model used and the reproducibility of the experiments.
6.
HIGHrigor
Provide a power analysis or sample size justification for the xenograft experiments, specifying the expected effect size, alpha, and power, in the 'Tumor xenografts growth assay' section.
With only 4 mice per group, the study is likely underpowered to detect meaningful effects, and a justification is essential for interpretation.
7.
HIGHstatistics
Replace threshold p-values (e.g., p < 0.001) with exact p-values in the Results section and figures.
Exact p-values allow readers to assess the evidence quantitatively; reporting only thresholds is considered inadequate in modern science.
8.
HIGHrigor
Add catalog numbers, clone IDs, or RRIDs for all antibodies in the 'Cancer cell lines and reagents' section.
Without these identifiers, other researchers cannot obtain the same reagents, compromising reproducibility.
9.
HIGHrigor
Report mycoplasma testing and STR authentication for all cell lines in the 'Cancer cell lines and reagents' section, or state the reason if not performed.
Mycoplasma contamination and cell line misidentification are widespread and can invalidate experimental results.
10.
HIGHreporting
Cite the ARRIVE guidelines for animal experiments and reference them in the Methods section.
Following reporting guidelines improves completeness and transparency, and many journals now require it.
11.
MEDIUMreporting
Expand the limitations discussion in the Discussion section to address the lack of blinding, randomization, small sample size, and the use of only one xenograft model.
Acknowledging these methodological limitations is important for a balanced interpretation of the study.
12.
MEDIUMcopyedit
Correct the typo in the Abstract: change 'Q1' to 'JQ1'.
A typo in the title/abstract is unprofessional and may lead to indexing errors.
13.
MEDIUMcopyedit
Add the missing reference number in the Results paragraph beginning 'We previously reported' (empty bracket).
Incomplete citations leave readers unable to verify the claim.
14.
LOWcopyedit
Fix the typo 'cell s' to 'cells' in the Materials and Methods, Cancer cell lines and reagents section.
Minor typographical errors detract from the paper's professionalism.

Full text (52,467 chars)Analyzed: Jul 6, 2026

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