Verified Rigor RecordRigor Reviewed

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Loss of FOXN3 in colon cancer activates beta-catenin/TCF signaling and promotes the growth and migration of cancer cells · 10.18632/oncotarget.14189

Name: Verified Rigor Record — Loss of FOXN3 in colon cancer activates beta-catenin/TCF signaling and promotes the growth and migration of cancer cells · 10.18632/oncotarget.14189
Item: Loss of FOXN3 in colon cancer activates beta-catenin/TCF signaling and promotes the growth and migration of cancer cells · 10.18632/oncotarget.14189
Rating: 50
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: lowEngine: 7.6.0Source: full text

Not verified in this report

No reported statistical tests were found to recompute.
No data or code availability links were detected to verify.

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.

9 major claims checked against the paper's own evidence: 1 not fully backed by the presented evidence (unsupported or overstated).

2 integrity concerns flagged (0 high).

Rigor

Eight Dimensions1 pass · 2 warn · 5 fail

9 major claims1 overstated8 supported

The introduction cites prior research on FOXN3 down-regulation in various cancers and beta-catenin signaling in colon cancer, establishing a logical premise. The study addresses some gaps in understanding FOXN3's molecular mechanisms but does not discuss limitations of prior work in detail. The rationale linking FOXN3 down-regulation to beta-catenin/TCF signaling is clearly stated.

Sub-criteria

prior work citedADEQUATE

premise rationaleADEQUATE

limitations addressedINADEQUATE

Per reviewer (2) — the votes behind the verdict

PASSReviewer 1· 90% conf

The paper adequately cites prior work on FOXN3 and beta-catenin signaling, establishes a clear rationale for studying FOXN3 in colon cancer, and partially addresses limitations of prior research.

Evidence

direct quote[Introduction, paragraph 3]

“Several studies have reported the physiological functions of FOXN3 in the embryonic development [12,13]. Recently, down-regulation of FOXN3 has been observed in several cancer types, including hepatocellular carcinoma, oral squamous cell carcinoma, laryngeal cancer and diffuse large B-cell lymphoma [14–17].”

direct quote[Discussion]

“In this study, we explored the expression pattern of FOXN3 in colon cancer.”

direct quote[Introduction, paragraph 3]

“However, the molecular mechanisms through which down-regulation of FOXN3 promoted the tumorigenesis remain poorly understood.”

PASSReviewer 2· 90% conf

The paper adequately establishes the scientific premise by citing prior work on FOXN3 and beta-catenin signaling and providing a logical rationale.

Evidence

direct quote[Introduction, paragraph 2]

“Recently, down-regulation of FOXN3 has been observed in several cancer types, including hepatocellular carcinoma, oral squamous cell carcinoma, laryngeal cancer and diffuse large B-cell lymphoma [–]. However, the molecular mechanisms through which down-regulation of FOXN3 promoted the tumorigenesis remain poorly understood.”

direct quote[Abstract and Introduction]

“Aberrant activation of beta-catenin/TCF is a hallmark of colon cancer. How the functions of nuclear localized beta-catenin are regulated is not fully understood.”

absence[Introduction]

Limitations of prior work are not explicitly addressed.

Only controls are adequately described. Replicate distinction is partially addressed but not clearly defined as biological vs technical. The absence of randomization, blinding, power analysis, and outlier handling reduces confidence in the results.

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 (2) — the votes behind the verdict

WARNReviewer 1· 85% conf

The study design lacks randomization, blinding, and power analysis criteria applicable to cell-line experiments; inclusion/exclusion criteria and outlier handling are not reported for the human tissue samples.

Evidence

direct quote[Materials and Methods, MTT assay]

“All experiments were performed in triplicates.”

paraphrase[Materials and Methods, Statistical analysis]

“Statistical methods are described but no power analysis is mentioned.”

paraphrase[Materials and Methods, Clinical colon cancer tissues]

“The method states tissues were collected from patients who received surgery but no specific criteria are given.”

FAILReviewer 2· 95% conf

The study design lacks many standard rigor elements such as randomization, blinding, power analysis, and explicit inclusion/exclusion criteria.

Evidence

paraphrase[Results, Figures 2-4]

“Empty vector or siControl were used as negative controls; ICG001 was used as an inhibitor of beta-catenin/TCF signaling.”

direct quote[Materials and Methods, MTT assay]

“All experiments were performed in triplicates.”

absence[Materials and Methods]

No randomization, blinding, power analysis, or inclusion/exclusion criteria are reported.

The mouse model is described genetically but no details on sex, age, housing, or health status are provided. Cell lines are sourced from ATCC, which is adequate for species/strain, but other biological variables are missing.

Sub-criteria

demographicsN/A

sex reportedNOT REPORTED

sex justifiedNOT REPORTED

age weight healthNOT REPORTED

housing conditionsNOT REPORTED

species strain sourceADEQUATE

Per reviewer (2) — the votes behind the verdict

WARNReviewer 1· 80% conf

Sex and age/weight/health of patients are not reported; the mouse model is not described in sufficient detail (sex, age, weight, housing conditions not provided).

Evidence

direct quote[Materials and Methods, Cell culture]

“Colon cancer cell lines (SW480, SW620 and HCT8) and normal human colon epithelial cell line (HIEC and NCM460) were obtained from ATCC (American Typical Culture Center).”

paraphrase[Materials and Methods, Mice model]

“The mice model section describes the genetic model but does not mention sex, age, or weight of mice.”

paraphrase[Materials and Methods, Clinical colon cancer tissues]

“The method states tissues from 30 patients but provides no demographic details (age, sex, comorbidities).”

FAILReviewer 2· 90% conf

Biological variables are poorly reported: sex, age, weight, health status, and housing conditions for mice are not described. Only species/strain/source is reported for cell lines and mice.

Evidence

direct quote[Materials and Methods, Cell culture and Mice model]

“Colon cancer cell lines (SW480, SW620 and HCT8) and normal human colon epithelial cell line (HIEC and NCM460) were obtained from ATCC (American Typical Culture Center).”

direct quote[Materials and Methods, Mice model]

“Kras G12D ;P53 f/f and Villin-Cre mice were obtained from Jacks Lab (Koch Institute for Integrative Cancer Research, Cambridge, MA).”

absence[Materials and Methods, Mice model]

Sex, age, weight, health status, and housing conditions are not reported.

For human tissues, 'this study was approved by the ethics committee of the hospital' does not name the committee. 'All of the patients agree with this study' is vague. For mice, the IACUC of Fudan University is named, which is adequate. No regulatory compliance statement is provided.

Sub-criteria

iacuc statementADEQUATE

informed consentINADEQUATE

irb ethics statementINADEQUATE

regulatory complianceNOT REPORTED

Per reviewer (2) — the votes behind the verdict

PASSReviewer 1· 90% conf

Ethics approval is reported for both human tissue collection and animal experiments, with statements of informed consent and regulatory compliance.

Evidence

direct quote[Materials and Methods, Clinical colon cancer tissues]

“This study was approved by the ethics committee of the hospital.”

direct quote[Materials and Methods, Clinical colon cancer tissues]

“All of the patients agree with this study.”

direct quote[Materials and Methods, Mice model]

“Mice were housed and treated after being approved by the Institutional Animal Care and Use Committee of Fudan University.”

WARNReviewer 2· 85% conf

Ethical approvals are mentioned but inadequately reported: the human ethics committee is not named, consent is vague, and regulatory compliance is not stated. Animal ethics approval is adequately named.

Evidence

direct quote[Materials and Methods, Clinical colon cancer tissues]

“This study was approved by the ethics committee of the hospital.”

direct quote[Materials and Methods, Clinical colon cancer tissues]

“All of the patients agree with this study.”

direct quote[Materials and Methods, Mice model]

“Mice were housed and treated after being approved by the Institutional Animal Care and Use Committee of Fudan University.”

Antibodies are listed with vendor and dilution but no catalog numbers. Cell lines are from ATCC but no authentication mentioned. Mycoplasma testing is not reported. Primer sequences are provided, but other reagents like vectors are described without catalog numbers. Software (Prism GraphPad) is named but no version.

Sub-criteria

mycoplasma testingNOT REPORTED

reagents identifiedINADEQUATE

organisms identifiedINADEQUATE

antibodies identifiedINADEQUATE

cell line authenticationNOT REPORTED

software tools identifiedINADEQUATE

Per reviewer (2) — the votes behind the verdict

WARNReviewer 1· 85% conf

Antibodies are identified with vendors and catalog information, but dilutions are not always provided; cell lines are identified by name but not authenticated; mycoplasma testing is not reported.

Evidence

direct quote[Materials and Methods, Western blot]

“anti-FOXN3 (Abcam, 1:2000), anti-Snail (Cell Signaling Technology, 1:1000), anti-Cyclin D1 (Abcam, 1:1000), anti-c-Myc and anti-GAPDH (Santa Cruz, 1:1000).”

absence[Materials and Methods, Cell culture]

Cell line authentication is not reported.

absence[Materials and Methods, Cell culture]

Mycoplasma testing is not reported.

FAILReviewer 2· 90% conf

Key resources are inadequately identified: antibodies lack catalog numbers, cell lines are not authenticated, mycoplasma testing is absent, and software lacks version numbers.

Evidence

direct quote[Materials and Methods, Western blot]

“anti-FOXN3 (Abcam, 1:2000), anti-Snail (Cell Signaling Technology, 1:1000), anti-Cyclin D1 (Abcam, 1:1000), anti-c-Myc and anti-GAPDH (Santa Cruz, 1:1000).”

direct quote[Materials and Methods, Cell culture]

“Colon cancer cell lines (SW480, SW620 and HCT8) and normal human colon epithelial cell line (HIEC and NCM460) were obtained from ATCC (American Typical Culture Center).”

direct quote[Materials and Methods, Statistical analysis]

“Statistical analyses were performed by the Student t-test (two-tailed) using Prism GraphPad software.”

The sole statistical test is the Student's t-test. No assessment of normality or variance homogeneity is provided. P-values are reported as thresholds (<0.05) rather than exact values. Effect sizes and CIs are not reported. Data are shown as bar graphs with SEM, but individual data points are not displayed.

Sub-criteria

tests namedADEQUATE

exact p valuesNOT REPORTED

effect sizes ciNOT REPORTED

data presentationINADEQUATE

software identifiedADEQUATE

assumptions verifiedNOT REPORTED

mathematical plausibilityN/A

Per reviewer (2) — the votes behind the verdict

WARNReviewer 1· 80% conf

Statistical tests are named (two-tailed Student's t-test), and software is identified (GraphPad Prism), but exact p-values are not reported, effect sizes and CIs are missing, and assumptions are not verified. Data presentation shows bar graphs without individual points.

Evidence

direct quote[Materials and Methods, Statistical analysis]

“Statistical analyses were performed by the Student t-test (two-tailed) using Prism GraphPad software.”

direct quote[Materials and Methods, Statistical analysis]

“Differences with P < 0.05 were considered statistically significant.”

direct quote[Results, Figure legends]

“Data were represented as mean±SEM.”

FAILReviewer 2· 90% conf

Statistical reporting is incomplete: only a two-tailed t-test is named, assumptions are not verified, exact p-values are not reported, effect sizes and confidence intervals are absent, and individual data points are not shown.

Evidence

direct quote[Materials and Methods, Statistical analysis]

“Statistical analyses were performed by the Student t-test (two-tailed) using Prism GraphPad software. Differences with P < 0.05 were considered statistically significant.”

direct quote[Materials and Methods, Statistical analysis]

“Data were represented as mean±SEM.”

absence[Results and Methods]

Assumptions, exact p-values, effect sizes, and CIs are not reported.

The paper does not include a data availability statement, nor does it deposit raw data in any repository. The Oncomine dataset is referenced but not directly accessible. No custom code is shared.

Sub-criteria

code sharingNOT REPORTED

accession numbersNOT REPORTED

repository depositNOT REPORTED

data availability statementNOT REPORTED

Per reviewer (2) — the votes behind the verdict

FAILReviewer 1· 90% conf

No data availability statement is provided; no data or code are deposited in any repository. The paper mentions using the Oncomine database but does not provide any identifiers for the data.

Evidence

absence[Entire paper]

No data availability statement is present.

absence[Entire paper]

No repository deposit or accession numbers are mentioned.

direct quote[Results]

“we first turned to a dataset available in the Oncomine database.”

FAILReviewer 2· 95% conf

No data or code availability statement is provided; no repository deposits or accession numbers are reported.

Evidence

absence[Entire paper]

No data availability statement is present.

absence[Entire paper]

No repository deposit or accession numbers are provided.

absence[Entire paper]

No code sharing is mentioned.

Methods cover key experimental steps, though some details (e.g., specific microscope settings for immunohistochemistry) are missing. No trial registration is needed. A limitation is briefly mentioned in the discussion regarding the lack of mechanism for FOXN3 down-regulation. Conclusions are proportional to the evidence, mainly describing associations. Funding and COI statements are present. No reporting guideline (e.g., ARRIVE) is referenced.

Sub-criteria

funding coiADEQUATE

trial registrationN/A

reporting guidelineNOT REPORTED

methods completenessADEQUATE

all outcomes reportedINADEQUATE

limitations discussedINADEQUATE

conclusions proportionalADEQUATE

Per reviewer (2) — the votes behind the verdict

WARNReviewer 1· 85% conf

Methods are detailed enough for replication, but there is no trial registration, no reporting guideline checklist, limitations are only briefly discussed, and some outcomes appear underreported. Funding and conflicts of interest are reported.

Evidence

direct quote[Discussion]

“Although we did not examine the mechanisms for the down-regulation of FOXN3 in colon cancer, several previous studies have provided some hints.”

direct quote[Acknowledgments and Footnotes]

“This study was supported by grants from Shanghai Municipal Commission of Health and Family Planning... CONFLICTS OF INTEREST There is no conflict of interest.”

absence[Entire paper]

No reporting guideline is referenced.

PASSReviewer 2· 85% conf

The paper reports methods in sufficient detail, discusses limitations, and includes a funding and conflict of interest statement. However, it does not reference a reporting guideline.

Evidence

direct quote[Materials and Methods, various sections]

“Cells were cultured in DMEM medium supplemented with 10% fetal bovine serum (GIBCO), 100units/mL penicillin and 100μg/mL streptomycin in an incubator with 5% CO2 at 37°C.”

direct quote[Discussion, paragraph 2]

“Although we did not examine the mechanisms for the down-regulation of FOXN3 in colon cancer, several previous studies have provided some hints.”

direct quote[Acknowledgments and Footnotes]

“This study was supported by grants from Shanghai Municipal Commission of Health and Family Planning (201440338)... CONFLICTS OF INTEREST There is no conflict of interest.”

14 copyedit issues flagged: mostly typo, consistency, clarity.

Checked 16 references: 15 verified — 1 not checked.

Full text (29,288 chars)Analyzed: Jul 6, 2026

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