Verified Rigor RecordRigor Reviewed

Rigor-Reviewed Manuscript

Circ_0081054 facilitates melanoma development via sponging miR‐637 and regulating RAB9A · 10.1111/srt.13313

Name: Verified Rigor Record — Circ_0081054 facilitates melanoma development via sponging miR‐637 and regulating RAB9A · 10.1111/srt.13313
Item: Circ_0081054 facilitates melanoma development via sponging miR‐637 and regulating RAB9A · 10.1111/srt.13313
Rating: 39
Author: Αlpha¹

Paper source

Αlpha¹ Editorial

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

Αlpha¹ Rigor Review

Rigor Reviewed

CRITICAL

Critical rigor failure · Interventional Study

Eight dimensions — click a segment for details

Critical

High-priority

Model-reported confidence: moderateReviewer agreement: highEngine: 7.5.0Source: full text

Not verified in this report

Declared data/code links were not checked for liveness or content.

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.

Critical findings

• internal_contradiction (high, corroborated): Data availability statement claims no new data were created, yet the paper reports experimental data from qPCR, Western blot, tumor measurements, etc.
• internal_contradiction (high, corroborated): The data availability statement says 'no new data were created or analyzed' but the paper presents extensive experimental data (qPCR, Western blots, flow cytometry, etc.). This is a serious internal contradiction.

The paper has a clear scientific premise and adequate reporting of ethical approvals and key resources. However, multiple methodological details are missing (randomization, blinding, power analysis, exact p-values, effect sizes, and a truthful data availability statement), and biological variables are incompletely reported (no human demographics, no justification for single-sex mouse use). The paper is already published, and these gaps reduce its robustness.

Two independent reviews (both using AI) were synthesized, with minor divergence on biological variables (warn vs. pass) resolved by weighing the evidence. The copyedit pass and verification components (citation, statistics, integrity, claim audit) were integrated. The statistics verification was limited to 1 test; all other tests reported with threshold p-values were not verifiable. The citation check identified 13 references not found in any registry, which is a potential fabrication signal.

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

Found 1 reported test, but none could be recomputed (missing degrees of freedom or sample size).

4 integrity concerns flagged (2 high).

Rigor

Eight Dimensions3 pass · 5 warn

11 major claims1 overstated1 partial9 supported

The introduction cites multiple prior studies on circRNAs in melanoma (e.g., circ_0062270, circ_0084043, circ_0016418) and discusses their roles. The rationale linking circ_0081054 to melanoma progression is clear, and the hypothesis that it acts via miR‐637/RAB9A is logically derived from prior evidence. The paper does not explicitly address limitations of prior research, but this is a minor gap.

Sub-criteria

prior work citedADEQUATE

premise rationaleADEQUATE

limitations addressedNOT REPORTED

Per reviewer (2) — the votes behind the verdict

PASSReviewer 1· 90% conf

The paper adequately establishes its scientific premise by citing prior work on circRNAs in melanoma, providing a logical rationale for studying circ_0081054, and partially addressing limitations of prior research.

Evidence

direct quote[Introduction, paragraph 2]

“Circ_0062270 regulated miR-331-3p/EPHA2 axis thus promoted melanoma development.”

direct quote[Introduction, paragraph 2]

“Circ_0081054 was upregulated in melanoma tissues, but, the role and regulatory mechanism of circ_0081054 in melanoma is still unclear.”

absence[Introduction]

The paper does not explicitly discuss how it addresses limitations of prior studies.

PASSReviewer 2· 95% conf

The paper adequately establishes the scientific premise by citing prior work on circRNAs in melanoma and stating the gap in knowledge about circ_0081054.

Evidence

direct quote[Introduction, paragraph 3]

“For example, circ_0062270 regulated miR-331-3p/EPHA2 axis thus promoted melanoma development. CircRNA_0084043 regulated miR-153-3p to promote cell proliferation and metastasis in melanoma.”

direct quote[Introduction, final paragraph]

“In current study, we intended to measure dysregulations and biological roles of circ_0081054, miR-637 and RAB9A... in melanoma tissues and cell lines, and of note the potential ceRNA network of circ_0081054/miR-637/RAB9A was planned to be further predicted and validated.”

absence[Introduction]

No explicit discussion of limitations of prior research is found.

The study uses two melanoma cell lines and includes appropriate controls (si-con, miR-con, pcDNA). Inclusion/exclusion criteria for samples are implied (melanoma tissues vs. normal skin). Replication is present for key experiments in both A375 and A2058 cells. However, randomization method for animal allocation is not reported, blinding of experimenters during cell/mouse experiments is not mentioned, no a priori power analysis is provided, and the distinction between biological and technical replicates (the paper states 'three repeats of cell experiments' but does not clarify whether these are independent biological repeats) is not adequately defined.

Sub-criteria

controlsADEQUATE

power analysisNOT REPORTED

blinding levelsNOT REPORTED

outlier handlingNOT REPORTED

randomization unitNOT REPORTED

inclusion exclusionINADEQUATE

randomization methodNOT REPORTED

replicate distinctionINADEQUATE

independent replicationADEQUATE

Per reviewer (2) — the votes behind the verdict

WARNReviewer 1· 85% conf

The study design has several strengths (controls, inclusion/exclusion criteria, replication in two cell lines) but lacks reporting on randomization method, blinding, power analysis, and distinction between biological and technical replicates.

Evidence

absence[Methods, section 2.17]

No randomization method for animal grouping is reported.

absence[Methods]

Blinding of experimenters is not mentioned.

paraphrase[Methods, section 2.18]

“The paper says 'There were three repeats of cell experiments' without specifying if these are independent biological replicates or technical replicates.”

WARNReviewer 2· 80% conf

The study design lacks reporting of randomization, blinding, power analysis, and independent replication, and the handling of outliers is not mentioned.

Evidence

absence[Methods]

No randomization method is reported.

absence[Methods]

No blinding is reported.

absence[Methods]

No a priori power analysis is reported.

direct quote[Methods, Statistical analysis]

“There were three repeats of cell experiments.”

Sex of mice is reported (male) and age (5-week-old). Species/strain/source for cell lines (HEM, A375, A2058, HUVECs) are reported with vendor/catalog numbers. For mice, only 'pathogen-free environment' is stated without details on light cycle, temperature, enrichment, or cage density. Since only male mice were used, a scientific justification for single-sex use is needed but not provided. Human demographics for the 52 melanoma patients are not reported (age, sex, race/ethnicity).

Sub-criteria

demographicsNOT REPORTED

sex reportedADEQUATE

sex justifiedNOT REPORTED

age weight healthINADEQUATE

housing conditionsINADEQUATE

species strain sourceADEQUATE

Per reviewer (2) — the votes behind the verdict

WARNReviewer 1· 90% conf

The paper reports sex and age of mice used in vivo, and provides species/strain/source for cells, but fails to report sex or justify single-sex use for cells, and does not report housing conditions for mice beyond 'pathogen-free environment'.

Evidence

direct quote[Methods, section 2.17]

“A total of 18 male BALB/C mice (male; 5-week-old)”

direct quote[Methods, section 2.17]

“were kept in pathogen-free environment”

absence[Methods, section 2.1]

Human demographics are not reported.

PASSReviewer 2· 90% conf

The paper reports sex, age, and strain for the animal experiment, and includes a demographics-relevant statement for human tissues, but human demographic details are minimal.

Evidence

direct quote[Methods, Tumor xenograft assay]

“A total of 18 male BALB/C mice (male; 5-week-old)”

direct quote[Methods, Tumor xenograft assay]

“5-week-old”

absence[Methods, Sample collection]

Human demographics (age, sex, etc.) are not reported for the tissue samples.

The paper states that the study was approved by the Ethics Committee of the Affiliated Huaian NO.1 People's Hospital of Nanjing Medical University, and that all patients provided informed consent. The animal study was approved by the Animal Care and Use Committee of the same institution. Regulatory compliance is implied but not explicitly stated (e.g., Declaration of Helsinki).

Sub-criteria

iacuc statementADEQUATE

informed consentADEQUATE

irb ethics statementADEQUATE

regulatory complianceNOT REPORTED

Per reviewer (2) — the votes behind the verdict

PASSReviewer 1· 95% conf

Ethical approvals are adequately reported for both human sample collection and animal experiments, with named ethics committees and informed consent for patients.

Evidence

direct quote[Methods, section 2.1]

“This research was conducted with the approval of the Ethics Committee of the Affiliated Huaian NO.1 People's Hospital of Nanjing Medical University.”

direct quote[Methods, section 2.1]

“All patients provided informed consent before starting this study.”

direct quote[Methods, section 2.17]

“Animal study was approved by the Animal Care and Use Committee of the Affiliated Huaian NO.1 People's Hospital of Nanjing Medical University.”

PASSReviewer 2· 95% conf

Ethical approvals for both human tissue collection and animal experiments are reported with named committees and consent documentation.

Evidence

direct quote[Methods, Sample collection]

“This research was conducted with the approval of the Ethics Committee of the Affiliated Huaian NO.1 People's Hospital of Nanjing Medical University.”

direct quote[Methods, Sample collection]

“All patients provided informed consent before starting this study.”

direct quote[Methods, Tumor xenograft assay]

“Animal study was approved by the Animal Care and Use Committee of the Affiliated Huaian NO.1 People's Hospital of Nanjing Medical University.”

Cell lines (HEM, A375, A2058) are identified with vendor and catalog numbers. Antibodies are identified with catalog numbers and dilutions (e.g., HK2 ab227198 1:1000). Software (SPSS 22.0, ImageJ) is named with versions. Primer sequences are provided in a table. The investigational product is not a drug but a biological manipulation (siRNA/plasmids), which is adequately identified. Mycoplasma testing is not reported.

Sub-criteria

mycoplasma testingNOT REPORTED

reagents identifiedADEQUATE

organisms identifiedADEQUATE

antibodies identifiedADEQUATE

cell line authenticationADEQUATE

software tools identifiedADEQUATE

Per reviewer (2) — the votes behind the verdict

PASSReviewer 1· 95% conf

Key resources are well-identified, including cell lines with sources, antibodies with catalog numbers and dilutions, and software with versions.

Evidence

direct quote[Methods, section 2.11]

“hexokinase II (HK2; ab227198, 1:1000), β-actin (ab8227, 1:5,000)”

direct quote[Methods, section 2.2]

“Melanoma cell lines A375 and A2058 were bought from American Type Culture Collection (ATCC, Manassas, VA, USA).”

absence[Methods]

Mycoplasma testing is not reported.

PASSReviewer 2· 85% conf

Key resources including cell lines, antibodies, reagents, and software are identified with sources, catalog numbers, and RRIDs where applicable.

Evidence

direct quote[Methods, Cell lines]

“Human epidermal melanocyte (HEM) was bought from ScienCell (Carlsbad, CA, USA)... Melanoma cell lines A375 and A2058 were bought from American Type Culture Collection (ATCC, Manassas, VA, USA).”

direct quote[Methods, Western blot analysis]

“the antibodies (Abcam, Cambridge, UK) including: hexokinase II (HK2; ab227198, 1:1000), β-actin (ab8227, 1:5,000), Goat Anti-Rabbit IgG H&L secondary antibody (ab205718, 1:3000), RAB9A (ab235538, 1:2000).”

direct quote[Methods, Statistical analysis]

“SPSS 22.0 (SPSS Inc., Chicago, IL, USA)”

The paper states use of Student's t-test or ANOVA, and SPSS 22.0. However, all p-values are reported only as '* p < 0.05' without exact values. No effect sizes or confidence intervals are reported. Data are presented as bar graphs with error bars (likely SD based on 'mean ± standard deviation') but individual data points are not shown. The paper does not mention verification of test assumptions (normality, equal variance). Mathematical plausibility checks are not applicable given the presentation style (no raw means with Ns to check).

Sub-criteria

tests namedADEQUATE

exact p valuesINADEQUATE

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 and software identified, but exact p-values are not reported (only '* p < 0.05'), effect sizes and confidence intervals are absent, and data are presented as bar graphs without individual points.

Evidence

direct quote[Results (all figures)]

“* p < 0.05”

absence[Methods, section 2.18]

Test assumptions are not mentioned.

direct quote[Methods, section 2.18]

“SPSS 22.0 (SPSS Inc., Chicago, IL, USA)”

WARNReviewer 2· 80% conf

Statistical tests are named, but assumptions are not verified, exact p-values are mostly not reported (only 'p < 0.05'), effect sizes/CI are absent, and data presentation lacks individual data points.

Evidence

direct quote[Methods, Statistical analysis]

“Analysis of variance or Student's t-test was used to analyze relevant data through SPSS 22.0.”

direct quote[Figures and Methods]

“*p < 0.05”

absence[Results and Methods]

Effect sizes and confidence intervals are not reported.

The paper states 'Data sharing is not applicable to this article as no new data were created or analyzed in this study', which is contradictory because new experimental data (qPCR, Western blot, tumor measurements) were generated. This statement is also vague and does not provide a concrete access route. No repository deposit, accession numbers, or code sharing are reported.

Sub-criteria

code sharingNOT REPORTED

accession numbersNOT REPORTED

repository depositNOT REPORTED

data availability statementINADEQUATE

Per reviewer (2) — the votes behind the verdict

WARNReviewer 1· 95% conf

A data availability statement is present but is vague and contradictory ('data sharing not applicable as no new data were created'), despite the paper clearly generating new data. No code is shared.

Evidence

direct quote[Data Availability Statement]

“Data sharing is not applicable to this article as no new data were created or analyzed in this study.”

absence[Data Availability Statement / Methods]

No code repository is mentioned.

WARNReviewer 2· 90% conf

The data availability statement is provided but is vague ('available on request' is implied but not present; rather it says 'not applicable as no new data were created'), which is inconsistent with the study's own data generation.

Evidence

direct quote[Data Availability Statement, end of paper]

“Data sharing is not applicable to this article as no new data were created or analyzed in this study.”

absence[Data Availability Statement]

No repository deposit is reported.

Methods are described in sufficient detail to allow replication. Limitations are discussed (e.g., need for further in vivo validation of miR-637/RAB9A). Conclusions are proportional to the correlative and in vitro evidence. However, no reporting guideline (ARRIVE, CONSORT) is referenced. Funding statement says 'no specific funding' and conflicts of interest are declared. The contradictory data availability statement is a transparency issue.

Sub-criteria

funding coiADEQUATE

trial registrationN/A

reporting guidelineNOT REPORTED

methods completenessADEQUATE

all outcomes reportedADEQUATE

limitations discussedADEQUATE

conclusions proportionalADEQUATE

Per reviewer (2) — the votes behind the verdict

WARNReviewer 1· 85% conf

Methods are relatively complete for replication, limitations are discussed, and conclusions are proportional. However, the paper does not follow any specific reporting guideline, and the data availability statement is contradictory.

Evidence

absence[Methods]

No reporting guideline is mentioned.

direct quote[Discussion]

“Further investigation should focus on the function of miR-637 and RAB9A on tumor formation in vivo.”

direct quote[Funding Information / Conflict of Interest Statement]

“The authors declare that they have no conflict of interest.”

WARNReviewer 2· 85% conf

Methods are generally complete for replication, but no trial registration, reporting guideline, or pre-specified outcome protocol is present, and limitations are discussed but limited.

Evidence

paraphrase[Methods sections 2.2–2.18]

“All cell culture, transfection, and assay protocols are described in sufficient detail for replication.”

absence[Methods]

No reporting guideline is mentioned.

direct quote[Discussion, final paragraph]

“However, the in vivo experiment only investigated the expression of miR-637 and RAB9A in the primary tumors from A375 cells. Further investigation should focus on the function of miR-637 and RAB9A on tumor formation in vivo.”

14 copyedit issues flagged (1 major): mostly typo, clarity, consistency.

Checked 28 references: 15 verified — 13 unresolved.

Αlpha¹ Rigor Action ItemsFinal pass before journal submission

As a post-publication audit, the paper's robustness is compromised by the contradictory data availability statement, missing human demographics, incomplete statistical reporting, and the large number of references not found in registries. An informed reader should weigh these issues heavily; the paper would warrant an erratum to correct the data availability statement and a careful independent re-analysis of the reported statistics. The missing details on randomization, blinding, and power analysis limit the interpretability of the in vivo results.

1.
CRITICALdata code
Replace the contradictory data availability statement with one that accurately reflects the generated data, e.g., 'The data that support the findings of this study are available from the corresponding author upon reasonable request' or deposit raw data in a public repository.
The current statement claims no new data were created, which is false and undermines trust; this must be corrected via an erratum.
2.
CRITICALrigor
Verify the 13 references not found in any registry: "Malignant melanoma: skin cancer‐diagnosis, prevention, and treatment", "Skin cancer", "Malignant melanoma: beyond the basics", "Screening for reducing morbidity and mortality in malignant melanoma", "Circular RNAs: identification, biogenesis and function", "The potential role of miR‐29 in health and cancer diagnosis, prognosis, and therapy", "miR‐146a‐5p: expression, regulation, and functions in cancer", "Association of miR‐760 with cancer: an overview", "MiR‐320a inhibits malignant phenotype of melanoma cells via targeting PBX3", "miR‐509‐3p suppresses migration, invasion, and epithelial‐ mesenchymal transition in melanoma cells by targeting collagen triple helix repeat containing 1", "Knockdown of circ_0084043 suppresses the development of human melanoma cells through miR‐429/tribbles homolog 2 axis and Wnt/beta‐catenin pathway", "Knockdown of Rab9 suppresses the progression of gastric cancer through regulation of Akt signaling pathway". Provide correct references or remove them.
References not found in registries are a potential fabrication signal and must be resolved for scientific integrity.
3.
HIGHreporting
Temper the claim in the Discussion that "Circ_0081054 could be a promising target to combat the development of melanoma" to reflect the early preclinical evidence (cell lines and a single xenograft model) and acknowledge the need for further validation.
The claim is overstated given the limited evidence; over-claiming is a common reviewer objection and can mislead readers.
4.
HIGHstatistics
Report exact p-values (e.g., p = 0.023) instead of only asterisks in all figures and tables, and provide effect sizes (e.g., Cohen's d) and 95% confidence intervals for key comparisons.
Threshold-only p-values are insufficient for readers to assess the strength of evidence; exact values and effect sizes are standard requirements.
5.
HIGHrigor
Add a statement describing how sample size was determined (power analysis) for the in vivo xenograft experiment in Methods 2.17.
Without a power analysis, the sample size of 18 mice may be arbitrary, affecting the reliability of the conclusions.
6.
HIGHrigor
Describe the method of randomization for allocating mice to groups in the tumor xenograft assay (Methods 2.17).
Randomization is essential to avoid selection bias; its absence is a major methodological gap.
7.
HIGHrigor
State whether the experimenter was blinded during cell assays and tumor measurements, or provide a rationale if blinding was not feasible.
Blinding prevents observer bias, especially in subjective assays like colony counting and tumor measurements.
8.
HIGHrigor
Provide a justification for using only male mice in the xenograft study (Methods 2.17).
Single-sex studies require a scientific rationale to avoid excluding potential sex differences.
9.
HIGHrigor
Report the demographic characteristics (age, sex) of the 52 melanoma patients in Methods 2.1 or Results.
Missing demographic data limits the generalizability of the findings and is a basic reporting requirement.
10.
HIGHreporting
Add a statement on cell line authentication (STR profiling) and mycoplasma testing, including the method and result, to the Methods section.
Unvalidated cell lines are a common source of irreproducibility; journals increasingly require this information.
11.
MEDIUMreporting
Clarify whether the 'three repeats of cell experiments' are independent biological replicates or technical replicates in Methods 2.18.
The distinction affects the statistical analysis and interpretation of the results.
12.
MEDIUMreporting
Provide details on housing conditions for mice (light cycle, temperature, enrichment) beyond 'pathogen-free environment' in Methods 2.17.
Adequate reporting of housing conditions is necessary for reproducibility of animal experiments.
13.
MEDIUMstatistics
Include a statement on how outliers were handled in the statistical analysis, or explicitly state that no outliers were excluded.
Readers need to know whether data were excluded and why, to assess potential bias.
14.
MEDIUMreporting
Follow the ARRIVE guidelines for reporting animal experiments and include a completed checklist as supplementary material.
Many journals require ARRIVE compliance for animal studies; its absence is a transparency gap.
15.
MEDIUMreporting
Add a statement about the use of reporting guidelines (e.g., ARRIVE) in the Methods.
Referencing reporting guidelines improves completeness and reproducibility.
16.
MEDIUMcopyedit
Polish language and formatting throughout: fix grammar (e.g., 'confronting problem of humans over the worldwide'), typos (duplicate 'cells' in Figure 4 legend), consistency (Western blot vs. Western blotting), and clarify ambiguous phrases (e.g., 'the used Dual-luciferase Reporter System').
Grammatical errors and inconsistencies reduce professionalism and readability.
17.
LOWother
Clarify the inconsistency between 'Pearson correlation coefficient' and 'Pearson's rank correlation coefficient' in the Methods and Figure 7 legend; ensure correct terminology.
Mixing Pearson and rank correlation terminology is confusing and may indicate a methodological error.

Full text (40,233 chars)Analyzed: Jul 6, 2026

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