Verified Rigor RecordRigor Reviewed

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Lycium barbarum polysaccharides inhibit proliferation and migration of bladder cancer cell lines BIU87 by suppressing Pi3K/AKT pathway · 10.18632/oncotarget.13963

Name: Verified Rigor Record — Lycium barbarum polysaccharides inhibit proliferation and migration of bladder cancer cell lines BIU87 by suppressing Pi3K/AKT pathway · 10.18632/oncotarget.13963
Item: Lycium barbarum polysaccharides inhibit proliferation and migration of bladder cancer cell lines BIU87 by suppressing Pi3K/AKT pathway · 10.18632/oncotarget.13963
Rating: 62
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: highReviewer agreement: moderateEngine: 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.

The paper has a clear scientific premise and reports funding and conflicts of interest, but suffers from major deficiencies in study design (no randomization, blinding, power analysis), key resources (missing antibody catalog numbers, cell line authentication, mycoplasma testing), and data/code availability (no data statement or repository deposit). Statistical reporting is incomplete (no exact p-values, effect sizes, or assumption checks), and the paper lacks a limitations discussion and reporting guideline adherence.

Three independent reviewers evaluated all eight dimensions; their ratings converged on most dimensions, with minor divergence on scientific premise (pass vs warn) and key resources (warn vs fail), resolved by weighing the evidence. Ethical approvals was not applicable. The statistics verification component could not recompute any tests due to threshold-only p-values.

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

5 integrity concerns flagged (1 high).

Rigor

Eight Dimensions1 pass · 4 warn · 2 fail

9 major claims1 overstated1 partial7 supported

The introduction cites prior research on LBP's antitumor effects in bladder cancer cells (Mang et al.) and the PI3K/AKT pathway's role in cell proliferation and migration. It identifies a gap: 'the relationship between the PI3K/AKT signaling pathway and proliferation and migration of bladder cancer cells has not yet been reported.' The hypothesis that LBP inhibits proliferation and migration via PI3K/AKT pathway follows logically from cited evidence. Prior limitations (mechanism unclear) are acknowledged and addressed.

Sub-criteria

prior work citedADEQUATE

premise rationaleADEQUATE

limitations addressedADEQUATE

Per reviewer (3) — the votes behind the verdict

PASSReviewer 1· 90% conf

The paper adequately establishes its scientific premise by citing prior work on LBP and PI3K/AKT, providing a clear rationale, and identifying a gap in knowledge.

Evidence

direct quote[Introduction, paragraphs 2-3]

“Mang et al reported that LBP could inhibit the growth of human bladder carcinoma cell line BIU87 and induce BIU87 apoptosis []. However, the mechanism of LBP inducing apoptosis of BIU87 cells has yet been unclear.”

direct quote[Introduction, paragraph 3]

“However, the relationship between the PI3K/AKT signaling pathway and proliferation and migration of bladder cancer cells has not yet been reported.”

paraphrase[Introduction, paragraph 2]

“The paper notes the mechanism of LBP in BIU87 cells is unclear and PI3K/AKT in bladder cancer is unreported, but does not describe how these gaps are specifically mitigated in the current study.”

WARNReviewer 2· 90% conf

The paper cites prior literature on bladder cancer, LBP, and the PI3K/AKT pathway, and establishes a logical rationale for testing whether LBP acts through this pathway. However, it does not explicitly address how the limitations of prior work are mitigated in the present study.

Evidence

direct quote[Introduction, paragraph 1–2]

“LBP shows antitumor activities against various types of cancer cells and inhibit tumor growth in nude mice through induction of apoptosis and cell cycle arrest [–]. Mang et al reported that LBP could inhibit the growth of human bladder carcinoma cell line BIU87 and induce BIU87 apoptosis []. However, the mechanism of LBP inducing apoptosis of BIU87 cells has yet been unclear.”

direct quote[Introduction, paragraph 3]

“The present study was designed to study whether PI3K/AKT signaling pathway can promote the proliferation and migration of BIU87 cells, whether LBP can inhibit the proliferation and migration of BIU87cells by inhibiting PI3k/AKT pathway.”

absence[Introduction]

No limitations of prior studies are explicitly discussed as being addressed in this work.

PASSReviewer 3· 95% conf

The paper establishes a clear scientific premise by citing prior work on bladder cancer, LBP antitumor effects, and the PI3K/AKT pathway, and provides a logical rationale linking them.

Evidence

direct quote[Introduction, paragraphs 3-6]

direct quote[Introduction, final paragraph]

paraphrase[Introduction, paragraph 4 and Discussion, paragraph 5]

“The paper states that 'currently researches on the mechanism of anti-tumor effects of LBP are rare' and that this study aims to address that gap by investigating the PI3K/AKT pathway.”

Only one applicable sub-criterion (controls) is adequately reported (use of LY-294002 and IGF-1 as pathway modulators). Randomization, blinding, power analysis, explicit biological vs technical replicate distinction, inclusion/exclusion criteria, outlier handling, and independent replication are either not reported or inadequately described. This leads to a low percentage of adequate criteria.

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

FAILReviewer 1· 85% conf

The study lacks reporting of randomization, blinding, power analysis, replicate distinction, inclusion/exclusion criteria, outlier handling, and independent replication, which are critical for a rigorous preclinical in vitro study.

Evidence

direct quote[Materials and Methods, paragraph 1 (experiment reagent) and Results]

“LY-294002 is a specific inhibitor of Pi3k and can significantly inhibit the expression of p-AKT.”

absence[Materials and Methods]

No mention of randomization, blinding, or power analysis

direct quote[Materials and Methods, MTT analysis and Statistical analysis]

“All the experiments were performed for three times.”

FAILReviewer 2· 85% conf

The study design lacks critical methodological details: no randomization method, blinding, power analysis, pre-specified inclusion/exclusion criteria, or outlier handling. Replicate distinction is mentioned but the controls are inadequate for the pathway-focused design.

Evidence

absence[Materials and Methods]

No randomization method or unit is reported for cell treatment assignment.

absence[Materials and Methods]

No mention of investigator blinding at any stage of the experiment.

direct quote[Materials and Methods, Statistical analysis, and MTT analysis]

“All experiments were performed at least three times.”

paraphrase[Methods, Results]

“LY-294002 and IGF-1 are used as pathway modulators; no vehicle control for LBP is explicitly mentioned.”

WARNReviewer 3· 80% conf

The study design has several adequate elements (inclusion/exclusion, controls) but lacks critical reporting on randomization, blinding, power analysis, replicate distinction, outlier handling, and independent replication.

Evidence

direct quote[Materials and Methods, Statistical analysis; also MTT analysis, WB]

“All experiments were performed at least three times.”

direct quote[Materials and Methods, Results]

“LY-294002 is a specific inhibitor of Pi3k and can significantly inhibit the expression of p-AKT.”

absence[Methods (entire section)]

Power analysis, inclusion/exclusion criteria, and outlier handling are not mentioned.

For an in vitro study using a human cell line, only sex_reported and species_strain_source are applicable. The species and strain (BIU87) and source (Shanghai Bioleaf Co., Ltd.) are reported adequately. The sex of the BIU87 cell line is not reported (common for bladder cancer cell lines, but still a gap). The other sub-criteria (age, housing, demographics) are not applicable to cell lines.

Sub-criteria

demographicsN/A

sex reportedNOT REPORTED

sex justifiedN/A

age weight healthN/A

housing conditionsN/A

species strain sourceADEQUATE

Per reviewer (3) — the votes behind the verdict

WARNReviewer 1· 90% conf

The cell line source and species/strain are reported, but the sex of the cell line (BIU87) is not reported, and no justification is provided for using a single-sex cell line.

Evidence

direct quote[Materials and Methods, Cell culture]

“Human bladder cancer cell lines BIU87 was purchased from Shanghai Bioleaf Co., Ltd. (Shanghai, China).”

absence[Materials and Methods, Cell culture]

Sex of the cell line is not mentioned

N/AReviewer 2· 95% conf

The study uses a single human bladder cancer cell line (BIU87) and does not involve animals or human subjects, so biological variables such as sex, age, weight, and housing conditions are not applicable.

Evidence

direct quote[Materials and Methods, Cell culture]

“Human bladder cancer cell lines BIU87 was purchased from Shanghai Bioleaf Co., Ltd. (Shanghai, China).”

WARNReviewer 3· 90% conf

The paper reports the cell line source and some culture conditions but is missing critical details on sex, age/weight/health, species/strain, and housing conditions, which are largely inapplicable to an in vitro cell culture study; however, the cell line is not authenticated and mycoplasma testing is not reported.

Evidence

direct quote[Materials and Methods, Cell culture]

“Human bladder cancer cell lines BIU87 was purchased from Shanghai Bioleaf Co., Ltd. (Shanghai, China).”

absence[Methods (entire section)]

Mycoplasma testing is not reported.

direct quote[Materials and Methods, Cell culture]

“Cells were cultured at 37°C with 5% CO2 humidified atmosphere, in DMEM supplemented with 10% FBS and 1% penicillin/streptomycin.”

The research involves only an established human cancer cell line purchased from a commercial supplier, with no new human tissue, animals, or identifiable patient data. Therefore, no IRB, IACUC, or informed consent statements are applicable.

Sub-criteria

iacuc statementN/A

informed consentN/A

irb ethics statementN/A

regulatory complianceN/A

Per reviewer (3) — the votes behind the verdict

N/AReviewer 1· 100% conf

The study uses a commercially available cell line and no human subjects or animals, so ethics approval is not required.

Evidence

direct quote[Materials and Methods, Cell culture]

“Human bladder cancer cell lines BIU87 was purchased from Shanghai Bioleaf Co., Ltd.”

N/AReviewer 2· 95% conf

The study uses only a commercially purchased human cancer cell line and does not involve human subjects, animals, or non-public identifiable data; thus no ethics approval is required.

Evidence

direct quote[Materials and Methods, Cell culture]

“Human bladder cancer cell lines BIU87 was purchased from Shanghai Bioleaf Co., Ltd. (Shanghai, China).”

N/AReviewer 3· 95% conf

The study uses only a commercially available human cancer cell line (BIU87) and does not involve human or animal subjects, tissue, or non-public identifiable data; thus ethical approvals for human or animal research are not applicable.

Evidence

direct quote[Materials and Methods, Cell culture]

“Human bladder cancer cell lines BIU87 was purchased from Shanghai Bioleaf Co., Ltd. (Shanghai, China).”

absence[Entire paper]

No ethics statements are provided.

Antibodies_identified: vendors are named (Abcam, Cell Signaling Technology, Jackson ImmunoResearch, Sigma-Aldrich) but no catalog numbers, clone IDs, RRIDs, or dilution/application information are provided for any antibody — inadequate. Cell_line_authentication: the source is given but no authentication (e.g. STR profiling) is reported. Mycoplasma_testing: not reported. Organisms_identified: not applicable (only a human cell line). Reagents_identified: LBP vendor and batch number given; cell culture media, MTT, DAPI, and other reagents named with vendors; this is adequate for the materials used. Software_tools_identified: FlowJo 7.6, Image Pro Plus 6.0, and SPSS 18.0 are named with versions. Two of five applicable sub-criteria (reagents_identified, software_tools_identified) are adequate; three are not.

Sub-criteria

mycoplasma testingNOT REPORTED

reagents identifiedADEQUATE

organisms identifiedN/A

antibodies identifiedINADEQUATE

cell line authenticationNOT REPORTED

software tools identifiedADEQUATE

Per reviewer (3) — the votes behind the verdict

FAILReviewer 1· 90% conf

Key resources are inadequately identified: antibodies lack catalog numbers and validation data, cell line authentication and mycoplasma testing are not reported, and most chemical reagents lack catalog numbers, while only LBP and some software are acceptably identified.

Evidence

direct quote[Materials and Methods, Experiment reagent]

“rabbit anti-human P21, P27, MMP-2, MMP-9 polyclonal antibody(Abcam, Burlingame, CA, USA); rabbit anti-human AKT, p-AKT polyclonal antibody (Cell Signaling Technology, Inc, USA)”

absence[Materials and Methods, Cell culture]

No cell line authentication reported

absence[Materials and Methods]

No mycoplasma testing reported

WARNReviewer 2· 85% conf

Antibodies are partially identified (vendor, some catalog numbers, but no RRIDs); the cell line is identified but not authenticated; no mycoplasma testing reported; reagents are generally identified with vendors; software tools are named with versions.

Evidence

direct quote[Materials and Methods, Experiment reagent]

“rabbit anti-human P21, P27, MMP-2, MMP-9 polyclonal antibody(Abcam, Burlingame, CA, USA); rabbit anti-human AKT, p-AKT polyclonal antibody (Cell Signaling Technology, Inc, USA); horseradish peroxidase-labeled goat anti-rabbit or anti-mouse secondary antibody, FITC, FRITC labeled goat anti-rabbit IgG(Jackson Immuno Research Laboratories, Inc, USA)”

absence[Materials and Methods, Cell culture]

No cell line authentication (STR profiling) is reported.

absence[Materials and Methods]

Mycoplasma testing not reported.

direct quote[Materials and Methods, Statistical analysis]

“Statistical Package for Social Scientists (SPSS, version 18.0, IL) was used for all analyses.”

WARNReviewer 3· 85% conf

Key reagents are identified with sufficient detail (vendor, catalog/batch numbers), antibodies are identified, and software is named, but the cell line is not authenticated, mycoplasma testing is not reported, and no RRIDs are provided for resources.

Evidence

direct quote[Materials and Methods, Experiment reagent]

“rabbit anti-human P21, P27, MMP-2, MMP-9 polyclonal antibody(Abcam, Burlingame, CA, USA); rabbit anti-human AKT, p-AKT polyclonal antibody (Cell Signaling Technology, Inc, USA)”

direct quote[Materials and Methods, Experiment reagent]

“Lycium barbarum polysaccharides was purchased from Xi’an SenRan Biological Engineering Co., Ltd. (Shaanxi, China). The batch number was 20081201, and the content > 30%.”

direct quote[Materials and Methods, Wound healing; Statistical analysis]

“Cell migration area was calculated by image pro plus6.0 software ... Statistical Package for Social Scientists (SPSS, version 18.0, IL) was used for all analyses.”

Tests are adequately named and software identified. However, no exact p-values are reported (only 'P < 0.05'), assumptions (normality, equal variance) are not verified, effect sizes and confidence intervals are absent, and individual data points are not shown. Mathematical plausibility could not be assessed due to lack of numerical data. The proportion of adequate criteria is below 60%.

Sub-criteria

tests namedADEQUATE

exact p valuesINADEQUATE

effect sizes ciNOT REPORTED

data presentationINADEQUATE

software identifiedADEQUATE

assumptions verifiedNOT REPORTED

mathematical plausibilityN/A

Per reviewer (3) — the votes behind the verdict

WARNReviewer 1· 90% conf

Statistical tests are named (one-way ANOVA + LSD) and software (SPSS 18.0) is identified, but exact p-values, effect sizes, confidence intervals, and assumption checks are missing, and data presentation lacks per-group sample sizes.

Evidence

direct quote[Materials and Methods, Statistical analysis]

“Statistical analysis for multiple groups was evaluated by One-way ANOVA and LSD method.”

direct quote[Results, first paragraph]

“OD value of LBP groups was significantly lower (all P < 0.05)”

direct quote[Materials and Methods, Statistical analysis]

“Statistical Package for Social Scientists (SPSS, version 18.0, IL) was used for all analyses.”

WARNReviewer 2· 85% conf

Statistical tests are named (one-way ANOVA with LSD post-hoc), software is identified, and data presentation includes means±SD with per-group N. However, test assumptions are not verified, exact p-values are not reported (only 'p < 0.05'), effect sizes and confidence intervals are absent, and no arithmetic checks are possible from the reported summary statistics.

Evidence

direct quote[Materials and Methods, Statistical analysis]

“Statistical analysis for multiple groups was evaluated by One-way ANOVA and LSD method.”

direct quote[Results]

“OD value of LBP groups was significantly lower (all P < 0.05)”

direct quote[Materials and Methods, Statistical analysis]

“Data was presented as the means ± SD.”

WARNReviewer 3· 80% conf

The paper names the statistical test (One-way ANOVA with LSD post-hoc) and identifies software, but does not verify assumptions, report exact p-values, effect sizes, or confidence intervals, and data presentation is limited to bar graphs without individual points or defined error bars.

Evidence

direct quote[Materials and Methods, Statistical analysis]

“Data was presented as the means ± SD. ... Statistical analysis for multiple groups was evaluated by One-way ANOVA and LSD method. P < 0.05 was considered as significant. Statistical Package for Social Scientists (SPSS, version 18.0, IL) was used for all analyses.”

direct quote[Results, throughout]

“OD value of LBP groups was significantly lower (all P < 0.05)”

paraphrase[Results, Figures (not shown); Statistical analysis]

“Bar graphs and line graphs are referenced; error bars are not defined in the text but statistical analysis section mentions 'means ± SD'.”

The paper does not mention any data availability statement. Raw data (e.g., OD values, western blot images) are not deposited in any repository. No custom code is shared, and no accession numbers are provided. All four applicable sub-criteria are not reported.

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, no data deposition, no accession numbers, and no code sharing are provided, which severely limits reproducibility.

Evidence

absence[Entire paper]

No data availability statement

absence[Entire paper]

No code sharing

FAILReviewer 2· 95% conf

No data availability statement is present, no data repository deposit or accession numbers are provided, and no custom code is shared.

Evidence

absence[Entire paper]

No data availability statement is present.

absence[Entire paper]

No repository deposit or accession numbers.

absence[Entire paper]

No custom code shared.

FAILReviewer 3· 95% conf

No data availability statement, repository deposit, accession numbers, or code sharing are reported. The paper does not mention any data sharing.

Evidence

absence[Entire paper]

No data availability statement is present.

absence[Abstract, Methods, Results, Discussion, Acknowledgments]

No repository deposits, accession numbers, or code repositories are mentioned.

Methods describe the experimental procedures but lack catalog numbers for many reagents. All pre-specified outcomes are reported. Conclusions are appropriately limited to in vitro findings. Funding sources and conflicts of interest are disclosed. However, no reporting checklist (e.g., ARRIVE) is mentioned, and no limitations section is present. The proportion of adequate criteria is 50%.

Sub-criteria

funding coiADEQUATE

trial registrationN/A

reporting guidelineNOT REPORTED

methods completenessINADEQUATE

all outcomes reportedADEQUATE

limitations discussedNOT REPORTED

conclusions proportionalADEQUATE

Per reviewer (3) — the votes behind the verdict

WARNReviewer 1· 90% conf

Methods are partially complete, all outcomes are reported, conclusions are proportional, and funding/COI are stated, but no reporting guideline is followed, limitations are not discussed, and methods lack some catalog numbers for full replicability.

Evidence

direct quote[Materials and Methods, Experiment reagent]

“MTT (Sigma-Aldrich, USA); DAPI (Roche Ltd, Shanghai, China)”

paraphrase[Results sections]

“All results mentioned in Methods are presented in the Results.”

direct quote[Funding and Conflicts of Interest sections]

“This study was financially supported by Zhejiang Traditional Chinese Medicine Science and Technology Plan A (No. 2016ZA201). CONFLICTS OF INTEREST None to declare.”

WARNReviewer 2· 85% conf

Methods are generally described but lack critical details for full replication (e.g., blinding, randomization, exact antibody dilutions). A limitations discussion is absent. Conclusions are proportional to the in vitro evidence presented. Funding and COI statements are provided.

Evidence

paraphrase[Materials and Methods]

“No randomization, blinding, or antibody dilution factors are reported.”

absence[Discussion]

Limitations are not discussed.

direct quote[Footnotes]

“FUNDING This study was financially supported by Zhejiang Traditional Chinese Medicine Science and Technology Plan A (No. 2016ZA201). CONFLICTS OF INTEREST None to declare.”

WARNReviewer 3· 85% conf

Methods are fairly detailed, funding and COI are reported, but the paper lacks a trial registration (not applicable), reporting guideline (ARRIVE, not used), fails to report all outcomes (only positive results highlighted, null results ambiguous), and the conclusions are slightly overstated beyond the in vitro model.

Evidence

paraphrase[Materials and Methods]

“Detailed protocols are provided for cell culture, MTT, western blot, transwell, wound healing, and qRT-PCR, including reagent concentrations, incubation times, and analysis methods.”

direct quote[Footnotes after Acknowledgments]

“FUNDING This study was financially supported by Zhejiang Traditional Chinese Medicine Science and Technology Plan A (No. 2016ZA201). CONFLICTS OF INTEREST None to declare.”

absence[Discussion (entire section)]

Limitations are not discussed.

direct quote[Discussion, final sentence]

“Therefore, LBP may prove beneficial as a therapeutic strategy for bladder cancer treatment.”

30 copyedit issues flagged (17 major): mostly grammar, consistency, typo.

Checked 26 references: 25 verified — 1 not checked.

Αlpha¹ Rigor Action ItemsFinal pass before journal submission

This published paper has significant rigor gaps that an informed reader should weigh: the lack of randomization, blinding, and power analysis in the study design, missing key resource identifiers (antibody catalog numbers, cell line authentication), and absence of a data availability statement limit reproducibility and confidence in the findings. The implausible LY-294002 concentration (20 mmol/L, likely a unit error) and unreported qRT-PCR data are specific concerns that would warrant a correction or independent re-analysis.

1.
HIGHrigor
Correct the LY-294002 concentration from 20 mmol/L to 20 µmol/L (or justify the concentration used) in the Results section; this is likely a unit error that invalidates the interpretation of the inhibitor experiments.
A 20 mM concentration is cytotoxic and far exceeds typical effective concentrations (10-20 µM), making the results of the pathway inhibition experiments questionable.
2.
HIGHreporting
Add a data availability statement and deposit raw data (MTT OD values, western blot images, flow cytometry data) in a public repository like Figshare or Zenodo with a DOI.
Without raw data, the results cannot be independently verified or reproduced, which is a fundamental requirement for scientific reproducibility.
3.
HIGHreporting
Report the qRT-PCR data that are described in the Methods but never mentioned in the Results, or remove the qRT-PCR method if it was not performed.
A method described but not reported in the results is a serious reporting inconsistency that undermines the integrity of the paper.
4.
HIGHrigor
Perform and report STR authentication of the BIU87 cell line and state mycoplasma testing results with method and outcome in the Materials and Methods.
Without authentication and mycoplasma testing, the identity and purity of the cell line are unverified, which can invalidate all experimental results.
5.
HIGHreporting
Provide catalog numbers and RRIDs for all antibodies and key reagents (e.g., MTT, DAPI, antibodies) in the Materials section.
Missing catalog numbers and RRIDs prevent other researchers from using the same reagents, severely limiting replicability.
6.
HIGHreporting
Add a Limitations section in the Discussion addressing the in vitro nature, single cell line, lack of blinding, and absence of in vivo validation.
A limitations section is essential for readers to interpret the scope and generalizability of the findings.
7.
HIGHreporting
Temper the conclusion 'LBP may prove beneficial as a therapeutic strategy for bladder cancer treatment' to state that the findings require in vivo validation.
This conclusion overstates what a single in vitro study on one cell line can support and could mislead readers.
8.
HIGHstatistics
Report exact p-values (e.g., p=0.023) instead of inequality thresholds (P < 0.05) for all statistical comparisons.
Threshold-only p-values prevent readers from assessing the strength of evidence and are a common reporting deficiency.
9.
HIGHstatistics
Report effect sizes (e.g., Cohen's d, mean differences) and their 95% confidence intervals alongside p-values.
Effect sizes and confidence intervals provide information on the magnitude and precision of effects, which p-values alone do not.
10.
MEDIUMreporting
Describe the randomization method (e.g., random assignment of wells) and blinding (e.g., operator blinded to treatment) for the in vitro experiments in the Materials and Methods.
Randomization and blinding are essential to minimize bias in cell-based experiments, even if not always feasible.
11.
MEDIUMreporting
Report a power analysis or sample size calculation to justify the number of independent biological replicates.
Without a power analysis, it is unclear whether the study had sufficient statistical power to detect meaningful effects.
12.
MEDIUMreporting
Clarify the distinction between biological and technical replicates; state that experiments were repeated three times independently and specify the number of technical replicates per experiment.
Ambiguous replication reporting makes it impossible to assess the reliability and variability of the data.
13.
MEDIUMreporting
Pre-specify inclusion/exclusion criteria for cell passages or data points and describe outlier handling in the Materials and Methods.
Without pre-specified criteria, there is a risk of selective reporting or data manipulation.
14.
MEDIUMreporting
Follow a reporting guideline such as ARRIVE for preclinical studies and cite it in the Methods.
Adherence to reporting guidelines improves completeness and transparency of the manuscript.
15.
LOWcopyedit
Correct the spelling of 'Statistical Package for Social Scientists' to 'Statistical Package for the Social Sciences' in the Materials and Methods.
The software name is incorrect, which could confuse readers.
16.
LOWcopyedit
Fix the typo 'bolcked' to 'blocked' in the Western blot section of Materials and Methods.
Spelling errors reduce the professional quality of the manuscript.
17.
LOWcopyedit
Correct the double word typo 'Reversereverse' to 'Reverse transcription' in the Materials and Methods.
This typo could cause confusion about the method used.
18.
LOWcopyedit
Use consistent capitalization for the pathway: 'PI3K/AKT' throughout the manuscript (currently varies as Pi3K, PI3K, Pi3k).
Inconsistent capitalization of gene/protein names is unprofessional and can cause confusion.
19.
LOWcopyedit
Add a space between 'BIU87' and 'cells' throughout the manuscript (e.g., 'BIU87 cells' instead of 'BIU87cells').
Missing spaces are a formatting error that affects readability.

Full text (24,462 chars)Analyzed: Jul 6, 2026

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