Dear Editor,
We read with great interest the recent study by Wang et al. [
1] on the MET-TRIB3-FOXO1 regulatory axis in hepatocellular carcinoma (HCC). While the study provides valuable insights into the molecular mechanisms of HCC progression, we would like to raise several points regarding methodological rigor and interpretive clarity to ensure robust conclusions.
First, in Figure 3E, the claim that TRIB3 knockdown rescues COP1-induced FOXO1 degradation lacks critical controls. The experiment compares only COP1 overexpression + TRIB3 knockdown versus COP1 overexpression alone. However, baseline FOXO1 levels in TRIB3-knockdown cells without COP1 overexpression are missing. Without this control, the “rescue” effect may reflect TRIB3 knockdown alone altering FOXO1 stability, independent of COP1. Given TRIB3’s established roles in proteasomal degradation beyond COP1 [
2], this control is essential to validate the specificity of the interaction.
Second, study reports lung metastasis nodules using parametric tests (ANOVA), but the data distribution violates key assumptions of these tests. In the Hep3B-TRIB3 group, nodules range from 2 to 32 (mean 12.3±9.1; standard deviation >70% of mean). Such a wide range and high standard deviation relative to the mean indicate non-normal distribution. ANOVA is invalid under these conditions. Non-parametric tests (e.g., Kruskal-Wallis) are essential here. The lack of appropriate statistical analysis may exaggerate the significance of TRIB3’s role in metastasis. Reanalysis using correct statistical methods is necessary to validate these findings.
Third, study shows that ERK inhibition reduces SP1 protein levels, suggesting ERK stabilizes SP1. However, SP1 is constitutively expressed, and its degradation typically involves ubiquitination [
3]. No data demonstrate that ERK directly regulates SP1 stability. Essential experiments such as phosphorylation mutants or ubiquitination assays are missing. The observed reduction in SP1 could be off-target effects of ERK inhibition or due to general cytotoxicity rather than a specific regulatory mechanism.
Fourth, the AAV8-shTRIB3 intervention (Fig. 7) shows re-duced MET, CCND1, and TWIST1 levels. However, FOXO1 restoration alone is known to suppress these oncogenes [
4]. The study lacks rescue experiments (e.g., FOXO1 knockdown in AAV8-shTRIB3 mice) to confirm whether TRIB3’s role is specific and necessary, rather than being explained by FOXO1’s sufficiency.
In conclusion, the study by Wang et al. identifies a novel regulatory axis in HCC with potential therapeutic implications. However, several methodological and interpretive issues need rigorous validation to exclude alternative explanations and strengthen the translational relevance of the findings. Addressing these points will provide greater confidence in the proposed mechanism and its clinical significance.
FOOTNOTES
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Authors’ contributions
Bitao Jiang wrote the manuscript, Lingling Bao provided methodological and theoretical support, and Lingling Bao revised the manuscript.
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Conflicts of Interest
The authors have no conflicts to disclose.
Abbreviations
REFERENCES
- 1. Wang T, Rao D, Fu C, Sun Z, Luo Y, Lu J, et al. MET promotes hepatocellular carcinoma development through the promotion of TRIB3-mediated FOXO1 degradation. Clin Mol Hepatol 2025;31:1032-1057.
- 2. Hua F, Shang S, Yang YW, Zhang HZ, Xu TL, Yu JJ, et al. TRIB3 interacts with β-catenin and TCF4 to increase stem cell features of colorectal cancer stem cells and tumorigenesis. Gastroenterology 2019;156:708-721.e15.
- 3. Swift ML, Sell C, Azizkhan-Clifford J. DNA damage-induced degradation of Sp1 promotes cellular senescence. Geroscience 2022;44:683-698.
- 4. Yang X, Sun T, Zhao Y, Liu S, Liang X. 4sc-202 and Ink-128 cooperate to reverse the epithelial to mesenchymal transition in OSCC. Oral Dis 2022;28:2139-2148.
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