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Cited by 2Cholangiocarcinoma and Gallbladder Cancer StudiesPediatric Hepatobiliary Diseases and TreatmentsHippo pathway signaling and YAP/TAZ

Overexpression of TBX3 suppresses tumorigenesis in experimental and human cholangiocarcinoma

Open access
Deng, Shanshan, Lu, Xinjun, Wang, Xue, Liang, Binyong, Xu, Hongwei, Yang, Doris, Cui, Guofei, Yonemura, Andrew +9 more·2024·Cell Death & Disease
DOI 10.1038/s41419-024-06839-8PMC PMC11193761
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Abstract

TBX3 behaves as a tumor suppressor or oncoprotein across cancer. However, TBX3 function remains undetermined in intrahepatic cholangiocarcinoma (iCCA), a deadly primary liver malignancy with few systemic treatment options. This study sought to investigate the impact of TBX3 on iCCA. We found that overexpression of TBX3 strongly inhibited human iCCA cell growth. In the Akt/FBXW7ΔF mouse iCCA model, overexpression of Tbx3 reduced cholangiocarcinogenesis in vivo, while inducible genetic knockout of Tbx3 accelerated iCCA growth. RNA-seq identified MAD2L1 as a downregulated gene in TBX3-overexpressing cells, and ChIP confirmed that TBX3 binds to the MAD2L1 promoter. CRISPR-mediated knockdown of Mad2l1 significantly reduced the growth of two iCCA models in vivo. Finally, we found that TBX3 expression is upregulated in ~20% of human iCCA samples, and its high expression is associated with less proliferation and better survival. MAD2L1 expression is upregulated in most human iCCA samples and negatively correlated with TBX3 expression. Altogether, our findings suggest that overexpression of TBX3 suppresses CCA progression via repressing MAD2L1 expression.

AI snapshot

Written from the paper’s full text.

TL;DR

In experimental and human intrahepatic cholangiocarcinoma (iCCA), TBX3 overexpression suppresses tumorigenesis, while Tbx3 loss accelerates it. TBX3 directly binds the MAD2L1 promoter and represses MAD2L1, a gene whose deletion also inhibits iCCA growth in mouse models.

Key findings

  • TBX3 overexpression inhibited colony formation in RBE, KKU-156, and HUCCT1 human iCCA cells. In Akt/FBXW7ΔF mice, Akt/FBXW7ΔF/TBX3 mice showed no abdominal mass and were healthy at ~25 weeks, whereas all control Akt/FBXW7ΔF/pT3 mice required euthanasia between 8 and 14 weeks; liver weight and liver/body ratio were significantly lower in the TBX3 group (p < 0.0001).
  • Inducible Tbx3 knockout in Tbx3flox/flox mice injected with Akt/FBXW7ΔF/Cre increased liver weight versus Akt/FBXW7ΔF/pCMV controls (p < 0.05) and increased number/size of iCCA lesions.
  • RNA-seq of TBX3-overexpressing KKU-156 cells identified 827 upregulated and 904 downregulated genes. MAD2L1 was consistently downregulated by TBX3 in iCCA cells and mouse lesions; ChIP confirmed TBX3 binding to the MAD2L1 promoter. Mad2l1 deletion reduced tumor burden in Akt/FBXW7ΔF and Akt/NICD mouse models.
  • In human samples, TBX3 was upregulated in ~24% of NCI iCCA samples and 16/82 (19.5%) by IHC; high TBX3 was associated with longer survival (median 29.0 vs 17.5 months, p < 0.001) and lower Ki-67. MAD2L1 was upregulated in ~84% of NCI samples and 68/82 (82.3%) by IHC, negatively correlated with TBX3. In 3/10 Akt/FBXW7ΔF/TBX3 mice, mesenchymal osteosarcoma-like lesions with osteoid differentiation developed.

Methods

  • In vitro: human iCCA cell lines RBE, KKU-156, HUCCT1; TBX3 or MAD2L1 overexpression and CRISPR-Cas9 knockout; colony formation, western blot, RT-qPCR, RNA-seq, ChIP, and transcription factor binding site prediction.
  • Mouse models: hydrodynamic tail vein injection in FVB/N or Tbx3flox/flox mice using Akt/FBXW7ΔF with TBX3 overexpression or Cre-mediated Tbx3 knockout, and Akt/FBXW7ΔF or Akt/NICD with sgMad2l1; liver weight, survival, H&E, and IHC were assessed.
  • Human data: iCCA and surrounding non-tumorous liver tissues (n = 82 each) from the University of Regensburg; TCGA and NCI iCCA datasets. Analyses included Mann–Whitney U, unpaired t tests, Kaplan–Meier/log-rank, and chi-square tests; p < 0.05 was significant.

Limitations

  • Stated in the paper: the effects of Tbx3 ablation were relatively moderate, likely because Tbx3 levels are already low in Akt/FBXW7ΔF mouse iCCA lesions; why only a subset of Akt/FBXW7ΔF/TBX3 mice developed mesenchymal/osteosarcoma-like tumors is unclear; the TBX3 partner(s) driving this differentiation remain unidentified; no small molecules against MAD2L1 have been reported; additional studies are needed to determine how the TBX3/MAD2L1 pathway regulates drug responsiveness.
  • Evident from the text: human IHC analyses used a single-center cohort of 82 iCCA cases, and TCGA/NCI analyses rely on existing datasets; the mouse models use specific oncogenic drivers (Akt/FBXW7ΔF, Akt/NICD) and may not capture the full heterogeneity of human iCCA.

Model-written summary. Check it against the paper before relying on it.

Integrity signals

Registry

Crossref · 10.1038/s41419-024-06839-8

Cites

33 of 33 references verified0 citation errors · 0 not checkable

Statistics

Nothing to recompute.

Claims the paper makes

15 of 27 cited sources checked · 10 supported · 1 flagged · accuracy not yet measured

  • Premisecitation

    The Akt/FBXW7ΔF mouse model develops iCCA lesions by ~7 weeks post-injection.

    “In this model, iCCA lesions develop by ~7 weeks post-injection [30].”
    Quote in the paperNot a substantial claim1 of 1 sources checked
    • cites [30] Wang J, Wang H, Peters M, Ding N, Ribback S, Utpatel K, et al. Loss of Fbxw7 synergizes with activated Akt signaling to promote c-Myc dependent cholangiocarcinogenesis. J Hepatol. 2019;71:742–52. source
  • Premisecitation

    The Akt/FBXW7ΔF mouse model is highly relevant to human cholangiocarcinogenesis.

    “This model was chosen as we have demonstrated that it is highly relevant to human cholangiocarcinogesis [30].”
    Quote in the paperSubstantial claim1 of 1 sources checked
    • cites [30] Wang J, Wang H, Peters M, Ding N, Ribback S, Utpatel K, et al. Loss of Fbxw7 synergizes with activated Akt signaling to promote c-Myc dependent cholangiocarcinogenesis. J Hepatol. 2019;71:742–52. source
  • Premisecitation

    TBX3 is a transcriptional repressor.

    “Previous studies have shown that TBX3 is a transcriptional repressor [16].”
    • Also stated in: § MAD2L1 is a TBX3 target gene in iCCA · boosted
      “Since TBX3 is a well-characterized transcriptional repressor, we focused on the genes downregulated by TBX3 overexpression.”
    Quote in the paperSubstantial claim1 of 1 sources checked
    • cites [16] Khan SF, Damerell V, Omar R, Du Toit M, Khan M, Maranyane HM, et al. The roles and regulation of TBX3 in development and disease. Gene. 2020;726:144223. source
  • Premisecitation

    The Akt/FBXW7ΔF model was induced by co-expression of activated Akt and dominant negative FBXW7 via hydrodynamic tail vein gene injection.

    “we applied a previously established murine iCCA model, induced by the co-expression of an activated form of Akt (myr-Akt) and the dominant negative form of FBXW7 (FBXW7ΔF) (Akt/FBXW7ΔF) in the mouse liver via hydrodynamic tail vein gene injection (HTVi) [30].”
    Quote in the paperSubstantial claim1 of 1 sources checked
    • cites [30] Wang J, Wang H, Peters M, Ding N, Ribback S, Utpatel K, et al. Loss of Fbxw7 synergizes with activated Akt signaling to promote c-Myc dependent cholangiocarcinogenesis. J Hepatol. 2019;71:742–52. source
  • Findingown data

    Tbx3 protein levels are low in Akt/FBXW7ΔF iCCA lesions compared to normal livers of uninjected mice.

    “Tbx3 protein levels were low in the lesions from this model compared to normal livers of uninjected mice (Fig. 2A)”
    Quote in the paperSubstantial claim
  • Classified as probably not a substantial claim. 19
  • May combine two claims. 16

  • Probably not a substantial claim: may not pass S3 (load-bearing).

    Premisecitation

    TBX3 acts as a tumor suppressor in β-catenin-driven HCC.

    “However, TBX3 acts as a tumor suppressor in other tumor entities, including β-catenin-driven HCC [18, 21, 22].”
    Quote in the paperNot a substantial claim2 of 3 sources checked
    • cites [18] Willmer T, Cooper A, Peres J, Omar R, Prince S. The T-Box transcription factor 3 in development and cancer. Biosci Trends. 2017;11:254–66. source

      Section 2.4.1 says that in β-catenin-driven liver tumours, TBX3 promotes proliferation, tumour formation and survival—the opposite of a tumour-suppressor role. Section 2.3.4 describes a tumour-suppressor role in transformed fibroblast and fibrosarcoma models, not in β-catenin-driven HCC. (LLM, conf=0.91)

      Checked by a reasoning model — may be wrong.

      Experimental — accuracy not yet measured
    • cites [21] Liang B, Zhou Y, Qian M, Xu M, Wang J, Zhang Y, et al. TBX3 functions as a tumor suppressor downstream of activated CTNNB1 mutants during hepatocarcinogenesis. J Hepatol. 2021;75:120–31. source
    • cites [22] Renard CA, Labalette C, Armengol C, Cougot D, Wei Y, Cairo S, et al. Tbx3 is a downstream target of the Wnt/beta-catenin pathway and a critical mediator of beta-catenin survival functions in liver cancer. Cancer Res. 2007;67:901–10. source

  • Probably not a substantial claim: may not pass S3 (load-bearing). May combine two claims.

    Premisecitation

    TBX3 is a member of the T-box gene family and functions as a transcription factor.

    “TBX3 is a member of the T-box gene family and functions as a transcription factor [16].”
    Quote in the paperNot a substantial claim1 of 1 sources checked
    • cites [16] Khan SF, Damerell V, Omar R, Du Toit M, Khan M, Maranyane HM, et al. The roles and regulation of TBX3 in development and disease. Gene. 2020;726:144223. source

  • Probably not a substantial claim: may not pass S3 (load-bearing), S4 (based).

    Premisecitation

    TBX3 behaves as a tumor suppressor or oncoprotein across cancer.

    “TBX3 behaves as a tumor suppressor or oncoprotein across cancer.”
    Quote in the paperNot a substantial claim3 of 5 sources checked
    • cites [18] Willmer T, Cooper A, Peres J, Omar R, Prince S. The T-Box transcription factor 3 in development and cancer. Biosci Trends. 2017;11:254–66. source
    • cites [19] Omar R, Cooper A, Maranyane HM, Zerbini L, Prince S. COL1A2 is a TBX3 target that mediates its impact on fibrosarcoma and chondrosarcoma cell migration. Cancer Lett. 2019;459:227–39. source
    • cites [20] Aliwaini, Lubbad S, Shourfa AM, Hamada HAA A, Ayesh, Abu Tayem HEM B, et al. Overexpression of TBX3 transcription factor as a potential diagnostic marker for breast cancer. Mol Clin Oncol. 2019;10:105–12. source

      The Introduction says TBX3 overexpression may drive cancer and describes its upregulation in melanoma and breast cancer. The Discussion also links TBX3 to oncogenic processes and lists other carcinomas. Neither passage says TBX3 acts as a tumor suppressor, so the claimed alternative role is unsupported. (LLM, conf=0.86)

      Experimental — accuracy not yet measured
    • cites [21] Liang B, Zhou Y, Qian M, Xu M, Wang J, Zhang Y, et al. TBX3 functions as a tumor suppressor downstream of activated CTNNB1 mutants during hepatocarcinogenesis. J Hepatol. 2021;75:120–31. source
    • cites [22] Renard CA, Labalette C, Armengol C, Cougot D, Wei Y, Cairo S, et al. Tbx3 is a downstream target of the Wnt/beta-catenin pathway and a critical mediator of beta-catenin survival functions in liver cancer. Cancer Res. 2007;67:901–10. source

  • Probably not a substantial claim: may not pass S3 (load-bearing).

    Premisecitation

    Chemotherapy, consisting of Gemcitabine and Cisplatin-based therapies, is still the mainstream treatment option for advanced, unresectable, or recurrent iCCA.

    “Chemotherapy, consisting of Gemcitabine and Cisplatin-based therapies, is still the mainstream treatment option for advanced, unresectable, or recurrent iCCA [33].”
    Quote in the paperNot a substantial claim1 of 1 sources checked
    • cites [33] Khosla D, Misra S, Chu PL, Guan P, Nada R, Gupta R, et al. Cholangiocarcinoma: recent advances in molecular pathobiology and therapeutic approaches. Cancers. 2024;16. source

  • Probably not a substantial claim: may not pass S3 (load-bearing).

    Premisecitation

    An opposite, pro-oncogenic role for TBX3 has been revealed in other tumor types, such as malignant melanoma, breast, gastric, and cervical cancers.

    “An opposite, pro-oncogenic role for TBX3 has been revealed in other tumor types, such as malignant melanoma, breast, gastric, and cervical cancers [1–3].”
    Quote in the paperNot a substantial claim2 of 3 sources checked
    • cites [1] Rizvi S, Khan SA, Hallemeier CL, Kelley RK, Gores GJ. Cholangiocarcinoma—evolving concepts and therapeutic strategies. Nat Rev Clin Oncol. 2018;15:95–111. source

      Our copy discusses genetic drivers and therapies for cholangiocarcinoma (Abstract, page unavailable), but the passages do not address a pro-oncogenic role for TBX3 in melanoma, breast, gastric, or cervical cancer. Searches for TBX3 and its expanded name with those cancer types found no relevant passage. (LLM, conf=0.76)

      Experimental — accuracy not yet measured
    • cites [2] Siegel RL, Miller KD, Fuchs HE, Jemal A. Cancer statistics, 2022. CA Cancer J Clin. 2022;72:7–33. source
    • cites [3] Yi X, Zhu J, Liu W, Peng L, Lu C, Sun P, et al. Proteome landscapes of human hepatocellular carcinoma and intrahepatic cholangiocarcinoma. Mol Cell Proteomics. 2023:100604. source

      The Results and Discussion section examines protein expression in hepatocellular carcinoma and cholangiocarcinoma, not TBX3’s role in the cancers named in the claim. Searches for TBX3 and for an oncogenic role in melanoma, breast, gastric, and cervical cancers found no passage addressing that point in our copy. (LLM, conf=0.76)

      Experimental — accuracy not yet measured

  • Classified as probably not a substantial claim.

    Premisecitation

    Most patients do not respond to the chemotherapy.

    “However, most patients do not respond to the chemotherapy [33].”
    Quote in the paperSubstantial claim1 of 1 sources checked
    • cites [33] Khosla D, Misra S, Chu PL, Guan P, Nada R, Gupta R, et al. Cholangiocarcinoma: recent advances in molecular pathobiology and therapeutic approaches. Cancers. 2024;16. source

      The Introduction (page unavailable) says cholangiocarcinoma’s tumor characteristics and therapeutic resistance have led to refractoriness to chemotherapy. That supports poor chemotherapy response, but the paper does not establish that most patients fail to respond; §4.2 instead reports benefits from some regimens. (LLM, conf=0.82)

      Experimental — accuracy not yet measured

  • Probably not a substantial claim: may not pass S3 (load-bearing). May combine two claims.

    Premisecitation

    TBX3 expression was associated with less aggressive biological behavior and a more differentiated phenotype in a β-catenin-driven HCC model.

    “Similarly, we previously showed that TBX3 expression was associated with less aggressive biological behavior and a more differentiated phenotype in a β-catenin-driven HCC model [21].”
    Quote in the paperNot a substantial claim1 of 1 sources checked
    • cites [21] Liang B, Zhou Y, Qian M, Xu M, Wang J, Zhang Y, et al. TBX3 functions as a tumor suppressor downstream of activated CTNNB1 mutants during hepatocarcinogenesis. J Hepatol. 2021;75:120–31. source

  • May combine two claims.

    Findingown data

    High TBX3 expression is associated with less proliferation and better survival in human iCCA.

    “its high expression is associated with less proliferation and better survival.”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Findingown data

    All Akt/FBXW7ΔF/pT3 injected mice developed a considerable liver tumor burden and required euthanasia between 8 and 14 weeks post-injection.

    “All Akt/FBXW7ΔF/pT3 injected mice developed a considerable liver tumor burden and required euthanasia between 8 and 14 weeks post-injection.”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Findingown data

    The mesenchymal cells in these lesions were positive for SATB2 and RANKL.

    “The positive immunoreactivity of the mesenchymal cells for the SATB2 transcription factor and the RANK ligand (RANKL) further substantiated the presumable osteoid nature of these lesions.”
    Quote in the paperNot a substantial claim

  • May combine two claims.

    Findingown data

    All tumor iCCA lesions in both cohorts were positive for p-AKT and CK19 and negative for HNF4α.

    “all tumor iCCA lesions in both cohorts were positive for p-AKT and the biliary epithelial marker CK19 and negative for the HNF4α hepatocyte marker (Fig. 3B).”
    Quote in the paperNot a substantial claim

  • May combine two claims.

    Findingown data

    MAD2L1 immunoreactivity was moderate in the cytoplasm of hepatocytes and absent in cholangiocytes.

    “As concerns MAD2L1, immunoreactivity was moderate in the cytoplasm of hepatocytes and absent in cholangiocytes (Supplementary Fig. 10).”
    Quote in the paperNot a substantial claim

  • May combine two claims.

    Findingown data

    We identified 4761 TBX3 positively correlated genes and 2475 TBX3 negatively correlated genes.

    “We identified 4761 TBX3 positively correlated genes and 2475 TBX3 negatively correlated genes.”
    Quote in the paperSubstantial claim

  • Probably not a substantial claim: may not pass S3 (load-bearing).

    Findingown data

    The osteoid substance was demarcated by multinucleated cells resembling osteoclasts.

    “the osteoid substance was demarcated by multinucleated cells resembling osteoclasts.”
    Quote in the paperNot a substantial claim

  • May combine two claims.

    Findingown data

    RBE and KKU-156 cells display elevated protein levels of TBX3, whereas TBX3 was low in HUCCT1 cells.

    “RBE and KKU-156 cells display elevated protein levels of TBX3, whereas TBX3 was low in HUCCT1 cells (Fig. 1A).”
    Quote in the paperNot a substantial claim

  • Probably not a substantial claim: may not pass S4 (based). May combine two claims.

    Findingnone given

    Most iCCA samples are TBX3 low and MAD2L1 high.

    “As most iCCA samples are TBX3 low and MAD2L1 high”
    Quote in the paperNot a substantial claim

  • May combine two claims.

    Study design and conductown data

    Western blot analysis and Sanger sequencing confirmed the efficient deletion of the MAD2L1 gene in the iCCA cells.

    “Western blot analysis and Sanger sequencing confirmed the efficient deletion of the MAD2L1 gene in the iCCA cells (Supplementary Fig. 5B, C).”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Study design and conductown data

    TBX3 was identified as a top-upregulated gene, confirming its successful overexpression in the transfected cells.

    “TBX3 was identified as a top-upregulated gene, confirming its successful overexpression in the transfected cells (Supplementary Table 3 and Fig. 5A)”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Interpretationown data

    TBX3 directly binds to the MAD2L1 promoter and represses its expression in iCCA cells.

    “The data indicate that TBX3 directly binds to the MAD2L1 promoter and represses its expression in iCCA cells.”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Interpretationown data

    Overexpression of TBX3 or deletion of MAD2L1 leads to iCCA growth inhibition.

    “Overexpression of TBX3 or deletion of MAD2L1 leads to iCCA growth inhibition”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Premisecitation

    The expression of MAD2L1 was studied in resected iCCA specimens and correlated with tumor size, pathological grade, clinical stage, and inversely with overall survival.

    “The expression of MAD2L1 was studied in resected iCCA specimens and correlated with tumor size, pathological grade, clinical stage, and inversely with overall survival [32].”
    Quote in the paperSubstantial claim0 of 1 sources checked
    • cites [32] Gao Y, Liu Y, Sun L, Ouyang X, Zhu C, Qin X. MAD2L1 functions as a novel diagnostic and predictive biomarker in cholangiocarcinoma. Genet Test Mol Biomarkers. 2021;25:685–95. source

  • Probably not a substantial claim: may not pass S3 (load-bearing).

    Premisecitation

    c-Myc is the critical effector of Akt/FBXW7ΔF-dependent cholangiocarcinogenesis.

    “Interestingly, c-Myc is the critical effector of Akt/FBXW7ΔF-dependent cholangiocarcinogenesis [31].”
    Quote in the paperNot a substantial claim0 of 1 sources checked
    • cites [31] Fan W, Huang X, Chen C, Gray J, Huang T. TBX3 and its isoform TBX3+2a are functionally distinctive in inhibition of senescence and are overexpressed in a subset of breast cancer cell lines. Cancer Res. 2004;64:5132–9. source

  • Probably not a substantial claim: may not pass S3 (load-bearing).

    Premisecitation

    Cases of liver cirrhosis evolving into osteosarcomas have been previously described.

    “and cases of liver cirrhosis evolving into osteosarcomas have been previously described [31].”
    Quote in the paperNot a substantial claim0 of 1 sources checked
    • cites [31] Fan W, Huang X, Chen C, Gray J, Huang T. TBX3 and its isoform TBX3+2a are functionally distinctive in inhibition of senescence and are overexpressed in a subset of breast cancer cell lines. Cancer Res. 2004;64:5132–9. source

  • Probably not a substantial claim: may not pass S3 (load-bearing). May combine two claims.

    Interpretationcitation

    The diverse behavior of TBX3 may be due to the expression of alternatively spliced isoforms and the cell/tissue context.

    “This diverse behavior may be due to the expression of alternatively spliced isoforms [31] and the cell/tissue context.”
    Quote in the paperNot a substantial claim0 of 1 sources checked
    • cites [31] Fan W, Huang X, Chen C, Gray J, Huang T. TBX3 and its isoform TBX3+2a are functionally distinctive in inhibition of senescence and are overexpressed in a subset of breast cancer cell lines. Cancer Res. 2004;64:5132–9. source

  • Probably not a substantial claim: may not pass S2 (checkable), S3 (load-bearing), S4 (based).

    Interpretationnone given

    Presumably, only specific degrees of expression of TBX3 and interactors result in mesenchymal commitment and differentiation.

    “Presumably, only specific degrees of expression of TBX3 and interactors result in mesenchymal commitment and differentiation.”
    Quote in the paperNot a substantial claim

  • Probably not a substantial claim: may not pass S2 (checkable), S3 (load-bearing).

    Interpretationreasoning

    The interaction with tissue- or tumor-specific transcription factors might dictate the net effect of TBX3 in a particular tumor type.

    “In this regard, the interaction with tissue- or tumor-specific transcription factors might dictate the net effect of TBX3 in a particular tumor type.”
    Quote in the paperNot a substantial claim

  • Probably not a substantial claim: may not pass S3 (load-bearing).

    Interpretationreasoning

    The TBX3/c-Myc and/or Akt cooperation may also be responsible for osteosarcoma-like lesions occurrence in Akt/FBXW7ΔF/TBX3 mice.

    “Thus, the TBX3/c-Myc and/or Akt cooperation may also be responsible for osteosarcoma-like lesions occurrence in Akt/FBXW7ΔF/TBX3 mice.”
    Quote in the paperNot a substantial claim

  • Probably not a substantial claim: may not pass S3 (load-bearing).

    Premisecitation

    Synergistic activity of TBX3, c-Myc, and AKT has been revealed in rhabdomyosarcoma.

    “In addition, synergistic activity of TBX3, c-Myc, and AKT has been revealed in rhabdomyosarcoma [31].”
    Quote in the paperNot a substantial claim0 of 1 sources checked
    • cites [31] Fan W, Huang X, Chen C, Gray J, Huang T. TBX3 and its isoform TBX3+2a are functionally distinctive in inhibition of senescence and are overexpressed in a subset of breast cancer cell lines. Cancer Res. 2004;64:5132–9. source

  • Probably not a substantial claim: may not pass S3 (load-bearing).

    Premisecitation

    Human hepatoblastomas with an osteoid component are well-known in clinical practice.

    “Intriguingly, human hepatoblastomas with an osteoid component are well-known in clinical practice [31]”
    Quote in the paperNot a substantial claim0 of 1 sources checked
    • cites [31] Fan W, Huang X, Chen C, Gray J, Huang T. TBX3 and its isoform TBX3+2a are functionally distinctive in inhibition of senescence and are overexpressed in a subset of breast cancer cell lines. Cancer Res. 2004;64:5132–9. source

  • Probably not a substantial claim: may not pass S3 (load-bearing).

    Premisecitation

    TBX3 cooperates with c-Myc to induce the transformation of sarcoma-initiating cells.

    “Previously, it has been shown that TBX3 cooperates with c-Myc to induce the transformation of sarcoma-initiating cells [31].”
    Quote in the paperNot a substantial claim0 of 1 sources checked
    • cites [31] Fan W, Huang X, Chen C, Gray J, Huang T. TBX3 and its isoform TBX3+2a are functionally distinctive in inhibition of senescence and are overexpressed in a subset of breast cancer cell lines. Cancer Res. 2004;64:5132–9. source

  • Probably not a substantial claim: may not pass S3 (load-bearing).

    Premisecitation

    The protein product of MAD2L1 is part of the mitotic spindle assembly checkpoint that halts anaphase until proper chromosomal alignment occurs.

    “The protein product of MAD2L1 is part of the mitotic spindle assembly checkpoint that halts anaphase until proper chromosomal alignment occurs [32].”
    Quote in the paperNot a substantial claim0 of 1 sources checked
    • cites [32] Gao Y, Liu Y, Sun L, Ouyang X, Zhu C, Qin X. MAD2L1 functions as a novel diagnostic and predictive biomarker in cholangiocarcinoma. Genet Test Mol Biomarkers. 2021;25:685–95. source
31 of 31

  • The paper relies on this without support.

    TBX3 function in intrahepatic cholangiocarcinoma is undetermined.

    “However, TBX3 function remains undetermined in intrahepatic cholangiocarcinoma (iCCA), a deadly primary liver malignancy with few systemic treatment options.”
    “Given the ongoing debate regarding the role of TBX3 in liver cancer and the limited knowledge of its specific involvement in iCCA”

  • The paper relies on this without support.

    Injection of pCMV-Cre into Tbx3flox/flox mice deletes Tbx3 in liver cells, allowing expression of Akt/FBXW7ΔF in Tbx3-null cells.

    “This strategy allows the expression of Akt/FBXW7ΔF into Tbx3-null liver cells.”

  • The paper relies on this without support.

    Systemic treatment options for iCCA have limited success partially due to incomplete understanding of iCCA pathogenesis and biology.

    “Overall, systemic treatment options have limited success partially due to the incomplete understanding of iCCA pathogenesis and biology.”
3 of 3

Self-check

Percentages, sums and reported differences re-computed from the paper’s own numbers.

Each contradiction between two passages listed here was confirmed by an independent check.

  • Predicted TBX3 binding motif is assigned to two promoters

    Minor

    The Results place the candidate binding motif in the TBX3 promoter, but describe the subsequent binding test and conclusion as concerning the MAD2L1 promoter.

  • Mouse euthanasia threshold changes from liver weight to tumor weight

    Minor

    The Methods define the threshold using liver weight, but the Fig. 2 caption describes it as tumor weight.

  • Fig. 4 identifies the wrong control group

    Minor

    The study design identifies Akt/FBXW7ΔF/pCMV as the control, while the Fig. 4C caption calls the pictured control Akt/FBXW7ΔF/pT3.

3 of 3

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Authors
Deng, S., Lu, X., Wang, X., Liang, B., Xu, H., Yang, D., Cui, G., Yonemura, A., Paine, H., Zhou, Y., Zhang, Y., Simile, M. M., Urigo, F., Evert, M., Calvisi, D. F., Green, B. L., & Chen, X.
Affiliations
  • University of California, San Francisco (US)
  • Sun Yat-sen University (CN)
  • University of Hawaii Cancer Center
  • Chongqing University of Technology (CN)
  • University of Sassari (IT)
  • University of Regensburg (DE)
Corresponding author
  • Green, Benjamin L.
  • Chen, Xin
ORCID
Venue
Cell Death & Disease
Publisher
Springer Science and Business Media LLC
Published
2024-06-22
Volume
15
Issue
6
Pages
441-441
ISSN
2041-4889 (ISSN-L)
DOI
10.1038/s41419-024-06839-8
PMC
PMC11193761
OpenAlex
W4399916896
Semantic Scholar
aaf0c875971d079bd0cac31dd4af33549d7ed6fb
Cited by
2
References
34
Citations by year
2024: 2
FWCI
0.76
Type
journal article
Language
en
Open access
Licence
Locations
  • Cell Death and Disease (publishedVersion, cc-by)
  • PubMed (publishedVersion)
  • eScholarship (California Digital Library) (submittedVersion, cc-by)
  • PubMed Central (submittedVersion, cc-by)
  • DOAJ (DOAJ: Directory of Open Access Journals) (submittedVersion)
  • UnissResearch (Università degli Studi di Sassari) (submittedVersion)
Funders
  • Foundation for the National Institutes of Health (R01CA228483, R01CA239251, R01CA250227)
  • UC | UC San Francisco | Claude D. Pepper Older Americans Independence Center, University of California San Francisco (P30DK026743)
  • National Natural Science Foundation of China (82202981)
  • Cholangiocarcinoma Foundation
Topics
  • Health Sciences › Medicine › Surgery › Cholangiocarcinoma and Gallbladder Cancer Studies
  • Health Sciences › Medicine › Surgery › Pediatric Hepatobiliary Diseases and Treatments
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Cell Biology › Hippo pathway signaling and YAP/TAZ
Keywords
TBX3, intrahepatic cholangiocarcinoma, tumor suppressor, Mad2, cholangiocarcinogenesis, gene expression regulation, tumor growth inhibition, patient survival, transcriptional repression
MeSH terms
Animals, Bile Duct Neoplasms, Humans, Gene Expression Regulation, Neoplastic, Cholangiocarcinoma, T-Box Domain Proteins, Cell Line, Tumor, Cell Proliferation, Mice, Carcinogenesis

About these checks

Registry, retraction, Cites and statistics results are computed, not judged, with one exception: a printed reference that no index lists is read once by a model before it counts as a citation error. Claims are extracted by a model and kept only when their quote appears verbatim in the text; each cited source is then checked, with a model’s help, for whether it says what the claim relies on. Those verdicts can be wrong — read the claim’s quote and the cited source before relying on one.

Claims read by deepseek-v4.1-flash under definition version 3 (extract-v3) on 2026-10-22.

Classified by jev-1.13-20260917 (gold-guideline-v0.2) on 2026-10-22.

Cited sources are checked by gpt-6-sol.

Statistics and arithmetic are recomputed, not judged.

Confidence bands are shown only for fields calibrated on our annotated test set. None is calibrated yet, so no band is shown.

What counts as a claim ›How we measure confidence ›How we check this ›

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