bims-merabr 2026-06-14 papers

bims-merabr

Biomed News

on Metabolic rewiring in aggressive breast cancer

Issue of 2026–06–14
eleven papers selected by
Barbara Mensah Sankofi, University of Oklahoma Health Sciences Center

Breast Cancer and Metabolic Dysfunction-Associated Steatotic Liver Disease.
PDZK1 disassembles HER2-HSP90 complexes to promote ubiquitin-mediated HER2 degradation and overcome therapy resistance.
Bergenin Suppresses Glycolysis and Malignant Progression in Breast Cancer via the IGF1R-MAPK Signaling Pathway.
The HDAC Inhibitor RM-3-22 Suppresses Triple-Negative Breast Cancer by Uncoupling Autophagy to Drive FSP1-Dependent Ferroptosis.
Targeting neuropilin-1 attenuates 4-hydroxytamoxifen-induced stemness and sensitizes ERα-re-expressing TNBC to tamoxifen.
Epirubicin-Induced Upregulation of miR-143-3p and miR-145 Enhances Cytotoxicity in MCF-7, MDA-MB-231, and BT-474 Breast Cancer Cell Lines.
A peptide-induced regression of low-grade malignant breast tumor via activation of autophagy in breast cancer cells.
Adiponectin Preserves Follicles through ADIPOR1/ADIPOR2-Driven Fatty Acid Metabolism.
Genome Organizer SATB1 selectively activates a defined subset of EMT genes driving metastatic breast cancer.
NSUN2/m5C/TPI1 axis promotes glycolysis and immune evasion in breast cancer.
GLIS3 drives epithelial-mesenchymal transition and cancer stem-like traits in stomach adenocarcinoma via TGFBR3-Hedgehog signaling.

Int J Mol Sci. 2026 May 29. pii: 4919. [Epub ahead of print]27(11):

Breast Cancer and Metabolic Dysfunction-Associated Steatotic Liver Disease.

Damaris G Nieva-Ramírez, David Luna-Pérez, Misael Uribe, Natalia Nuño-Lámbarri.

  Breast cancer remains the most frequently diagnosed malignancy among women worldwide, while metabolic dysfunction-associated steatotic liver disease (MASLD) represents the leading cause of chronic liver disease, reflecting a global burden of metabolic dysfunction. Increasing evidence suggests that MASLD is associated with breast cancer development and progression; however, whether this relationship reflects an independent effect of hepatic metabolic dysfunction or the broader metabolic environment remains uncertain. This review synthesizes current epidemiological, clinical, and mechanistic data linking hepatic metabolic dysfunction to breast carcinogenesis. Population-based studies consistently demonstrate an association between hepatic steatosis and increased breast cancer incidence, particularly in postmenopausal and metabolically vulnerable populations, as well as poorer oncological outcomes. Mechanistically, MASLD promotes a systemic pro-tumorigenic environment through interconnected pathways, including insulin resistance, hormonal dysregulation with increased estrogen bioavailability, chronic inflammation, oxidative stress, lipid metabolic reprogramming, and gut-liver axis disruption. Hepatokines, particularly fibroblast growth factor 21 (FGF21), emerge as key mediators of tumor progression and potential biomarkers of metabolic vulnerability, while Fetuin-A and angiopoietin-like protein 8 (ANGPTL8) further support the liver's endocrine role in oncogenic signaling. Preclinical evidence highlights fatty acid oxidation as a metabolic dependency in aggressive breast cancer subtypes, suggesting novel therapeutic targets. Despite consistent associations, causality remains unproven. Future prospective studies are needed to determine whether targeting metabolic dysfunction can improve breast cancer prevention and outcomes.

Keywords:  breast neoplasms; hepatokines; inflammation; lipid metabolism; liver

DOI:  https://doi.org/10.3390/ijms27114919
J Adv Res. 2026 Jun 06. pii: S2090-1232(26)00444-3. [Epub ahead of print]

PDZK1 disassembles HER2-HSP90 complexes to promote ubiquitin-mediated HER2 degradation and overcome therapy resistance.

Xuedi Cao, Chen Qin, Yu Guo, Yumeng Yang, Qiong Qin, Jing Li, Yang Yang, Xuan Qi, Haibo Wang, Lijie Zhang, Tianzhang Hou, Xiaomei Yang, Ran Song, Duiping Feng, Hongyan Jia, Junqi He.

   INTRODUCTION: Limited therapeutic response and disease recurrence remain major challenges in HER2-positive breast cancer, largely driven by sustained HER2 stabilization and persistent oncogenic signaling mediated by HSP90. Therefore, targeting HER2 protein stability may represent a promising strategy to improve therapeutic responsiveness and suppress HER2-driven tumor progression.
OBJECTIVES: This study investigated the mechanisms by which PDZK1 regulates HER2 stability and evaluated its clinical and therapeutic relevance in HER2-positive breast cancer.
METHODS: Integrated transcriptomic analyses identified PDZK1 as a potential tumor suppressor during HER2-positive breast cancer progression and recurrence. Functional studies using 2D/3D cell assays, xenograft and Pdzk1-deficient mouse models, tissue microarrays, and immunohistochemistry were performed to evaluate PDZK1 function. Molecular docking, co-immunoprecipitation, GST pull-down, and ubiquitination assays were used to characterize PDZK1/HER2/HSP90 interactions. RNA-seq, PAM50 classification, tamoxifen-resistant models, and synergy analyses were additionally conducted to investigate the relationship between PDZK1 deficiency and endocrine resistance.
RESULTS: PDZK1 expression was significantly reduced in HER2-positive breast cancer and was associated with poor prognosis and unfavorable chemotherapy response. Mechanistically, PDZK1 directly bound to HER2, disrupted the HER2/HSP90 complex, and recruited the HSP70/CHIP complex to promote HER2 polyubiquitination and degradation. PDZK1 loss also promoted tamoxifen resistance in ER-positive breast cancer models. Furthermore, pharmacological induction of PDZK1 by fenofibrate enhanced the efficacy of combinatorial therapeutic regimens in resistant breast cancer models. These findings suggest PDZK1 may serve as a potential biomarker associated with chemotherapy response and endocrine resistance.
CONCLUSIONS: PDZK1 functions as a regulator of HER2 stability and may serve as a potential biomarker and therapeutic target in HER2-positive breast cancer. Pharmacological upregulation of PDZK1 may represent a promising combinatorial therapeutic strategy for overcoming therapeutic resistance in breast cancer.

Keywords:  Chemotherapy response; Endocrine resistance; HER2 stability; HER2-positive breast cancer; PDZK1

DOI:  https://doi.org/10.1016/j.jare.2026.05.048
Ann Clin Lab Sci. 2026 Mar;56(2): 214-226

Bergenin Suppresses Glycolysis and Malignant Progression in Breast Cancer via the IGF1R-MAPK Signaling Pathway.

Minhui Fan, Qingyu Li, Renya Shuai, Yunfei Shen, Fanglin Zhang.

OBJECTIVE: This study endeavors to unravel the effects and mechanisms of bergenin (Ber) in suppressing breast cancer (BC) progression through modulating the IGF1R-MAPK signaling pathway, using an integrative approach combining network pharmacology and bioinformatics.
METHODS: Ber-BC targets and related signaling pathways were systematically identified via network pharmacology and bioinformatics analyses. Key targets were further prioritized via machine learning algorithms, and their binding affinities with Ber were evaluated through molecular docking. IGF1R-overexpressing BC cell lines were subsequently established and pretreated with the MAPK inhibitor SB203580 for one hour before Ber administration. Functional assays were conducted to assess cellular proliferation, migration, glucose uptake, as well as intracellular pyruvate, lactate, and ATP. The extracellular acidification rate was derived via a Seahorse XF Analyzer. IGF1R expression was visualized via immunofluorescence. A murine xenograft model was generated through subcutaneous BC cell injection. Protein levels of HK2, PKM2, IGF1R, and phosphorylated p38 MAPK were quantified via western blotting, while immunohistochemistry was performed to evaluate Ki67 and IGF1R expression in tumor tissues.
RESULTS: Network pharmacology, bioinformatics, and MD identified eight pivotal targets (KDM5B, IGF1R, ADA, LGALS9, EGFR, SERPINE2, PHGDH, and ALOX5), among which IGF1R was prominently linked to the MAPK signaling pathway. In vitro, Ber markedly suppressed BC cell proliferation, migration, as well as glycolytic activity, concomitant with reduced IGF1R expression and diminished p38 MAPK phosphorylation. Rescue experiments proved that IGF1R overexpression notably ameliorated proliferation, migration, and glycolysis in Ber-treated BC cells, whereas SB203580 administration effectively reversed these effects. In vivo, Ber treatment significantly suppressed tumor growth and downregulated HK2, PKM2, and IGF1R protein expression, as well as p38 MAPK phosphorylation.
CONCLUSION: Ber suppresses glycolysis and malignant progression in BC through modulation of the IGF1R-MAPK signaling pathway.

Keywords: BC; Ber; Glycolysis; IGF1R; MAPK signaling pathway
Transl Res. 2026 Jun 12. pii: S1931-5244(26)00121-0. [Epub ahead of print]

The HDAC Inhibitor RM-3-22 Suppresses Triple-Negative Breast Cancer by Uncoupling Autophagy to Drive FSP1-Dependent Ferroptosis.

Essha Chatterjee, Biswajit Dey, Santanu Basak, Ram Sharma, Anamika Sharma, Hoshiyar Singh, Vigneshwar Reddy Ashireddygari, Prasad Tammineni, Kunal Nepali, Santosh Kumar Guru.

  Triple negative breast cancer (TNBC) remains one of the difficult subtypes to treat since it holds aggressiveness, a high recurrence rate, and is devoid of targeted therapy options. Taking these challenges into view, we developed RM-3-22, a hydroxamic acid-based histone deacetylase (HDAC) inhibitor, as a new therapeutic entity, particularly to target TNBC. Various experiments were performed using TNBC cell lines and a xenograft mouse model. Results revealed that RM-3-22 exhibited greater cytotoxicity than SAHA, an FDA-approved inhibitor with IC50 values of 6.49, 7.3, and 3.89 μM against MDA-MB-231, BT-549, and 4T1 Cells, respectively. RM-3-22 exhibited a strong anti-metastatic profile, as evidenced by impaired migratory capabilities of TNBC cells and significant disruption of the architecture of TNBC cell 3D spheroids. Furthermore, RM-3-22 suppressed TNBC progression by targeting autophagy and ferroptosis, as evidenced by reduced p62 and FSP1 expression and increased LC3B and ACSL4 expression. Our research explored the key axis where induction of autophagy is required for RM-3-22-mediated FSP1-dependent ferroptosis in the TNBC model. Surprisingly, as far as we know, this study is the first to show that HDAC inhibitors could induce autophagy-dependent ferroptosis as a cell death mechanism to halt TNBC progression. These results were further validated by the effective reduction in tumour growth in the xenograft mouse model. Collectively, RM-3-22 could be a promising alternative inhibitor for TNBC by targeting the autophagy-ferroptosis mechanism(s).

Keywords:  Autophagy; Ferroptosis; HDAC inhibitor; Novel anticancer molecule; TNBC

DOI:  https://doi.org/10.1016/j.trsl.2026.06.010
Biochem Pharmacol. 2026 Jun 12. pii: S0006-2952(26)00500-9. [Epub ahead of print] 118163

Targeting neuropilin-1 attenuates 4-hydroxytamoxifen-induced stemness and sensitizes ERα-re-expressing TNBC to tamoxifen.

Qinglong Yu, Wei Tang, Yani Xiao, Ying Kong, Jinguang Liu, Tongfang Li, Xiangping Li, Li Zhang, Rong Rong.

  Reactivating functional estrogen receptor alpha (ERα) expression represents a promising strategy for tamoxifen (TAM)-based endocrine therapy in triple-negative breast cancer (TNBC). However, TAM and its metabolites may exert potential oncogenic effects, which could compromise TAM efficacy in TNBC. Here, we report that low-dose 4-hydroxytamoxifen (4-OHT), an active metabolite of TAM, enhances stemness in TNBC cells via the Neuropilin-1 (NRP-1)/Calpain-2 (CAPN2)/β-catenin axis. Mechanistically, 4-OHT stimulates CAPN2 activation by promoting its membrane localization, increasing substrate cleavage, and inducing degradation of its endogenous inhibitor calpastatin. Activated CAPN2 suppresses ubiquitin-mediated degradation of β-catenin, leading to elevated β-catenin stability and intracellular accumulation. This enhances β-catenin transcriptional activity and ultimately promotes stemness in TNBC cells. Kaplan-Meier survival analysis showed that high NRP1 expression correlates with poorer overall survival in breast cancer patients undergoing endocrine therapy. Moreover, 4-OHT triggers Ca2+ release and ERK phosphorylation in TNBC cells through the NRP-1/PLC-γ1 pathway, thereby activating CAPN2. Finally, we demonstrate that the NRP-1 inhibitor EG00229 sensitizes ERα-re-expressing TNBC cells to TAM treatment. Our study uncovers a molecular mechanism by which 4-OHT promotes TNBC stemness via the NRP-1/CAPN2/β-catenin signaling cascade. Targeting NRP-1 may serve as a valuable strategy to improve TAM efficacy in ERα-re-expressing TNBC.

Keywords:  4-hydroxy-tamoxifen; Calpain-2; Neuropilin-1; Stemness; Tamoxifen; Triple-negative breast cancer; β-catenin

DOI:  https://doi.org/10.1016/j.bcp.2026.118163
Biochem Genet. 2026 Jun 08.

Epirubicin-Induced Upregulation of miR-143-3p and miR-145 Enhances Cytotoxicity in MCF-7, MDA-MB-231, and BT-474 Breast Cancer Cell Lines.

Morteza Rajabi, Niki Ghambari Mohammadi, Mahsa Mombeyni, Roudabeh Behzadi Andouhjerdi.

  Drug resistance poses a significant challenge to effective breast cancer chemotherapy. This study investigated the regulatory effects of Epirubicin on miR-143-3p and miR-145 expression in BT-474, MCF-7, and MDA-MB-231 breast cancer cell lines. These miRNAs, known as tumor suppressors, are often downregulated in breast carcinoma, correlating with poorer prognosis. Cells were treated with Epirubicin, and miR-143-3p and miR-145 expression was quantified via qRT-PCR. Epirubicin IC50 values were 0.42 µg/mL (MCF-7), 2.94 µg/mL (MDA-MB-231), and 0.86 µg/mL (BT-474) after 48 h, demonstrating dose- and time-dependent cytotoxicity. Treatment significantly upregulated miR-143-3p and miR-145 expression, with MCF7 cells showing a 3.5-fold increase in miR-145 (p < 0.01) and a 2.8-fold increase in miR-143-3p (p < 0.05) compared to controls. Cell viability decreased by up to 70% at 6 µg/mL Epirubicin in MCF-7 cells, with apoptosis rates rising to 8.65% (MCF-7) and 9.75% (MDA-MB-231) after 24 h (p < 0.01). Colony formation was reduced by 60-80% across cell lines (p < 0.001), and wound closure was inhibited by 40-50% in MDA-MB-231 cells at 24 h (p < 0.0001). These findings suggest that Epirubicin enhances therapeutic efficacy by upregulating miR-143-3p and miR-145, potentially mitigating drug resistance. This highlights their potential as therapeutic targets for improving outcomes in resistant breast cancer subtypes.

Keywords:  Apoptosis; Breast neoplasms; Epirubicin; MiR-143-3p; MiR-145; MicroRNAs

DOI:  https://doi.org/10.1007/s10528-026-11408-z
Mol Med. 2026 Jun 09.

A peptide-induced regression of low-grade malignant breast tumor via activation of autophagy in breast cancer cells.

Saibal Saha, Rishikesh Kumar, Snehanjan Sarangi, Animesh Gope, Ananda Pal, Rima Tapader, Niraj Nag, Sushmita Bhattacharya, Amit Pal.

  Peptides have recently gained much attention for their therapeutic role in several diseases without having significant side effects. These are short chain amino acids and can be synthesized chemically with a great extent of purity, which renders them very effective, precise and safe for the body. So far, very few artificially synthesized peptides have been developed that have autophagy-inducing properties. Among them, a very small number of peptides have been demonstrated to have anti-tumorigenic effect by modulating autophagy with most of them lacking specificity for target cells. This study aims to investigate the regulation of autophagy through noncanonical activation of protease-activated receptor 1 (PAR1) by hemagglutinin protease (HAP). Unlike the canonical activation of PAR1 by thrombin, which enhances cell proliferation via mechanistic target of rapamycin (mTOR) signaling, HAP-induced noncanonical activation downregulates mTOR activation and triggers autophagy in both estrogen receptor positive/ progesterone receptor positive/ HER2 negative (ER+/PR+/HER2-) and triple negative (ER-/PR-/HER2-) breast cancer cells. This noncanonical activation generates a N-terminal sequence in PAR1. When this sequence is mimicked by a synthetic peptide, it induces autophagy independently of HAP in the breast cancer cells in vitro. Further in vivo investigation in BALB/c mouse model of low-grade malignant breast tumor reveals that the peptide-induced autophagy significantly inhibits tumor growth and delays tumor progression. Importantly, the lack of PAR1 expression in normal, healthy breast epithelial cells facilitates the peptide to selectively target breast cancer cells with relatively high PAR1 expression, highlighting its potential as a therapeutic agent against low-grade malignant breast tumor. These findings provide valuable insights into autophagy activation by a synthetic peptide that targets breast cancer cells with high PAR1 expression and for the first time shows peptide mediated targeted therapy of low-grade malignant breast tumor.

Keywords:  Hemagglutinin protease; Peptide; Peptide-induced autophagy; Protease-activated receptor 1

DOI:  https://doi.org/10.1186/s10020-026-01514-4
Mol Metab. 2026 Jun 10. pii: S2212-8778(26)00076-1. [Epub ahead of print] 102392

Adiponectin Preserves Follicles through ADIPOR1/ADIPOR2-Driven Fatty Acid Metabolism.

Dae Hyun Lee, Jae Young Shin, Hyeri Park, Jin Seok, Kyu Hwan Na, Young-Ran Kim, Gi Jin Kim.

   BACKGROUND: Adiponectin is a key regulator of glucose and lipid metabolism that improves insulin sensitivity and promotes mitochondrial fatty acid oxidation via ADIPOR1 and ADIPOR2. Ovarian lipid accumulation contributes to metabolic reproductive disorders such as polycystic ovary syndrome (PCOS), yet current hormone-based therapies have limited efficacy and potential adverse effects. We evaluated whether placenta-derived mesenchymal stem cells (PDMSCs) mitigate ovarian lipotoxicity by restoring adiponectin signaling.
METHODS: A thioacetamide (TAA) induced rat model of metabolic dysfunction with ovarian lipotoxicity was treated by intravenous transplantation of PDMSCs (2 × 106) cells. Hepatic and ovarian phenotypes were assessed four weeks after transplantation. In parallel, PDMSCs were cocultured with TAA-treated granulosa and primary theca cells, with or without siRNA-mediated knockdown of ADIPOR1 and/or ADIPOR2.
RESULTS: PDMSCs transplantation improved systemic insulin resistance and dyslipidemia and partially restored hepatic and ovarian architecture. PDMSCs treatment increased circulating and ovarian ADIPONECTIN levels and upregulated Adipor1/2 in ovarian tissue, accompanied by activation of the ADIPOR1/2 / Fatty acid driven axis and enhanced mitochondrial fatty acid oxidation. These changes were associated with reduced ovarian lipid accumulation and improved endocrine homeostasis, including normalization of anti-Müllerian hormone (AMH), estradiol, and androgen levels, preservation of the primordial follicle pool, and induction of BMP15 expression. In vitro ADIPOR1/2 silencing abrogated these protective effects, supporting a requirement for adiponectin receptor signaling.
CONCLUSION: PDMSCs ameliorate systemic and ovarian metabolic dysfunction in a TAA-induced model, consistent with adiponectin, ADIPOR1/2 dependent mitochondrial metabolic reprogramming. PDMSCs restore both metabolic and reproductive competence in the context of hepatic-ovarian metabolic crosstalk. These findings support PDMSC-based therapy as a mechanistically informed, multi-target strategy for the treatment of PCOS and metabolic-associated ovarian dysfunction.

Keywords:  Adiponectin; Fatty acid oxidation; Mesenchymal stem cell; Metabolic syndrome

DOI:  https://doi.org/10.1016/j.molmet.2026.102392
bioRxiv. 2026 Jun 02. pii: 2026.05.29.728584. [Epub ahead of print]

Genome Organizer SATB1 selectively activates a defined subset of EMT genes driving metastatic breast cancer.

Yoshinori Kohwi, Yelena Vayn, Mari Grange, Bao Ho, Diane Heditsian, Terumi Kohwi-Shigematsu.

SATB1 reshapes chromatin architecture and transcriptional programs to promote breast cancer metastasis. However, its key downstream effectors remain incompletely defined. Here, we aimed to identify actionable drivers of invasion by focusing on epithelial-mesenchymal transition (EMT) genes. We identified 98 of 300 curated EMT-promoting genes as direct SATB1 targets in human breast epithelial cells (MCF10A-1) rendered tumorigenic with metastatic traits by SATB1 transduction, using Global Run-On Sequencing (GRO-seq) to measure nascent transcripts. These SATB1-activated EMT genes regulate extracellular matrix remodeling, hypoxia-responsive transcriptional programs, and tumor microenvironmental programs linking angiogenesis and immune evasion, collectively enhancing metastatic competence. Triple-negative breast cancer (TNBC) is a heterogeneous disease characterized by frequent metastasis and chemoresistance. Among the four TNBC molecular subtypes, the 98 SATB1-regulated EMT genes were significantly enriched and activated in the Basal-like 2 (BL2) subtype (Fisher's exact test: p = 4.53e-9), which is associated with aggressive behavior, poorer clinical outcomes, and reduced treatment responsiveness. In contrast, SATB1-independent EMT genes showed no enrichment in BL2, indicating selective regulation of EMT genes by SATB1. We further analyzed nascent transcripts induced by the environmental carcinogen benzo[a]pyrene (B[a]P), a known breast carcinogen. Half of the 72 EMT genes activated after short-term B[a]P exposure overlapped with SATB1-dependent EMT genes, indicating that two distinct etiologies, SATB1 and B[a]P, converge on a largely shared network of invasion-promoting genes. These results show that EMT genes are not globally or randomly activated in breast cancer but are selectively activated, defining an EMT gene network associated with metastatic risk. This gene signature may serve as a prognostic marker pending further validation.

DOI: https://doi.org/10.64898/2026.05.29.728584
J Transl Med. 2026 Jun 06.

NSUN2/m5C/TPI1 axis promotes glycolysis and immune evasion in breast cancer.

Xinming Song, Qihang Li, Ziteng Xiao, Haiqing Luo, Xiangyong Li.

   BACKGROUND: Aerobic glycolysis and immune evasion drive tumor development. 5-methylcytosine (m5C) methyltransferase NSUN2 acts as an oncogene in breast cancer (BC), but the molecular mechanisms remain incompletely elucidated. This study aimed to investigate the role of NSUN2 in BC and the underlying mechanisms.
METHODS: In vitro experiments were conducted to determine cell proliferation, glycolysis, and immune evasion. Tumor growth was evaluated using a tumor-bearing mouse model. The interplay among NSUN2, YBX1, and TPI1 was evaluated by RNA immunoprecipitation, methylated RNA immunoprecipitation, and dual-luciferase reporter assay.
RESULTS: The results showed that NSUN2 and TPI1 were highly expressed in BC cells. NSUN2 knockdown inhibited BC cell proliferation, glycolysis, and immune evasion, which was reversed by TPI1 overexpression. NSUN2 enhanced TPI1 stability by facilitating the m5C modification, which was recognized by YBX1. Moreover, overexpression of TPI1 reversed the inhibition of tumor growth, glycolysis, and immune cell infiltration induced by NSUN2 silencing in vivo.
CONCLUSIONS: In conclusion, NSUN2 accelerates the progression of BC by promoting glycolysis and immune evasion. Mechanistically, NSUN2 promotes m5C methylation of TPI1 in a YBX1-dependent manner, which is associated with increased PD-L1 expression and immune-evasive phenotypes. These findings suggest that NSUN2 may be an effective therapeutic target for BC.

Keywords:  Breast cancer; Glycolysis; Immune evasion; M5C methylation; NSUN2; PD-L1; TPI1

DOI:  https://doi.org/10.1186/s12967-026-08390-w
Front Oncol. 2026 ;16 1826297

GLIS3 drives epithelial-mesenchymal transition and cancer stem-like traits in stomach adenocarcinoma via TGFBR3-Hedgehog signaling.

Qi Xiao, Hongyang Deng, Jipin Li, Yijun Zheng, Youcheng Zhang.

   Background: GLIS family zinc finger 3 (GLIS3) is a transcription factor implicated in multiple malignancies, but its role in stomach adenocarcinoma (STAD) and its downstream effector axis remain unclear. We investigated whether GLIS3 coordinates epithelial-mesenchymal transition (EMT) and cancer stem cell (CSC)-like programs in STAD through a signaling cascade.
Methods: GLIS3 expression and prognostic associations were analyzed using transcriptomic datasets from The Cancer Genome Atlas, Genotype-Tissue Expression, and Gene Expression Omnibus. GLIS3 protein levels were evaluated by immunohistochemistry in 133 paired STAD tissues. Survival was assessed by Kaplan-Meier analysis together with multivariable regression models. Gain- and loss-of-function studies were performed in AGS and HGC-27 cells to evaluate proliferation, migration, EMT and CSC-like phenotypes. Mechanistic experiments interrogated a GLIS3-transforming growth factor beta receptor 3 (TGFBR3)-Hedgehog axis using ChIP-qPCR, dual-luciferase assays, co-immunoprecipitation, GLI reporter assays, and vismodegib treatment. Tumor growth was assessed in xenograft models.
Results: GLIS3 was consistently upregulated in STAD across public datasets and in the clinical cohort (median H-score: 112 vs 39, P < 0.001) and was associated with adverse clinicopathological features and poor survival. In the TCGA cohort, higher GLIS3 expression was associated with greater nodal burden, more advanced nodal and metastatic status, and worse vital status. In multivariable Cox models restricted to cases with defined stage classifications, GLIS3 remained independently associated with worse overall survival in the primary model (hazard ratio = 1.45, 95% confidence interval: 1.04-2.02; P = 0.031), with a consistent result in the sensitivity model. Functionally, GLIS3 enhanced proliferation and migration and promoted EMT marker switching and CSC-like traits. Mechanistically, GLIS3 transcriptionally activated TGFBR3 and increased Hedgehog pathway activity; bidirectional rescue experiments showed that TGFBR3 overexpression partially restored, whereas TGFBR3 silencing attenuated, GLIS3-driven EMT/CSC-like phenotypes and growth-related functions. Xenograft experiments supported an in vivo growth-promoting role of the GLIS3-TGFBR3-Hedgehog axis.
Conclusions: GLIS3 identifies an aggressive STAD phenotype and engages a TGFBR3-Hedgehog program linked to EMT and CSC-like features. These findings support further evaluation of GLIS3 as a biomarker for risk stratification and provide a rationale for biomarker-guided targeting of the GLIS3-TGFBR3-Hedgehog axis in STAD.

Keywords:  CSC; EMT; GLIS3; Hedgehog; STAD

DOI:  https://doi.org/10.3389/fonc.2026.1826297