TRIM47 promotes cell viability, cell cycle progression, metabolic reprogramming, and inhibits apoptosis of glioma via the NF-κB signaling pathway

Introduction

Glioma represents the majority of malignant brain tumors, with glioblastoma (GBM) being the most aggressive subtype, distinguished by fast proliferation, invasive properties, and resistance to conventional treatment strategies (1,2). Even with improvements in surgical techniques, radiotherapy, and chemotherapy, the prognosis remains poor, with a median survival of 12–15 months (3). Therefore, identifying key oncogenic drivers and unraveling their molecular mechanisms are crucial for developing novel therapeutic strategies.

Metabolic reprogramming, including the move towards aerobic glycolysis, is a distinctive feature of cancer cells, supplying the energy and biosynthetic materials necessary for their swift proliferation (4). Microglia display unique phenotypic forms at various phases of central nervous system disorders, and these polarization processes are intricately linked with changes in glucose metabolism (5). During neuroinflammation, activated microglia and astrocytes opt for aerobic glycolysis rather than oxidative phosphorylation (6). PKM2 is a significant enzyme in glycolysis that catalyzes the last phase of glycolysis, resulting in the creation of pyruvate and adenosine triphosphate (7-9). The nuclear factor kappa B (NF-κB) signaling pathway plays a key role in regulating cell survival, inflammation, and metabolic modifications in cancer (10). NF-κB p65 upregulates the transcription of PKM2, which in turn activates the NF-κB signaling pathway in cancer cells (11). Emerging evidence suggests crosstalk between metabolic reprogramming and NF-κB-PKM2 positive feedback loop, suggesting their important roles in glioma.

Tripartite motif (TRIM)-containing proteins, a prominent family of E3 ubiquitin ligases, are involved in multiple biological processes, including cell proliferation, programmed cell death, and intracellular signal transduction cascades (12,13). Abnormal expression of TRIM family members has been observed in various cancers, functioning as either oncogenes or tumor suppressors (12-14). TRIM47 was found to be significantly upregulated in hepatocellular carcinoma, intrahepatic cholangiocarcinoma, and osteosarcoma and promotes cancer progression (15-17). TRIM47 regulated metabolic reprogramming to promote thyroid cancer, pancreatic cancer, and hepatocellular carcinoma by ubiquitination of FBP1 (17-19). Importantly, TRIM47 as a tumorigenesis promoter also promoted cell proliferation, cell cycle progression, migration, and invasion of glioma by ubiquitination of p53 and FOXO1 (20,21). However, the specific targets and the mechanisms through which TRIM47 regulates metabolic reprogramming in glioma remain obscure.

In this study, we analyzed TRIM47 expression in glioma tissues and cell lines, evaluated its clinical significance, and explored its effects on glioma cell proliferation, apoptosis, metabolism, and tumorigenesis in vitro and in vivo. We further deciphered the molecular mechanisms underlying TRIM47-mediated glioma progression, focusing on the NF-κB signaling pathway and metabolic reprogramming. We present this article in accordance with the ARRIVE and MDAR reporting checklists (available at https://tcr.amegroups.com/article/view/10.21037/tcr-2026-1-0317/rc).

Methods

Bioinformatics analysis

The TRIM47 gene and protein expression profiles across all tumor samples and normal tissues were evaluated using the The Cancer Genome Atlas (TCGA) and Clinical Proteomic Tumor Analysis Consortium (CPTAC) databases. Gene Expression Profiling Interactive Analysis (GEPIA) was employed to assess the correlation between TRIM47 mRNA expression and survival outcomes in glioma patients. An online analytical tool for investigating gene expression profiles in cancerous and normal tissues is built upon data derived from TCGA database. This study was conducted in accordance with the Declaration of Helsinki and its subsequent amendments.

Immunohistochemistry (IHC)

We acquired glioma tissue microarray (N092Ct01), including glioma tissues and paired normal brain tissues of 46 patients with glioma (age from 20 to 71 years, 30 males and 16 females), from Bioaitech (Xian, China). To ensure a comprehensive assessment of protein expression in the tumors, IHC staining was done strictly following the manufacturer’s instructions (22). Paraffin-embedded sections of tissues were deparaffinized, rehydrated, and underwent antigen retrieval, then incubated with primary antibodies against TRIM47 (26885-1-AP; Proteintech Group, Inc., Rosemont, IL, USA), followed by horseradish peroxidase (HRP)-conjugated anti-rabbit IgG secondary antibodies (8114; Cell Signaling Technology, Danvers, MA, USA, 1:2,000). Two blinded pathologists independently quantified the immunostaining results as previously described (23).

Cell lines and culture

Human glioma cell lines SHG44 (Otwo Biotech, Shenzhen, China, HTX2797), T98G [American Type Culture Collection (ATCC), Manassas, VA, USA, CRL-1690], U251 (Otwo Biotech, HTX1725), U87 (ATCC, HTB-14) and human brain microvascular endothelial cells (HBMEC; ATCC, CRL-3245) were used in this study. Cells were maintained in Dulbecco’s Modified Eagle’s Medium (DMEM) supplemented with 10% fetal bovine serum and 1% penicillin-streptomycin at 37 ℃ in a 5% CO₂ atmosphere.

Knockdown and overexpression of TRIM47

The pLKO.1 lentiviral vector was used to insert shRNA targeting human TRIM47 and scramble shRNA (shNC). For lentiviral production, the pLKO.1-shTRIM47 or pLKO.1-shNC lentiviral vector alongside packaging plasmids (pMD2.G and psPAX2) was co-transfected into 293T cells using Lipofectamine 2000 (Thermo Fisher Scientific, Waltham, MA, USA). Recombinant lentivirus harvested 48 h post-transfection was subsequently used to infect U87 cells. For TRIM47 overexpression, pcDNA3.1 vector containing the full-length cDNA of TRIM47 or blank vector was transfected into T98G cells using Lipofectamine 2000 (Thermo Fisher Scientific). At 48 h after corresponding vector transfection, the cells with high transduction efficiency (>90%) were used for detecting gene knockdown and overexpression efficiency.

RNA extraction and quantitative real-time polymerase chain reaction (qRT-PCR)

RNA was isolated with the TRIzol reagent, and cDNA synthesis was performed using Hifair^® II 1st Strand cDNA Synthesis Kit (Yeasen Biotechnology Co., Ltd, 11119ES60, Shanghai, China). qRT-PCR was conducted utilizing Hieff^® qPCR SYBR Green Master Mix (Yeasen Biotechnology Co., Ltd, 11203ES03). Relative mRNA levels were assessed using the 2⁻^ΔΔCt method, with β-actin as the reference standard. The primer sequences were listed as TRIM47, F: 5'-GTCCAAAGTCCTTGAGCGCC-3', R: 5'-GCTACGGCTGCACTCTTGAT-3'; IKBA, F: 5'-AAGTGATCCGCCAGGTGAAG-3', R: 5'-CTGCTCACAGGCAAGGTGTA-3'; ACTB, F: 5'-CGATTCCTATGTGGGCGACGA-3', R: 5'-TAGGTCTCAAACATGATCTGGGT-3'.

Western blot analysis

Total protein was separated with sodium dodecyl sulfate-polyacrylamide gel electrophoresis and transferred onto a polyvinylidene fluoride membrane. Then, the membranes were incubated with primary antibodies, including TRIM47 (ab72234, 1:1,000), pyruvate kinase M2 (PKM2; ab137791, 1:1,000), NF-κB (ab76311, 1:1,000), p-NF-κB (ab76302, 1:1,000), cleaved caspase-3 (ab32042, 1:1,000), IκBα (ab32518, 1:1,000) and β-actin (ab8227, 1:3,000). After incubation with goat anti-rabbit IgG-HRP (all from Abcam, Cambridge, UK, ab6721, 1:20,000) secondary antibody, bands were visualized using a chemiluminescence system.

Cell viability assay

At 0, 12, 24, and 48 hours post-transfection, the Cell Counting Kit-8 (CCK-8) reagent (Beyotime Biotechnology, Shanghai, China, C0039) was added, and absorbance was measured at 450 nm with a Microplate reader from Bio-Rad (Hercules, CA, USA).

Cell cycle and apoptosis analysis

To analyze the cell cycle and analysis, cells were stained using a Cell Cycle and Apoptosis Analysis Kit (Beyotime Biotechnology, C1052) and analyzed by flow cytometry.

Extracellular flux analysis

Cellular acidification and respiration were measured as extracellular acidification rate (ECAR) and oxygen consumption rate (OCR) using the Seahorse XF Glycolysis Stress Test Kit and the XF Cell Mito Stress Test Kit (Seahorse, 103020-100, 103015-100) as previously described (24). Briefly, cells were seeded at 1×10⁴/well in Seahorse XF cell culture microplates and incubated in an incubator of 37 ℃ and 5% CO₂ for 24 h. Following treatment, culture medium was replaced with Seahorse XF Base Medium, and cells were incubated in a non-CO₂ incubator. Assays were performed using the Seahorse XF Extracellular Flux Analyzer XFe96 (Agilent Technologies, Santa Clara, CA, USA), and the results were analyzed using Wave software (Agilent Technologies).

Glucose uptake and lactate production assays

Using a glucose 2-NBDG assay kit (Biovision, Milpitas, CA, USA, K682-50), glucose uptake was determined. After transfection, cells were subjected to glucose starvation, then treated with Krebs-Ringer Bicarbonate solution and 2-NBDG. Cells were washed and resuspended before undergoing flow cytometry analysis to determine glucose uptake potential with FACScan. Following an overnight incubation, the complete medium was changed to fresh DMEM. After 24 h, the lactate release was determined using Lactic Acid assay kit (Beijing Solarbio Science & Technology Co., BC2230).

Xenograft tumor model

A total of 12 male BALB/c nude mice (aged 6 weeks, 18–22 g) were purchased from Ziyuan Experimental Animal Technology Co., Ltd. (Hangzhou, China) and housed in a specific pathogen-free-grade equipped animal facility with the temperature maintained at 20–25 ℃ and humidity controlled at 50%±5%, following a consistent 12-h dark/12-h light cycle, with free access to autoclaved food and water. After a 7-day acclimatization period in the animal facility, BALB/c nude mice were subcutaneously injected with 5×10⁶ U87 cells transfected with either shNC or shTRIM47-1 (n=6 per group). Tumor size was measured every three days, and the weight was noted after the mice were euthanized. Tumor tissues were formalin-fixed for terminal deoxynucleotidyl transferase dUTP nick-end labeling (TUNEL) assay. The animal experiments were conducted at Affiliated Hospital of Zunyi Medical University. All animal experiments were performed under a project license (No. ZMU21-2301-071) granted by the Ethics Committee of Animal Experiments of the Zunyi Medical University, in compliance with Affiliated Hospital of Zunyi Medical University guidelines for the care and use of animals.

Coimmunoprecipitation and ubiquitination analysis

Cells were lysed with immunoprecipitation lysis buffer. The cell lysates were centrifuged at 4 ℃ for 5 min and then incubated with Protein A/G magnetic beads overnight at 4 ℃ conjugated with anti-TRIM47 (Abcam, ab72234), anti-IκBα (Abcam, ab32518), or control IgG (Santa Cruz Biotechnology, Dallas, TX, USA, sc-515946) antibodies. The beads were then washed, and the immunoprecipitated complexes were analyzed by Western blotting using primary antibodies against TRIM47 (Invitrogen, Carlsbad, CA, USA, MA5-22843), IκBα (Proteintech Group, Inc., 66418-1-Ig) and ubiquitin (Abcam, ab134953).

Protein stability assay

To assess IκBα protein stability, T98G cells with TRIM47 overexpression were treated with the protein synthesis inhibitor cycloheximide (CHX; 0.1 mg/mL; Selleck, Shanghai, China). At 0, 3, and 6 h after treatment, cells were harvested and lysed for Western blot analysis.

Statistical analysis

All the experiments were performed using three (in vitro assays) or six (in vivo assays) independent biological replicates, with technical triplicates for each biological replicate. Data are expressed as mean ± standard deviation (SD) from three independent experiments. No sample size calculation was performed. Blinding and randomization were not performed. Statistical analyses were carried out using GraphPad Prism 8.4.0 (La Jolla, CA, USA). The normality of variable distribution was assessed using the Shapiro-Wilk test, and homogeneity of variances was evaluated using Levene’s test. Student’s t-test or one-way analysis of variance was used to evaluate significance, while the log-rank test was applied for survival analysis. A P value <0.05 was defined as statistically significant.

Results

TRIM47 is upregulated in glioma and correlates with poor prognosis

To clarify TRIM47 expression in glioma, we first used TCGA and CPTAC databases. The results showed that compared to normal tissues, the TRIM47 level was significantly upregulated in GBM tissues (Figure 1A,1B). The analysis using Kaplan-Meier survival curves showed that patients with high TRIM47 expression had a notably shorter overall survival (log-rank P=0.0012) (Figure 1C). The comparison of high versus low TRIM47 expression yielded a hazard ratio of 1.8 (P=0.0014), suggesting that higher TRIM47 level is a negative prognostic indicator for GBM. IHC staining analysis revealed that TRIM47 protein levels were markedly elevated in glioma tissues compared to normal brain tissues (Figure 1D,1E). TRIM47 was similarly overexpressed in glioma cell lines (SHG44, T98G, U251, U87) as opposed to HBMEC (Figure 1F-1H).

Figure 1 TRIM47 expression is increased in glioma tissues and cell lines. (A) The mRNA expression of TRIM47 in 24 kinds of tumors compared to normal tissues from TCGA database. (B) The protein expression of TRIM47 in 10 kinds of tumors compared to normal tissues from CPTAC database. (C) Analysis of the correlation between TRIM47 expression and patient prognosis in glioma using the GEPIA database. (D,E) IHC staining of a glioma tissue microarray analyzing TRIM47 expression in glioma and normal brain tissues (scale bar, 100 µm). (F) qRT-PCR and (G,H) Western blot analysis of TRIM47 expression in multiple glioma cell lines (T98G, U87, U251, SHG44) and human brain microvascular endothelial cells (HBMEC). Data are presented as the means ± SD from three independent experiments. *P<0.05, **P<0.01, ***P<0.001 vs. normal tissue or HBMEC. CPTAC, Clinical Proteomic Tumor Analysis Consortium; GBM, glioblastoma; GEPIA, Gene Expression Profiling Interactive Analysis; HBMEC, human brain microvascular endothelial cells; HR, hazard ratio; IHC, immunohistochemistry; mRNA, messenger RNA; qRT-PCR, quantitative real-time polymerase chain reaction; SD, standard deviation; TCGA, The Cancer Genome Atlas; TPM, transcripts per million.

TRIM47 knockdown suppresses cell growth and metabolic reprogramming and facilitates apoptosis in glioma

To further explore the mechanism by which TRIM47 knockdown reduces glioma cell proliferation, we studied cell viability, cell cycle, apoptosis, and glycolytic metabolism, along with the associated signaling pathways post-TRIM47 interference in U87 cells. qRT-PCR and Western blot confirmed efficient knockdown of TRIM47 by shTRIM47-1, shTRIM47-2, and shTRIM47-3 (Figure 2A-2C). CCK-8 assay results showed that U87 cell viability was notably diminished in the TRIM47 knockdown group after transfection for 24 or 48 h (Figure 2D). Flow cytometry analysis demonstrated that the silencing of TRIM47 induced cell cycle arrest at the G1 phase (Figure 2E,2F) and heightened apoptosis rates (Figure 3A). In U87 cells transfected with shTRIM47-1 or shTRIM47-2, OCR was increased, while ECAR, glucose uptake and lactate production were significantly reduced, compared to the shNC group (Figure 3B-3E). These results imply that the absence of TRIM47 compromises the aerobic glycolytic potential of glioma cells. Western blot demonstrated that TRIM47 knockdown elevated cleaved caspase-3 and lowered PKM2 expression (Figure 3F,3G). Collectively, these data demonstrate that TRIM47 promotes glioma metabolic reprogramming and cell survival by activating the NF-κB pathway and upregulating PKM2 expression.

Figure 2 TRIM47 knockdown inhibits cell viability and cell cycle progression in glioma. (A-C) U87 cells were transfected with lentivirus expressing shRNA targeting TRIM47. The knockdown efficiency was verified by qRT-PCR and Western blot. (D) Cell viability was assessed by the CCK-8 assay. (E,F) Flow cytometry was used to analyze cell cycle distribution. Data are presented as the means ± SD from three independent experiments. **P<0.01, ***P<0.001 vs. shNC. CCK-8, Cell Counting Kit-8; OD, optical density; PI, propidium iodide; qRT-PCR, quantitative real-time polymerase chain reaction; SD, standard deviation; shRNA, short hairpin RNA.

Figure 3 TRIM47 knockdown promotes cell apoptosis and inhibits glycolysis in glioma cells. U87 cells were transfected with lentivirus expressing shRNA targeting TRIM47. (A) Flow cytometry was used to analyze cell apoptosis. (B) OCR and (C) ECAR were measured by Extracellular Flux Analyzer. (D) Cellular glucose uptake capacity was measured by flow cytometry. (E) Lactate production was measured using a biochemical assay. (F,G) Protein levels of NF-κB, p-NF-κB, cleaved-caspase-3, and PKM2 were determined by Western blot. Data are presented as the mean ± SD from three independent experiments. ***P<0.001 vs. shNC. ECAR, extracellular acidification rate; NF-κB, nuclear factor kappa B; OCR, oxygen consumption rate; SD, standard deviation; shRNA, short hairpin RNA.

TRIM47 silencing inhibits tumor growth in vivo

In xenograft tumor models, knockdown of TRIM47 resulted in a significant decrease in both tumor volume and weight compared with the shNC group (Figure 4A-4C). In TRIM47-silenced tumors, TUNEL staining indicated a greater number of apoptotic cells (Figure 4D,4E). Western blot analysis confirmed that p-NF-κB and PKM2 were downregulated, while cleaved caspase-3 was upregulated in the shTRIM47 group (Figure 4F,4G).

Figure 4 TRIM47 knockdown inhibits tumor growth in vivo. A subcutaneous xenograft model was established in nude mice using U87 cells with TRIM47 knockdown. (A) Tumor growth curves were plotted (n=6 mice per group). (B) Representative photographs of excised tumors (n=6 mice per group). (C) Tumor weight was measured (n=6 mice per group). (D,E) Apoptosis in tumor tissues was analyzed by TUNEL staining (scale bar, 50 µm) (n=6 mice per group). (F,G) Protein levels of TRIM47, NF-κB, p-NF-κB, cleaved-caspase-3, and PKM2 in tumor tissues were detected by Western blot (n=3 mice per group). Data are presented as the means ± SD. ***P<0.001 vs. shNC group. NF-κB, nuclear factor kappa B; SD, standard deviation.

TRIM47 overexpression regulates cell growth, apoptosis, and metabolic reprogramming via the NF-κB pathway

To explore the function of TRIM47 and its dependence on NF-κB signaling, we initially verified successful TRIM47 overexpression in T98G cells. Figure 5A-5C illustrates that the levels of TRIM47 mRNA and protein were markedly increased in T98G cells overexpressing TRIM47 when compared to the control groups. The TRIM47 + vehicle group displayed a considerable enhancement in cell viability relative to the vector + vehicle group (Figure 5D). Significantly, this effect was entirely reversed by PDTC, an NF-κB inhibitor, in the TRIM47+PDTC group, indicating that TRIM47-induced cell growth depends on NF-κB activation. TRIM47 overexpression was shown to decrease G1-phase cell arrest and increase the number of cells in the S phase, promoting faster cell cycle progression (Figure 5E,5F). Conversely, PDTC treatment restored G1-phase arrest, abrogating the dynamic changes by TRIM47 overexpression. Apoptosis assays (Figure 6A) further showed that TRIM47 overexpression significantly suppressed apoptotic cell percentages. However, the combination of PDTC and TRIM47 reversed the anti-apoptotic effect, elevating apoptosis rates to the levels found in control groups. TRIM47 overexpression (TRIM47+Vehicle) caused a significant decrease in OCR and rise in ECAR and glucose uptake, effects that were reversed by NF-κB inhibition (Figure 6B-6E). TRIM47 overexpression resulted in elevated lactate production, which PDTC counteracted (Figure 6F). Concurrently, it boosted PKM2 and p-NF-κB expression and lowered cleaved caspase-3 levels, which were inhibited by PDTC (Figure 6G, 6H). In conclusion, TRIM47 boosts glioma metabolic reprogramming and survival via the activation of the NF-κB pathway.

Figure 5 TRIM47 overexpression promotes cell viability and cell cycle progression in glioma via the NF-κB signaling pathway. (A-C) T98G cells were transfected with a TRIM47-overexpressing plasmid. Overexpression efficiency was confirmed by qRT-PCR and Western blot. TRIM47-overexpressing cells were treated with or without 20 µM of the NF-κB-specific inhibitor PDTC. (D) Cell viability was assessed by the CCK-8 assay. (E,F) Flow cytometry was used to analyze cell cycle distribution. Data are presented as the means ± SD from three independent experiments. *P<0.05, **P<0.01, ***P<0.001 vs. vector or vector + vehicle. ^###P<0.001 vs. TRIM47 + vehicle. CCK-8, Cell Counting Kit-8; NF-κB, nuclear factor kappa B; OD, optical density; PI, propidium iodide; qRT-PCR, quantitative real-time polymerase chain reaction; SD, standard deviation.

Figure 6 TRIM47 overexpression inhibits cell apoptosis and promotes glycolysis in glioma via the NF-κB signaling pathway. TRIM47-overexpressing cells were treated with or without 20 µM of the NF-κB-specific inhibitor PDTC. (A) Flow cytometry was used to analyze cell apoptosis. (B) OCR and (C) ECAR were measured by Extracellular Flux Analyzer. (D,E) Cellular glucose uptake capacity was measured by flow cytometry. (F) Lactate production was measured using a biochemical assay. (G,H) Protein levels of NF-κB, p-NF-κB, cleaved-caspase-3, and PKM2 were determined by Western blot. Data are presented as the means ± SD from three independent experiments. ***P<0.001 vs. vector or vector + vehicle. ^###P<0.001 vs. TRIM47 + vehicle. ECAR, extracellular acidification rate; NF-κB, nuclear factor kappa B; OCR, oxygen consumption rate; SD, standard deviation.

TRIM47 interacts with IκBα and induces ubiquitination of IκBα

TRIM47 has been reported to activate the NF-κB signaling by inducing ubiquitination of IκBα in atherosclerosis and gastric cancer (25,26). Therefore, this regulatory mechanism was also determined in glioma cells. As shown in Figure 7A, TRIM47 coimmunoprecipitated with IκBα, and reciprocal immunoprecipitation with anti-IκBα antibody brought down TRIM47 in U87 cells. qRT-PCR revealed no change in IKBA mRNA expression (Figure 7B). However, western blot analysis exhibited that TRIM47 knockdown increased IκBα expression in U87 cells, whereas TRIM47 overexpression suppressed it in T98G cells (Figure 7C,7D), indicating that TRIM47 regulates IκBα at the post-transcriptional level. To further establish whether TRIM47 regulates IκBα protein stability, T98G cells with TRIM47 overexpression were treated with CHX, and the half-life of IκBα was determined. As shown in Figure 7E, IκBα protein stability was significantly decreased in TRIM47-overexpression cells. These findings indicated that TRIM47 destabilizes IκBα in glioma cells. We further performed IκBα immunoprecipitation in TRIM47-overexpression T98G cells to assess IκBα ubiquitination. Immunoblotting with anti-ubiquitin antibody exhibited increased ubiquitination of IκBα in TRIM47-overexpression cells compared to blank vector (Figure 7F). These findings indicate that TRIM47 suppresses IκBα expression by promoting its ubiquitination.

Figure 7 TRIM47 interacts with IκBα and induces ubiquitination of IκBα. (A) Coimmunoprecipitation followed by Western blot confirmed the interaction between TRIM47 and IκBα in U87 cells. IκBα mRNA (B) and protein (C,D) expression levels were examined in U87 cells with TRIM47 knockdown and in T98G cells with TRIM47 overexpression. (E) IκBα protein stability was assessed in T98G cells with TRIM47 overexpression following treatment with 0.1 mg/mL CHX for 0, 3, and 6 h. (F) Ubiquitination of IκBα was evaluated in T98G cells with TRIM47 overexpression. Data are presented as the means ± SD from three independent experiments. **P<0.01, ***P<0.001 vs. shNC or vector. CHX, cycloheximide; IP, immunoprecipitation; mRNA, messenger RNA; SD, standard deviation.

Discussion

The study systematically analyzes the influence of TRIM47 on glioma growth and uncovers its molecular underpinnings, offering novel insights into the cancerous roles of TRIM family proteins in brain tumors. Our significant findings suggest that TRIM47 is abnormally increased in glioma tissues and cell lines, is linked to adverse clinical outcomes in GBM patients, and supports glioma progression through the activation of the NF-κB signaling pathway and metabolic reprogramming.

Glioma, especially GBM, remains a devastating disease with limited therapeutic options and dismal prognosis (27). The identification of reliable prognostic biomarkers and therapeutic targets is critical for improving patient outcomes (28). The study reveals that TRIM47 is expressed at significantly higher levels in glioma tissues compared to normal brain tissues, and increased TRIM47 expression is linked to shorter overall survival in patients with GBM. This research suggests that TRIM47 might serve as a prognostic biomarker for GBM, supporting the stratification of patient risk and the customization of treatment strategies. Also, the role of TRIM47 in glioma progression, as evidenced by both in vitro and in vivo studies, highlights its potential as a target for therapeutic intervention. Inhibiting TRIM47 may provide an alternative path for glioma treatment, particularly for patients with high levels of TRIM47 expression who are expected to have a poor prognosis with standard therapies. Studies found that lowering TRIM47 expression inhibits glioma cell viability, causes G1 phase cell cycle arrest, and promotes apoptosis, while higher TRIM47 expression produces the opposite results. The results are consistent with earlier findings that TRIM47 behaves as an oncogene in other cancers, such as hepatocellular carcinoma, intrahepatic cholangiocarcinoma and osteosarcoma, by promoting cell proliferation and inhibiting apoptosis (15-17). Loss of TRIM47 results in activation of p21 and inhibition of cyclin D1 expression, attenuating cell-cycle progression in GBM (20,21). TRIM47 also regulated expression of CDK6, CDK4, cyclin D1, cleaved PARP, cleaved caspase-3, and cleaved caspase-9 to promote cell cycle and restrain apoptosis in head and neck squamous cell carcinoma (29). Our results showed that TRIM47 knockdown promoted cleaved caspase-3 expression while TRIM47 overexpression inhibited it in glioma cells.

Metabolic reprogramming, characterized by the preference for aerobic glycolysis even in oxygen-rich environments, is a well-established hallmark of cancer cells (30-32). The metabolic alteration supplies cancer cells with ample energy and biosynthetic materials necessary for their fast proliferation, invasion, and endurance in the tumor microenvironment (33-35). PKM2 is a significant enzyme in glycolysis that aids the Warburg effect by catalyzing the last phase of glycolysis, resulting in the creation of pyruvate and adenosine triphosphate (7-9). Our study shows that TRIM47 modulates PKM2 expression, and TRIM47 knockdown reduces glucose uptake and lactate production in glioma cells, indicating that TRIM47 regulates glycolytic metabolism through PKM2. This finding is consistent with previous reports that PKM2 is overexpressed in glioma and contributes to tumor progression (36,37). The regulation of PKM2 by TRIM47 may be a critical mechanism underlying metabolic reprogramming in glioma, providing a link between TRIM47 and the metabolic adaptation of cancer cells.

The NF-κB pathway is indispensable for the modulation of multiple biological functions, such as cell survival, growth, inflammatory responses and metabolic processes (10,38). Aberrant activation of the NF-κB pathway is a common occurrence in gliomas, and this dysregulated signaling cascade plays a pivotal role in driving tumor development and mediating therapy resistance (39,40). The findings suggest that TRIM47 triggers the NF-κB signaling pathway, as indicated by increased NF-κB phosphorylation when TRIM47 is overexpressed and decreased phosphorylation when TRIM47 is knocked down. The tumorigenic impacts of TRIM47 overexpression on cell viability, cell cycle progression, apoptosis, and metabolic reprogramming can also be mitigated by the NF-κB inhibitor PDTC. The findings indicate that the NF-κB signaling pathway functions as a downstream mediator of TRIM47 in glioma. Emerging evidence suggests that metabolic reprogramming and NF-κB activation are closely interconnected in cancer cells (41,42). The NF-κB pathway can influence metabolic enzymes and transporters to enhance the Warburg effect, and metabolic intermediates can affect NF-κB activation through different mechanisms (43,44). Our research indicates that the upregulation of PKM2 expression induced by TRIM47 overexpression was inhibited by NF-κB inhibitor PDTC, suggesting that PKM2 is regulated by TRIM47 via the NF-κB signaling pathway.

IκBα is a major inhibitor of NF-κB protein, which is degraded when the organism is stimulated, and the free NF-κB complex translocates to the nucleus and mediates transcription of various target genes. TRIM47 has been reported to activate the NF-κB signaling by inducing ubiquitination of IκBα in atherosclerosis and gastric cancer (25,26). In line with these findings, we found that TRIM47 decreased IκBα protein stability and increased IκBα ubiquitination level, suggesting that TRIM47 may regulate NF-κB signaling pathway via IκBα ubiquitination in glioma.

There are several limitations in this study. First, the involvement of IκBα-NF-κB regulatory axis in the functions of TRIM47 in glioma requires further investigation through in-depth experimental research. Second, although IκBα is a major upstream factor of NF-κB protein, PKM2 is also involved in the regulation of the NF-κB signaling pathway in cancer cells, which in turn upregulates transcription of PKM2 (11). Therefore, whether a positive feedback loop of PKM2-NF-κB signaling pathway is involved in the carcinogenesis of TRIM47 in glioma should be further addressed. Additionally, the reliance on a relatively small commercial tissue microarray limits the generalizability of the IHC findings. Therefore, future research should validate the expression of TRIM47 with a larger, adequately powered sample size. More precise TRIM47-targeting strategies, such as proteolysis-targeting chimeras and nanoparticle-based delivery systems, need further investigation in future studies. Finally, the potential of TRIM47 as a therapeutic target for glioma necessitates further drug screening and clinical studies.

Conclusions

In conclusion, our study established that increased TRIM47 in glioma is linked to poor prognosis and promotes tumor progression by activating the NF-κB signaling pathway and affecting glycolytic metabolism via regulating PKM2 expression. Targeting TRIM47 might serve as a potential treatment for glioma. The study reveals new insights into the molecular mechanisms behind glioma progression and suggests a promising target for the development of new therapeutic strategies for treating glioma.

Acknowledgments

None.

Footnote

Reporting Checklist: The authors have completed the ARRIVE and MDAR reporting checklists. Available at https://tcr.amegroups.com/article/view/10.21037/tcr-2026-1-0317/rc

Data Sharing Statement: Available at https://tcr.amegroups.com/article/view/10.21037/tcr-2026-1-0317/dss

Peer Review File: Available at https://tcr.amegroups.com/article/view/10.21037/tcr-2026-1-0317/prf

Funding: None.

Conflicts of Interest: All authors have completed the ICMJE uniform disclosure form (available at https://tcr.amegroups.com/article/view/10.21037/tcr-2026-1-0317/coif). The authors have no conflicts of interest to declare.

Ethical Statement: The authors are accountable for all aspects of the work in ensuring that questions related to the accuracy or integrity of any part of the work are appropriately investigated and resolved. This study was conducted in accordance with the Declaration of Helsinki and its subsequent amendments. The animal experiments were conducted at Affiliated Hospital of Zunyi Medical University. All animal experiments were performed under a project license (No. ZMU21-2301-071) granted by the Ethics Committee of Animal Experiments of the Zunyi Medical University, in compliance with Affiliated Hospital of Zunyi Medical University guidelines for the care and use of animals.

Open Access Statement: This is an Open Access article distributed in accordance with the Creative Commons Attribution-NonCommercial-NoDerivs 4.0 International License (CC BY-NC-ND 4.0), which permits the non-commercial replication and distribution of the article with the strict proviso that no changes or edits are made and the original work is properly cited (including links to both the formal publication through the relevant DOI and the license). See: https://creativecommons.org/licenses/by-nc-nd/4.0/.

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