Journal: Cell Death & Disease

Article Title: The IGF2BP3–FASN axis drives lipid metabolic reprogramming to promote brain colonization in non-small cell lung cancer

doi: 10.1038/s41419-025-08006-z

Figure Lengend Snippet: A IGF2BP3 expression was elevated in N2 ( n = 74) and N3 ( n = 2) stage metastatic tumor compared with N1 stage metastatic tumor ( n = 95) in lung adenocarcinoma (LUAD) patients with lymph node metastasis according to TCGA database. Data are presented as median value. B IGF2BP3 expression levels were elevated in brain metastatic tumors (BT) ( n = 6) compared with primary lung tumors (LT) ( n = 8) in early passage cell lines derived from primary patient samples of lung-to-brain metastases according to GSE110495 dataset. Data are presented as median value. C IGF2BP3 expression levels were elevated in LLC-derived brain metastatic cells (Brm) compared with primary cells according to GSE83132 dataset. Data are presented as mean ± SD; n = 3. D Overall survival analysis of LUAD patients at the N1 stage based on TCGA project was calculated with the log-rank test and stratified by high or low IGF2BP3 expression (high: top 50%, n = 127; low: bottom 50%, n = 126). E – H Pearson correlation analysis between IGF2BP3 expression and key EMT markers (TGFB1, SNAI2, N-cadherin, and MMP9) in LUAD patients according to GSE33532 dataset ( n = 100). I Schematic of the subcutaneous tumor and stereotactic brain injection tumor model. J Hematoxylin and eosin (H&E) staining and immunohistochemistry (IHC) analysis of IGF2BP3 and MMP9 in representative subcutaneous tumor and xenograft tumor in the brain from mice. Scale bar = 20 μm. K Quantification of IGF2BP3 and MMP9 expression levels from IHC analysis in J using Image Pro Plus. Data are presented as mean ± SD; n = 3. * P < 0.05; ** P < 0.01; **** P < 0.0001 (two-tailed t -test).

Article Snippet: Anti-IGF2BP3 mouse monoclonal antibody (EM1701-15, HuaBio, China) and anti-IgG mouse monoclonal antibody were conjugated to magnetic beads to immunoprecipitate RNA-protein complexes.

Techniques: Expressing, Derivative Assay, Injection, Staining, Immunohistochemistry, Two Tailed Test

Journal: Cell Death & Disease

Article Title: The IGF2BP3–FASN axis drives lipid metabolic reprogramming to promote brain colonization in non-small cell lung cancer

doi: 10.1038/s41419-025-08006-z

Figure Lengend Snippet: A Wound healing assay showing cell migration ability in H1299 cells treated with shRNAs targeting IGF2BP3 or control (shCtrl) after 24 h. Scale bars = 50 μm. B Quantification of cell migration in IGF2BP3-knockdown H1299 cells from ( A ) using ImageJ. Data are presented as mean ± SD; n = 3. C Transwell invasion assay showing cell invasion ability in H1299 cells treated with shRNAs targeting IGF2BP3 or control after 24 h. Scale bars = 10 μm. D Quantification of cell invasion in IGF2BP3-knockdown H1299 cells from ( C ) using ImageJ. Data are presented as mean ± SD; n = 3. E Wound healing assay showing cell migration ability in H1299 cells overexpressing IGF2BP3 (OE-IGF2BP3) or control after 24 h. Scale bars = 50 μm. F Quantification of cell migration in IGF2BP3-overexpressing H1299 cells from ( E ) using ImageJ. Data are presented as mean ± SD; n = 3. G Transwell invasion assay showing cell invasion ability in H1299 cells overexpressing IGF2BP3 or control after 24 h. Scale bars = 10= μm. H Quantification of cell invasion in IGF2BP3-overexpressing H1299 cells from ( G ) using ImageJ. Data are presented as mean ± SD; n = 3. I – K . H1299-luc cells (5 × 10 5 ) with IGF2BP3 knockout (sgIGF2BP3) or control (sgCtrl) were injected into BALB/c-nu/nu mice via stereotactic brain injection ( n = 6 per group). Representative bioluminescent images of NSCLC xenograft tumor in the brain captured by the Neurostar IVIS preclinical in vivo imaging system on day 27 are shown in ( I , J ). Quantification of bioluminescence in the brain on day 27 is shown in ( K ). Data are presented as mean ± SD; n = 6. L – N H1299-luc cells (5 × 10 5 ) overexpressing IGF2BP3 (OE-IGF2BP3) or control were injected into BALB/c-nu/nu mice via stereotactic brain injection ( n = 6 per group). Representative bioluminescent images of NSCLC xenograft tumor in the brain on day 27 are shown in ( L , M ). Quantification of bioluminescence in the brain on day 27 is shown in ( N ). Data are presented as mean ± SD; n = 6. O H&E staining and IHC analysis of IGF2BP3 and MMP9 in representative brain tumors from IGF2BP3-knockout and control mice. Scale bar = 20 μm. P Quantification of IGF2BP3 and MMP9 expression levels from IHC analysis in ( O ) using Image Pro Plus. Data are presented as mean ± SD; n = 3. Q IHC staining depicted the correlation between IGF2BP3 and MMP9 expression in brain tumors from IGF2BP3-knockout and control mice ( n = 3). R H&E staining and IHC analysis of IGF2BP3 and MMP9 in representative brain tumors from IGF2BP3-overexpressing and control mice. Scale bar = 20 μm. S Quantification of IGF2BP3 and MMP9 expression levels from IHC analysis in ( R ) using Image Pro Plus. Data are presented as mean ± SD; n = 3. T IHC staining depicted the correlation between IGF2BP3 and MMP9 expression in brain tumors from IGF2BP3-overexpressing and control mice ( n = 3). * P < 0.05; ** P < 0.01; *** P < 0.001; **** P < 0.0001 (two-tailed t -test).

Article Snippet: Anti-IGF2BP3 mouse monoclonal antibody (EM1701-15, HuaBio, China) and anti-IgG mouse monoclonal antibody were conjugated to magnetic beads to immunoprecipitate RNA-protein complexes.

Techniques: Wound Healing Assay, Migration, Control, Knockdown, Transwell Invasion Assay, Knock-Out, Injection, In Vivo Imaging, Staining, Expressing, Immunohistochemistry, Two Tailed Test

Journal: Cell Death & Disease

Article Title: The IGF2BP3–FASN axis drives lipid metabolic reprogramming to promote brain colonization in non-small cell lung cancer

doi: 10.1038/s41419-025-08006-z

Figure Lengend Snippet: A Volcano plot of differentially expressed genes (fold change > 1.5, p < 0.05) in H1299 cells stably expressing shRNAs targeting IGF2BP3 (shIGF2BP3) or scramble control (shCtrl). B Heatmap of differentially expressed genes (fold change > 1.5, p < 0.05) between IGF2BP3-knockdown and control H1299 cells measured by z -score. The color means the z score of cluster analysis ( C ) KEGG enrichment analysis of differentially expressed genes (fold change > 1.5, p < 0.05) identified by RNA-seq in IGF2BP3-knockdown H1299 cells. The color means −log10 ( p value) and the dot size means gene count of different pathways. D Gene Set Enrichment Analysis (GSEA) showing decreased fatty acid metabolism in IGF2BP3-knockdown cells. E , F Principal component analysis (PCA) and orthogonal partial least squares-discriminant analysis (OPLS-DA) of lipidomics data in IGF2BP3-knockdown H1299 cells. G Volcano plot of differential metabolites (fold change > 1.5, p < 0.05) in IGF2BP3-knockdown H1299 cells. H KEGG enrichment analysis of differential metabolites (fold change > 1.5, p < 0.05) identified by lipidomics in IGF2BP3-knockdown H1299 cells. The color means p value and dot size means gene count of pathways. I Heatmap of differential metabolites (fold change > 1.5, p < 0.05) between IGF2BP3-knockdown and control H1299 cells measured by z -score. The color means the z score of cluster analysis. J , K Neutral lipid levels detected by BODIPY 493/503 staining in IGF2BP3-knockout brain tumors examined by microscope. Representative fluorescent images ( J ) and quantification ( K ) are shown. Data are presented as mean ± SD; n = 3. *** P < 0.001 (two-tailed t -test).

Article Snippet: Anti-IGF2BP3 mouse monoclonal antibody (EM1701-15, HuaBio, China) and anti-IgG mouse monoclonal antibody were conjugated to magnetic beads to immunoprecipitate RNA-protein complexes.

Techniques: Stable Transfection, Expressing, Control, Knockdown, RNA Sequencing, Staining, Knock-Out, Microscopy, Two Tailed Test

Journal: Cell Death & Disease

Article Title: The IGF2BP3–FASN axis drives lipid metabolic reprogramming to promote brain colonization in non-small cell lung cancer

doi: 10.1038/s41419-025-08006-z

Figure Lengend Snippet: A Total triglycerides assay kit showing total triglyceride levels were decreased in IGF2BP3-knockdown H1299 cells compared with control cells. Data are presented as mean ± SD; n = 3. B Total cholesterol assay kit showing total cholesterol levels were decreased in IGF2BP3-knockdown H1299 cells compared with control cells. Data are presented as mean ± SD; n = 3. C Total triglycerides assay kit showing total triglyceride levels were elevated in IGF2BP3-overexpressing H1299 cells compared with control cells. Data are presented as mean ± SD; n = 3. D Total cholesterol assay kit showing total cholesterol levels were elevated in IGF2BP3-overexpressing H1299 cells compared with control cells. Data are presented as mean ± SD; n = 3. E , F Neutral lipid levels detected by BODIPY 493/503 staining in IGF2BP3-knockdown H1299 cells. Representative fluorescent images ( E ) and quantification ( F ) are shown. Data are presented as mean ± SD; n = 3. G , H Neutral lipid levels measured by Nile Red staining in IGF2BP3-knockdown H1299 cells. Representative fluorescent images ( G ) and quantification ( H ) are shown. Data are presented as mean ± SD; n = 3. I , J Neutral lipid levels detected by BODIPY 493/503 staining in IGF2BP3-overexpressing H1299 cells. Representative fluorescent images ( I ) and quantification ( J ) are shown. Data are presented as mean ± SD; n = 3. K , L Neutral lipid levels measured by Nile Red staining in IGF2BP3-overexpressing H1299 cells. Representative fluorescent images ( K ) and quantification ( L ) are shown. Data are presented as mean ± SD; n = 3. M , N Neutral lipid levels analyzed by Nile Red staining using flow cytometry in IGF2BP3-knockdown H1299 cells. Representative images ( M ) and quantification ( N ) are shown. Data are presented as mean ± SD; n = 3. O , P Neutral lipid levels analyzed by Nile Red staining using flow cytometry in IGF2BP3-overexpressing H1299 cells. Representative images ( O ) and quantification ( P ) are shown. Data are presented as mean ± SD; n = 3. * P < 0.05; ** P < 0.01; *** P < 0.001; **** P < 0.0001 (two-tailed t -test).

Article Snippet: Anti-IGF2BP3 mouse monoclonal antibody (EM1701-15, HuaBio, China) and anti-IgG mouse monoclonal antibody were conjugated to magnetic beads to immunoprecipitate RNA-protein complexes.

Techniques: Knockdown, Control, Cholesterol Assay, Staining, Flow Cytometry, Two Tailed Test

Journal: Cell Death & Disease

Article Title: The IGF2BP3–FASN axis drives lipid metabolic reprogramming to promote brain colonization in non-small cell lung cancer

doi: 10.1038/s41419-025-08006-z

Figure Lengend Snippet: A RT-qPCR analysis of fatty acid metabolism-related genes with cDNA microarrays in IGF2BP3-knockdown H1299 cells ( n = 3). The color means the relative mRNA expression level of indicated genes. B Overlapping analysis of IGF2BP3-regulated genes detected by RIP-seq in GSE90639 , GSE79147 , GSE230646 , and GSE231938 (fold change > 1.5, p < 0.05). C Overlapping analysis of genes from ( A ) and ( B ). D RIP-seq data showing potential binding of FASN mRNA to IGF2BP3 according to GSE231938 dataset. E RIP assay in IGF2BP3-overexpressing H1299 cells were verified by Western blotting. F RIP-qPCR analysis of FASN mRNA enrichment in IGF2BP3-overexpressing H1299 cells. Data are presented as mean ± SD; n = 3. G Pearson correlation analysis of IGF2BP3 and FASN expression in metastasis tumor tissue of LUAD patients according to GSE198291 dataset ( n = 312). H , I FASN mRNA expression evaluated by RT-qPCR in IGF2BP3-knockdown H1299 and H157 cells. Data are presented as mean ± SD; n = 3. J , K FASN protein expression evaluated by Western blotting in IGF2BP3-knockdown H1299 and H157 cells. L FASN mRNA half-life assessed by RT-qPCR in IGF2BP3-knockdown H1299 cells treated with 20 μg/mL actinomycin D for 0, 1, 3, 6, 9, 16, and 24 hours. Data are presented as mean ± SD; n = 3. M FASN mRNA expression evaluated by RT-qPCR in shCtrl and shMETTL3 H1299 cells. Data are presented as mean ± SD; n = 3. N FASN and IGF2BP3 protein levels evaluated by Western blotting in shCtrl and shMETTL3 H1299 cells. O FASN mRNA half-life assessed by RT-qPCR in METTL3-knockdown H1299 cells treated with 20 μg/mL actinomycin D for 0, 1, 3, 6, 9, 16, and 24 h. Data are presented as mean ± SD; n = 3. * P < 0.05; ** P < 0.01; *** P < 0.001; **** P < 0.0001 (two-tailed t -test).

Article Snippet: Anti-IGF2BP3 mouse monoclonal antibody (EM1701-15, HuaBio, China) and anti-IgG mouse monoclonal antibody were conjugated to magnetic beads to immunoprecipitate RNA-protein complexes.

Techniques: Quantitative RT-PCR, Knockdown, Expressing, Binding Assay, Western Blot, Two Tailed Test

Journal: Cell Death & Disease

Article Title: The IGF2BP3–FASN axis drives lipid metabolic reprogramming to promote brain colonization in non-small cell lung cancer

doi: 10.1038/s41419-025-08006-z

Figure Lengend Snippet: A , B IGF2BP3 and FASN mRNA expression evaluated by RT-qPCR in IGF2BP3-knockdown and FASN-overexpressing H1299 and H157 cells. Data are presented as mean ± SD; n = 3. C , D IGF2BP3 and FASN protein levels evaluated by Western blotting in IGF2BP3-knockdown and FASN-overexpressing H1299 and H157 cells. E Wound healing assay showing cell migration ability in H1299 cells treated with IGF2BP3 knockdown and FASN overexpression for 24 h. Scale bars = 50 μm. F Quantification of cell migration in IGF2BP3-knockdown and FASN-overexpressing H1299 cells from ( E ) using ImageJ. Data are presented as mean ± SD; n = 3. G Wound healing assay showing cell migration ability in H157 cells treated with IGF2BP3 knockdown and FASN overexpression for 24 h. Scale bars = 50 μm. H Quantification of cell migration in IGF2BP3-knockdown and FASN-overexpressing H157 cells from ( G ) using ImageJ. Data are presented as mean ± SD; n = 3. I Transwell invasion assay showing invasion ability in H1299 cells treated with IGF2BP3 knockdown and FASN overexpression for 24 h. Scale bars = 10 μm. J Quantification of cell invasion in IGF2BP3-knockdown H1299 cells from ( I ) using ImageJ. Data are presented as mean ± SD; n = 3. K Transwell invasion assay showing invasion ability in H157 cells treated with IGF2BP3 knockdown and FASN overexpression for 24 h. Scale bars = 10 μm. L Quantification of cell invasion in IGF2BP3-knockdown H157 cells from ( K ) using ImageJ. Data are presented as mean ± SD; n = 3. M Total triglycerides assay kit showing total triglyceride levels were rescued in IGF2BP3-knockdown and FASN-overexpressing H1299 cells compared with IGF2BP3-knockdown cells. Data are presented as mean ± SD; n = 3. N Total cholesterol assay kit showing total cholesterol levels were rescued in IGF2BP3-knockdown and FASN-overexpressing H1299 cells compared with IGF2BP3-knockdown cells. Data are presented as mean ± SD; n = 3. O Total triglycerides assay kit showing total triglyceride levels were rescued in IGF2BP3-knockdown and FASN-overexpressing H157 cells compared with IGF2BP3-knockdown cells. Data are presented as mean ± SD; n = 3. P Total cholesterol assay kit showing total cholesterol levels were rescued in IGF2BP3-knockdown and FASN-overexpressing H157 cells compared with IGF2BP3-knockdown cells. Data are presented as mean ± SD; n = 3. Q – T Neutral lipid levels analyzed by flow cytometry in IGF2BP3-knockdown and FASN-overexpressing H1299 and H157 cells stained with Nile Red. Representative images ( Q , S ) and quantification ( R , T ) are shown. Data are presented as mean ± SD; n = 3. U , V Neutral lipid levels detected by BODIPY 493/503 staining in IGF2BP3-knockdown and FASN-overexpressing H1299 cells. Representative fluorescent images ( U ) and quantification ( V ) are shown. Data are presented as mean ± SD; n = 3. W , X Neutral lipid levels measured by Nile Red staining in IGF2BP3-knockdown and FASN-overexpressing H1299 cells. Representative fluorescent images ( W ) and quantification ( X ) are shown. Data are presented as mean ± SD; n = 3. * P < 0.05; ** P < 0.01; *** P < 0.001; **** P < 0.0001 (two-tailed t -test).

Article Snippet: Anti-IGF2BP3 mouse monoclonal antibody (EM1701-15, HuaBio, China) and anti-IgG mouse monoclonal antibody were conjugated to magnetic beads to immunoprecipitate RNA-protein complexes.

Techniques: Expressing, Quantitative RT-PCR, Knockdown, Western Blot, Wound Healing Assay, Migration, Over Expression, Transwell Invasion Assay, Cholesterol Assay, Flow Cytometry, Staining, Two Tailed Test

Journal: Cell Death & Disease

Article Title: The IGF2BP3–FASN axis drives lipid metabolic reprogramming to promote brain colonization in non-small cell lung cancer

doi: 10.1038/s41419-025-08006-z

Figure Lengend Snippet: A FASN expression was elevated in LLC1-derived leptomeningeal metastatic cells (Lep) compared with primary cells according to GSE83132 dataset. Data are presented as mean ± SD; n = 3. B FASN expression was elevated in PC9-derived brain metastatic cells (Brm) compared with primary cells according to GSE83132 dataset. Data are presented as mean ± SD; n = 3. C Overall survival of metastatic lung cancer patients was calculated with the log-rank test and stratified by high or low FASN expression (high: top 50%, n = 27; low: bottom 50%, n = 26; GSE30219 ). D IGF2BP3 and FASN protein expression detected by Western blotting in IGF2BP3-knockout and FASN-overexpressing H1299-luc cells. E , F H1299-luc cells (5 × 10 5 ) with IGF2BP3 knockout and FASN overexpression were injected into BALB/c-nu/nu mice via stereotactic brain injection ( n = 6 per group). Representative bioluminescent images of NSCLC xenograft tumor in the brain on day 27 are shown in ( E ). Quantification of bioluminescence on day 27 is shown in ( F ). Data are presented as mean ± SD; n = 6. G , H Representative bioluminescent images of brain tumors on day 27 were captured by the Neurostar IVIS preclinical in vivo imaging system ( G ). Quantification of bioluminescence in brain tumors on day 27 is shown in ( H ). Data are presented as mean ± SD; n = 6. I H&E staining and IHC analysis of IGF2BP3 and MMP9 in representative brain tumors from IGF2BP3-knockout and FASN-overexpressing mice. Scale bar = 20 μm. J Quantification of IGF2BP3 and MMP9 expression levels from IHC analysis in ( I ) using Image Pro Plus. Data are presented as mean ± SD; n = 3. K , L Neutral lipid levels detected by BODIPY 493/503 staining in representative brain tumors from IGF2BP3-knockout and FASN-overexpressing mice. Representative fluorescent images ( K ) and quantification ( L ) are shown. Data are presented as mean ± SD; n = 3. * P < 0.05; ** P < 0.01; **** P < 0.0001 (two-tailed t -test).

Article Snippet: Anti-IGF2BP3 mouse monoclonal antibody (EM1701-15, HuaBio, China) and anti-IgG mouse monoclonal antibody were conjugated to magnetic beads to immunoprecipitate RNA-protein complexes.

Techniques: Expressing, Derivative Assay, Western Blot, Knock-Out, Over Expression, Injection, In Vivo Imaging, Staining, Two Tailed Test