Journal: Journal of Experimental & Clinical Cancer Research : CR

Article Title: Novel MAFG-METTL14-SCD1 axis regulates lipid metabolism mediating choroidal melanoma distant metastasis

doi: 10.1186/s13046-025-03595-1

Figure Lengend Snippet: METTL14 mediates lipid metabolism component alterations through SCD1 to promote the characteristics of choroidal melanoma metastasis. a , Venn diagram and pathway enrichment analysis identifying SCD1 , HSD17B8 , ELOVL3 , ACSL6 , ACSBG1 , ACADL , and ACAA2 as downstream target candidates of METTL14. b , RM2target ( http://rm2target.canceromics.org/ ) showing the correlation between SCD1 and METTL14 in various cancers. c , Volcano plot showing 315 downregulated and 259 upregulated differential metabolites in SCD1-treated CM cells (MUM-2B) (FC > 1.5, P < 0.05) compared to the control group. d , KEGG enrichment analysis of metabolism pathways associated with differential metabolites. e , Peak area values of Behenic acid and Stearic acid in the unsaturated fatty acid biosynthesis pathway, comparing the shNC group vs. shSCD1 group. f , Free fatty acid colorimetry (NEFA/FFA) showing an increase in free fatty acid content after silencing SCD1. g , Flow cytometry showing ROS levels after silencing SCD1. h-i , Representative coronal and axial sectional PET images showing the uptake of 18 F-FDG in the lung and liver of mice in two different groups (shNC and shSCD1, n = 5 for each). j-k , Representative images of excised lungs and livers from mice injected with the indicated cells. (Scale bar, 1 cm). l , Representative hematoxylin and eosin (H&E) staining images of lung and liver tissues. m , Immunohistochemical analysis of mouse lung and liver tissues with an anti-SCD1 antibody in two different groups (shNC and shSCD1, n = 5 for each). Scale bar, 50 μm. n , Representative ORO staining images of frozen mouse lung and liver tissues

Article Snippet: Subsequently, the samples were maintained at 4 °C overnight with primary antibodies against METTL14 (dilution 1:1000, HPA038002, Sigma-Aldrich), SCD1 (dilution 1:500, 2794 S, CST), and MAFG (dilution 1:500, #ab154318, Abcam).

Techniques: Control, Colorimetric Assay, Flow Cytometry, Injection, Staining, Immunohistochemical staining

Journal: Journal of Experimental & Clinical Cancer Research : CR

Article Title: Novel MAFG-METTL14-SCD1 axis regulates lipid metabolism mediating choroidal melanoma distant metastasis

doi: 10.1186/s13046-025-03595-1

Figure Lengend Snippet: METTL14 mediates choroidal melanoma lipid metabolism by inducing SCD1 m 6 A methylation modification. a , Schematic diagram illustrating the METTL14-SCD1 regulatory axis via a specific m 6 A modification-dependent mechanism. b , The SRAMP ( http://www.cuilab.cn/sramp ) predicting the distribution of high-confidence m 6 A methylation sites and scores evaluation for SCD1 mRNA. c , Linear correlation between METTL14 expression and SCD1 expression. d , qRT-PCR analysis showing alterations in SCD1 mRNA levels after METTL14 depletion. e , FISH assay showing the expression and co-localization of METTL14 (green) with SCD1 (red) in CM cells. DAPI (blue) is used as the nuclear marker. Scale bar, 5 μm. f , m 6 A-MeRIP-qPCR assay showing enrichment of SCD1 by m 6 A-specific antibodies. g , ENCORI-CLIP database ( http://starbase.sysu.edu.cn/ ) illustrating the RBP-mRNA interactions between METTL14 and SCD1. h , Double luciferase assay confirming METTL14 directly binds to the 2492 site of SCD1 3’UTR, with mutations at this site. i , Representative western blot images showing SCD1 expression in normal choroidal tissues and CM tissues. j , Immunohistochemical analysis of human normal choroidal and CM tissue samples with the indicated antibodies. Representative images shown. Scale bar, 100 μm. (Normal: n = 10, CM: n = 34). k , Membrane fluidity assay showing changes in cell membrane fluidity after interfering with SCD1 expression in C918 and MUM-2B cells. l , Transmission electron microscopy (TEM) showing lipid droplet reduction and mitochondrial contraction following the silencing of SCD1 in MUM-2B cells. LD, lipid droplet; M, mitochondria. Scale bars, 1 μm

Article Snippet: Subsequently, the samples were maintained at 4 °C overnight with primary antibodies against METTL14 (dilution 1:1000, HPA038002, Sigma-Aldrich), SCD1 (dilution 1:500, 2794 S, CST), and MAFG (dilution 1:500, #ab154318, Abcam).

Techniques: Methylation, Modification, Expressing, Quantitative RT-PCR, Marker, Luciferase, Western Blot, Immunohistochemical staining, Membrane, Transmission Assay, Electron Microscopy

Journal: Journal of Experimental & Clinical Cancer Research : CR

Article Title: Novel MAFG-METTL14-SCD1 axis regulates lipid metabolism mediating choroidal melanoma distant metastasis

doi: 10.1186/s13046-025-03595-1

Figure Lengend Snippet: The METTL14-SCD1 signaling axis regulates lipid metabolism to promote CM invasion and metastasis. a , Radar chart illustrating the top 10 downregulated differential metabolic products obtained from the shNC + shMETTL14 group vs. shNC + shSCD1 group ( P < 0.05). b , Rescue experiments determining cell membrane fluidity by overexpressing SCD1 upon silencing METTL14 in C918 and MUM-2B cells. c , Rescue experiments revealing free fatty acid content treated by overexpression of SCD1 upon silencing METTL14 in MUM-2B cells. d , Flow cytometry detecting ROS levels in the recovery experiment groups. e , Rescue experiments determining the number of lipid droplets after overexpression of SCD1 upon silencing METTL14 in MUM-2B cells. LD, lipid droplet. Scale bars, 5 μm and 1 μm. f-g , Representative coronal and axial sectional PET images showing the uptake of 18 F-FDG in the lung and liver of mice in three different groups (shNC + pcDH, shM14 + pcDH, and shM14 + SCD1). h , Representative images of excised lungs and livers from mice injected with the indicated cells. Scale bar, 1 cm. i , Representative images of H&E staining of lung and liver tissues. j , Immunohistochemical analysis of mouse lung and liver tissues performed with an anti-METTL14 antibody in three different groups (shNC + pcDH, shM14 + pcDH, and shM14 + SCD1). Scale bars, 50 μm. k , Representative ORO staining images of lung and liver tissues

Article Snippet: Subsequently, the samples were maintained at 4 °C overnight with primary antibodies against METTL14 (dilution 1:1000, HPA038002, Sigma-Aldrich), SCD1 (dilution 1:500, 2794 S, CST), and MAFG (dilution 1:500, #ab154318, Abcam).

Techniques: Membrane, Over Expression, Flow Cytometry, Injection, Staining, Immunohistochemical staining

Journal: Journal of Experimental & Clinical Cancer Research : CR

Article Title: Novel MAFG-METTL14-SCD1 axis regulates lipid metabolism mediating choroidal melanoma distant metastasis

doi: 10.1186/s13046-025-03595-1

Figure Lengend Snippet: SCD1 inhibitor aramchol exhibits diet-dependent anti-metastatic effects. a , Representative multiplexed immunohistochemistry staining images of human normal choroidal tissue and CM tissue samples showing METTL14 (yellow), MAFG (purple), SCD1 (green), and DAPI (blue). b , Schematic representation of the therapy schedule. Mice were acclimated to the cages for 3 days, during which they were all fed normal chow (brown-color pellets). Following acclimation, the mice were divided into three dietary groups: normal-fat diet (NFD), high-fat diet rich in stearate (S-HFD), and high-fat diet rich in oleic acid (O-HFD). The interventions began 3 days prior to injection and continued throughout the study ( n = 5). c , Representative coronal and axial sectional PET images showing the uptake of 18 F-FDG in the lung of mice in four different groups. d , Representative images of excised lungs from mice injected with the indicated cells. Scale bar, 1 cm. e , Representative H&E staining images of lung tissues

Article Snippet: Subsequently, the samples were maintained at 4 °C overnight with primary antibodies against METTL14 (dilution 1:1000, HPA038002, Sigma-Aldrich), SCD1 (dilution 1:500, 2794 S, CST), and MAFG (dilution 1:500, #ab154318, Abcam).

Techniques: Immunohistochemistry, Staining, Injection