Journal: BMC Cancer

Article Title: RBMS1-HSPA8 axis activation drives head and neck squamous cell carcinoma progression

doi: 10.1186/s12885-025-13937-z

Figure Lengend Snippet: RBMS1 expression in four HNSCC datasets and validation in HNSCC tissue. A Expression of RBMS1 in TCGA-HNSCC, GSE127165, GSE37991, and GSE13601 databases. B Western blot detection of RBMS1 protein expression in tissues (representative image); C qRT-PCR analysis of RBMS1 mRNA expression in 20 paired HNSCC samples ( n = 20, ****, p < 0.0001); D IHC staining to detect the expression and localization of RBMS1 in HNSCC specimens. Scale = 40 μm

Article Snippet: The blocks were cut into 4 μm thick sections and stained with Anti-Ki67 antibody (ab15580, Abcam,1/200), Anti-RBMS1 antibody (MA5-27,025, ThermoFisher, 1:150 dilution), and Anti-HSPA8 (1:200 dilution).

Techniques: Expressing, Biomarker Discovery, Western Blot, Quantitative RT-PCR, Immunohistochemistry

Journal: BMC Cancer

Article Title: RBMS1-HSPA8 axis activation drives head and neck squamous cell carcinoma progression

doi: 10.1186/s12885-025-13937-z

Figure Lengend Snippet: Effects of RBMS1 Knockdown and Overexpression on HNSCC Cell Phenotypes. A - B qRT-PCR and Western blot analysis of RBMS1 expression in NOK and three HNSCC cell lines. C - D Western blot and qRT-PCR validation of RBMS1 knockdown and overexpression efficiency in TU177 and FaDu cells (**** p < 0.0001). E – H CCK-8 and colony formation assays demonstrate that RBMS1 knockdown in TU177 cells reduces proliferation and colony formation, while RBMS1 overexpression in FaDu cells enhances these phenotypes compared to controls (shsc or Lv). I - J Wound healing assays show that RBMS1 knockdown in TU177 cells slows migration, whereas RBMS1 overexpression in FaDu cells accelerates wound closure. K - L Transwell assays reveal that RBMS1 knockdown in TU177 cells reduces migration and invasion, while RBMS1 overexpression in FaDu cells significantly enhances these capabilities

Article Snippet: The blocks were cut into 4 μm thick sections and stained with Anti-Ki67 antibody (ab15580, Abcam,1/200), Anti-RBMS1 antibody (MA5-27,025, ThermoFisher, 1:150 dilution), and Anti-HSPA8 (1:200 dilution).

Techniques: Knockdown, Over Expression, Quantitative RT-PCR, Western Blot, Expressing, Biomarker Discovery, CCK-8 Assay, Migration

Journal: BMC Cancer

Article Title: RBMS1-HSPA8 axis activation drives head and neck squamous cell carcinoma progression

doi: 10.1186/s12885-025-13937-z

Figure Lengend Snippet: RBMS1 promotes HNSCC tumor growth in vivo. Cells were subcutaneously injected into nude mice, and tumor growth was monitored. Tumor images ( A , E ), volume at specified time points ( B , F ), weight ( C , G ), and representative IHC staining of RBMS1 and Ki-67 are shown ( D , H ). Error bars represent the mean ± standard deviation (n = 3 per group). Scale bar = 50 μm

Article Snippet: The blocks were cut into 4 μm thick sections and stained with Anti-Ki67 antibody (ab15580, Abcam,1/200), Anti-RBMS1 antibody (MA5-27,025, ThermoFisher, 1:150 dilution), and Anti-HSPA8 (1:200 dilution).

Techniques: In Vivo, Injection, Immunohistochemistry, Standard Deviation

Journal: BMC Cancer

Article Title: RBMS1-HSPA8 axis activation drives head and neck squamous cell carcinoma progression

doi: 10.1186/s12885-025-13937-z

Figure Lengend Snippet: RBMS1 regulates HSPA8 expression. A Display 1314 differential genes using volcano map. B - D GO analysis of differentially expressed genes. E shRBMS1 and shSc TU177 cells. F RBMS1-overexpressing FaDu and the control cells. E – F HSPA8 levels were examined by Western blot (left panels) and qRT-PCR (right panels). G shRBMS1 TU177 cells were infected with lentiviruses carrying an empty vector (vector) or expression vectors for HSPA8. H RBMS1-overexpressing FaDu cells were infected with lentivirus harboring HSPA8 shRNA or a scrambled shRNA (shSc). RBMS1, HSPA8 protein and mRNA levels were examined by Western blot and qRT-PCR

Article Snippet: The blocks were cut into 4 μm thick sections and stained with Anti-Ki67 antibody (ab15580, Abcam,1/200), Anti-RBMS1 antibody (MA5-27,025, ThermoFisher, 1:150 dilution), and Anti-HSPA8 (1:200 dilution).

Techniques: Expressing, Control, Western Blot, Quantitative RT-PCR, Infection, Plasmid Preparation, shRNA

Journal: BMC Cancer

Article Title: RBMS1-HSPA8 axis activation drives head and neck squamous cell carcinoma progression

doi: 10.1186/s12885-025-13937-z

Figure Lengend Snippet: HSPA8 is a functionally important downstream target of RBMS1 in HNSCC cells. A - C , G - H , K - L . shRBMS1-1 TU177 cells were infected with lentiviruses carrying an empty vector or expression vectors for HSPA8. D - F , I - J , M – N RBMS1-overexpressing FaDu cells were infected with lentivirus harboring HSPA8 shRNAs or a scrambled shRNA. The cell proliferation and migratory abilities of the indicated cells were evaluated by CCK-8 ( A and D ), colony formation assays ( B - C and E – F ), and wound healing assays ( G - H and I - J ). K - L and M – N Cell migration and invasion abilities of the indicated cells were measured by transwell assays

Article Snippet: The blocks were cut into 4 μm thick sections and stained with Anti-Ki67 antibody (ab15580, Abcam,1/200), Anti-RBMS1 antibody (MA5-27,025, ThermoFisher, 1:150 dilution), and Anti-HSPA8 (1:200 dilution).

Techniques: Infection, Plasmid Preparation, Expressing, shRNA, CCK-8 Assay, Migration

Journal: BMC Cancer

Article Title: RBMS1-HSPA8 axis activation drives head and neck squamous cell carcinoma progression

doi: 10.1186/s12885-025-13937-z

Figure Lengend Snippet: RBMS1 promotes tumor growth through HSPA8 upregulation. Tumor growth of mice subcutaneously inoculated with the indicated cells. N = 3 for each group. A and E Tumor pictures. B and F Tumor growth curves. C and G Tumor weight. D and H Representative IHC staining for HSPA8 and Ki-67 of the indicated tumor tissues

Article Snippet: The blocks were cut into 4 μm thick sections and stained with Anti-Ki67 antibody (ab15580, Abcam,1/200), Anti-RBMS1 antibody (MA5-27,025, ThermoFisher, 1:150 dilution), and Anti-HSPA8 (1:200 dilution).

Techniques: Immunohistochemistry

Journal: Journal of Translational Medicine

Article Title: m6A methylation-modified ETV1 drives multiple myeloma progression and M2 polarization of tumor-associated macrophage through transcriptional activation of RBMS1

doi: 10.1186/s12967-026-07799-7

Figure Lengend Snippet: ETV1 activates RBMS1 transcription. ( A - B ) The expression of human RBMS1 in mouse bone marrow tissues and MM cells was verified by qPCR and western blot. ( C ) The luciferase reporter vectors containing the full-length or truncated promoter sequences of RBMS1 were inserted into pGL3-Basic plasmid and co-transfected with the ETV1 overexpression plasmid into RPMI8226 cells. After 48 h, the luciferase activity was measured by the kit. ( D ) The binding of exogenous ETV1 and RBMS1 promoter was detected by ChIP-PCR. Data are mean ± SD. ##, p < 0.01; ###, p < 0.001; ####, p < 0.0001

Article Snippet: The membranes were then incubated overnight at 4 °C with the following primary antibodies: human ETV1 (1: 1000 dilution, AP51197, Abcepta), human cyclinD1 (1: 5000 dilution, 26939-1-AP, Proteintech, Wuhan, China), human RBMS1 (1: 1000 dilution, 11061-2-AP, Proteintech) and human CCL2 (1: 500 dilution, 507277, Zen-bioscience, Chengdu, China).

Techniques: Expressing, Western Blot, Luciferase, Plasmid Preparation, Transfection, Over Expression, Activity Assay, Binding Assay

Journal: Journal of Translational Medicine

Article Title: m6A methylation-modified ETV1 drives multiple myeloma progression and M2 polarization of tumor-associated macrophage through transcriptional activation of RBMS1

doi: 10.1186/s12967-026-07799-7

Figure Lengend Snippet: RBMS1 knockdown counteracts the effect of ETV1 overexpression on MM cell proliferation and M2 polarization of TAMs. ( A ) siRNAs targeting RBMS1 were transfected into RPMI8226 cells. After 48 h, the knockdown efficiency of RBMS1 in cells was verified by qPCR. ( B ) Cell viability was measured by CCK-8 assay. ( C ) Cell cycle distribution was detected by flow cytometry. ( D ) The expression of cyclinD1 in cells was detected by western blot. ( E ) The levels of CCL2 in the cell supernatant were determined by ELISA. ( F ) M2 polarization of TAMs was tested by Transwell co-culture system. M0 macrophages were seeded into the lower chamber, and MM cells were seeded into the upper chamber. After 48 h of co-culture, the expression of CD163 and CD206 in M0 macrophages were determined by qPCR. ( G ) The proportion of CD206 + cells and MFI in M0 macrophages were examined by flow cytometry. Data are mean ± SD. #, p < 0.05; ##, p < 0.01; ###, p < 0.001; ####, p < 0.0001

Article Snippet: The membranes were then incubated overnight at 4 °C with the following primary antibodies: human ETV1 (1: 1000 dilution, AP51197, Abcepta), human cyclinD1 (1: 5000 dilution, 26939-1-AP, Proteintech, Wuhan, China), human RBMS1 (1: 1000 dilution, 11061-2-AP, Proteintech) and human CCL2 (1: 500 dilution, 507277, Zen-bioscience, Chengdu, China).

Techniques: Knockdown, Over Expression, Transfection, CCK-8 Assay, Flow Cytometry, Expressing, Western Blot, Enzyme-linked Immunosorbent Assay, Co-Culture Assay

Journal: Journal of Translational Medicine

Article Title: m6A methylation-modified ETV1 drives multiple myeloma progression and M2 polarization of tumor-associated macrophage through transcriptional activation of RBMS1

doi: 10.1186/s12967-026-07799-7

Figure Lengend Snippet: Schematic diagram demonstrating the mechanism of ETV1 and RBMS1 to promote MM progression and M2 polarization of TAMs

Article Snippet: The membranes were then incubated overnight at 4 °C with the following primary antibodies: human ETV1 (1: 1000 dilution, AP51197, Abcepta), human cyclinD1 (1: 5000 dilution, 26939-1-AP, Proteintech, Wuhan, China), human RBMS1 (1: 1000 dilution, 11061-2-AP, Proteintech) and human CCL2 (1: 500 dilution, 507277, Zen-bioscience, Chengdu, China).

Techniques:

Journal: Journal of Translational Medicine

Article Title: m6A methylation-modified ETV1 drives multiple myeloma progression and M2 polarization of tumor-associated macrophage through transcriptional activation of RBMS1

doi: 10.1186/s12967-026-07799-7

Figure Lengend Snippet: MM cell-derived ETV1 fosters M2 polarization of TAMs. ( A ) THP-1 cells were incubated with 350 nM PMA for 24 h to induce M0 macrophage differentiation. The morphology of M0 macrophages was photographed with a microscope. ( B ) The proportion of CD68 + cells in M0 macrophages were detected by flow cytometry. ( C ) M2 polarization of TAMs was measured by Transwell co-culture system. M0 macrophages were seeded into the lower chamber, and MM cells were seeded into the upper chamber. ( D ) The levels of CCL2 in MM cells was tested by qPCR and ELISA. ( E ) After 48 h of co-culture, the expression of CD163, Arg-1 and CD206 in M0 macrophages was determined by qPCR. ( F ) The proportion of CD206 + cells and MFI in M0 macrophages were examined by flow cytometry. Data are mean ± SD. #, p < 0.05; ##, p < 0.01; ###, p < 0.001; ####, p < 0.0001

Article Snippet: The membranes were then incubated overnight at 4 °C with the following primary antibodies: human ETV1 (1: 1000 dilution, AP51197, Abcepta), human cyclinD1 (1: 5000 dilution, 26939-1-AP, Proteintech, Wuhan, China), human RBMS1 (1: 1000 dilution, 11061-2-AP, Proteintech) and human CCL2 (1: 500 dilution, 507277, Zen-bioscience, Chengdu, China).

Techniques: Derivative Assay, Incubation, Microscopy, Flow Cytometry, Co-Culture Assay, Enzyme-linked Immunosorbent Assay, Expressing

Journal: Journal of Translational Medicine

Article Title: m6A methylation-modified ETV1 drives multiple myeloma progression and M2 polarization of tumor-associated macrophage through transcriptional activation of RBMS1

doi: 10.1186/s12967-026-07799-7

Figure Lengend Snippet: ETV1 facilitates tumor growth and M2 polarization of macrophages in vivo. ( A ) The development of tumors was determined by the in vivo fluorescence imaging system. ( B ) H&E staining was used to detect tumor formation in bone tissues of mice. (Scale bar = 100 μm). ( C ) Immunohistochemical staining was used to examine the expression of human ETV1 in bone marrow tissues. (Scale bar = 50 μm). ( D ) Flow cytometry was used to test the proportion of mouse-derived M2 phenotypic macrophages (F4/80 + CD206+/F4/80+) in M0 macrophages (F4/80+) in bone marrow tissues. ( E ) The expression of human CCL2 in bone marrow tissues was detected by western blot. Data are mean ± SD. ##, p < 0.01; ###, p < 0.001; ####, p < 0.0001

Article Snippet: The membranes were then incubated overnight at 4 °C with the following primary antibodies: human ETV1 (1: 1000 dilution, AP51197, Abcepta), human cyclinD1 (1: 5000 dilution, 26939-1-AP, Proteintech, Wuhan, China), human RBMS1 (1: 1000 dilution, 11061-2-AP, Proteintech) and human CCL2 (1: 500 dilution, 507277, Zen-bioscience, Chengdu, China).

Techniques: In Vivo, Fluorescence, Imaging, Staining, Immunohistochemical staining, Expressing, Flow Cytometry, Derivative Assay, Western Blot

Journal: Journal of Translational Medicine

Article Title: m6A methylation-modified ETV1 drives multiple myeloma progression and M2 polarization of tumor-associated macrophage through transcriptional activation of RBMS1

doi: 10.1186/s12967-026-07799-7

Figure Lengend Snippet: RBMS1 knockdown counteracts the effect of ETV1 overexpression on MM cell proliferation and M2 polarization of TAMs. ( A ) siRNAs targeting RBMS1 were transfected into RPMI8226 cells. After 48 h, the knockdown efficiency of RBMS1 in cells was verified by qPCR. ( B ) Cell viability was measured by CCK-8 assay. ( C ) Cell cycle distribution was detected by flow cytometry. ( D ) The expression of cyclinD1 in cells was detected by western blot. ( E ) The levels of CCL2 in the cell supernatant were determined by ELISA. ( F ) M2 polarization of TAMs was tested by Transwell co-culture system. M0 macrophages were seeded into the lower chamber, and MM cells were seeded into the upper chamber. After 48 h of co-culture, the expression of CD163 and CD206 in M0 macrophages were determined by qPCR. ( G ) The proportion of CD206 + cells and MFI in M0 macrophages were examined by flow cytometry. Data are mean ± SD. #, p < 0.05; ##, p < 0.01; ###, p < 0.001; ####, p < 0.0001

Article Snippet: The membranes were then incubated overnight at 4 °C with the following primary antibodies: human ETV1 (1: 1000 dilution, AP51197, Abcepta), human cyclinD1 (1: 5000 dilution, 26939-1-AP, Proteintech, Wuhan, China), human RBMS1 (1: 1000 dilution, 11061-2-AP, Proteintech) and human CCL2 (1: 500 dilution, 507277, Zen-bioscience, Chengdu, China).

Techniques: Knockdown, Over Expression, Transfection, CCK-8 Assay, Flow Cytometry, Expressing, Western Blot, Enzyme-linked Immunosorbent Assay, Co-Culture Assay

Journal: The American Journal of Pathology

Article Title: Identification of a Growth-Promoting Gene Cluster in the Region 2q24 as a Driver of Tumorigenesis in Childhood Hepatoblastoma

doi: 10.1016/j.ajpath.2025.04.006

Figure Lengend Snippet: A and B: Relative RNA expression levels of RBMS1 , BAZ2B , MARCH7 , DPP4 , FIGN , and TANK ( A ) and MYCN and MYC ( B ) are presented. A-1 and B-1: GeneChip human Transcriptome Array 2.0 (HTA2) was used on pediatric mature liver ( n = 6; orange), fetal liver ( n = 7; brown), pediatric hepatocellular carcinoma (HCC; n = 9; yellow), used HB cell lines ( n = 3; black), and fresh-frozen HB ( n 1 = 30; blue tones) cases; the y axis represents the log 10 scale of gene expression normalized to the mean sample signal. A-2 and B-2: Nanostring method was used on formalin-fixed, paraffin-embedded (FFPE) HB ( n 2 = 35; blue tones) cases; the y axis represents the log 2 scale of gene expression normalized to the expression of the housekeeping gene PNN . On the basis of molecular inversion probe data, fresh-frozen and FFPE HB samples were assigned as 2q stable ( n 1 = 12/ n 2 = 12; light blue), as (whole) 2q gain ( n 1 = 13/ n 2 = 19; medium blue), or as 2q24 high-gain/amplification (hg/amp; n 1 = 5/ n 2 = 4; dark blue). A-1,2 and B-1,2: The x axis shows the color code that represents the individual groups mentioned above. A-1 and B-1: Expression values of 2q24-amplified HB cases are highlighted in red triangles. All results are presented using box plots showing all points of each sample measurement. Bars with asterisks indicate significant group differences assumed at 95% confidence level. For simplicity, only the 2q24 high-gain/amplification group differences are presented. ∗ P < 0.05, ∗∗ P < 0.01, ∗∗∗ P < 0.001, and ∗∗∗∗ P < 0.0001.

Article Snippet: Sections were stained by IHC with antibodies against RBMS1 (mouse monoclonal antibody, clone OTI2H1, number TA808580; OriGene Technologies, Rockville, MD), MARCHF7 (rabbit polyclonal antibody, number NBP1-90056; RRID: AB_11044732 ; Novus Biologicals, Centennial, CO), DPP4 (goat polyclonal antibody, number AF1180; RRID: AB_354651 ; R&D Systems, Minneapolis, MN), FIGN (rabbit polyclonal antibody, number LS-C498842; LifeSpan BioSciences, Seattle, WA), and TANK (rabbit polyclonal antibody, number ABIN957231; antibodies-online, Limerick, PA).

Techniques: RNA Expression, Gene Expression, Formalin-fixed Paraffin-Embedded, Expressing, Amplification

Journal: The American Journal of Pathology

Article Title: Identification of a Growth-Promoting Gene Cluster in the Region 2q24 as a Driver of Tumorigenesis in Childhood Hepatoblastoma

doi: 10.1016/j.ajpath.2025.04.006

Figure Lengend Snippet: Expression levels and histologic distribution of the proteins RNA-binding motif, single-stranded-interacting protein 1 (RBMS1), E3 ubiquitin-protein ligase MARCHF7 (MARCH7), dipeptidyl peptidase 4 (DPP4), fidgetin (FIGN), and TRAF family member-associated NF-kappa-B activator (TANK). A – C: Histologic ( A ) and immunohistochemical ( B and C ) staining of pediatric mature liver, 2q stable HB (HB-75), 2q gain HB (HB-76), and 2q24 high-gain HB (HB-74) tissue sections. B: For comparison, the corresponding mRNA expression levels of the selected genes in these four cases (pediatric mature liver, HB-75, HB-76, and HB-74) are additionally presented. A: Histologic staining was conducted using hematoxylin and eosin (H&E) to evaluate histology (HB-75, fetal; HB-76, mixed epithelial-mesenchymal; and HB-74, mixed epithelial-mesenchymal). B and C: Immunohistochemistry staining was performed with specific antibodies against selected genes ( RBMS1 , MARCH7 , DPP4 , FIGN , and TANK ; B ) and β-catenin ( C ). B: Highest staining of RBMS1, MARCH7, DPP4, FIGN, and TANK was found in HB-74 (2q24 high-gain), which correlated with the corresponding mRNA expression levels. C: In HB-74 (2q24 high-gain), β-catenin showed overexpression with a cytosolic and nuclear shift, indicating activation of the Wnt pathway. The presented mRNA expression levels were analyzed by Nanostring method; the y axis represents the log 2 scale of gene expression normalized to the expression of the housekeeping gene PNN . Scale bars: 200 μm ( main images ); 15 μm ( insets ). Original magnification: ×20 ( main images ); ×80 ( insets ).

Article Snippet: Sections were stained by IHC with antibodies against RBMS1 (mouse monoclonal antibody, clone OTI2H1, number TA808580; OriGene Technologies, Rockville, MD), MARCHF7 (rabbit polyclonal antibody, number NBP1-90056; RRID: AB_11044732 ; Novus Biologicals, Centennial, CO), DPP4 (goat polyclonal antibody, number AF1180; RRID: AB_354651 ; R&D Systems, Minneapolis, MN), FIGN (rabbit polyclonal antibody, number LS-C498842; LifeSpan BioSciences, Seattle, WA), and TANK (rabbit polyclonal antibody, number ABIN957231; antibodies-online, Limerick, PA).

Techniques: Expressing, RNA Binding Assay, Ubiquitin Proteomics, Immunohistochemical staining, Staining, Comparison, Immunohistochemistry, Over Expression, Activation Assay, Gene Expression

Journal: The American Journal of Pathology

Article Title: Identification of a Growth-Promoting Gene Cluster in the Region 2q24 as a Driver of Tumorigenesis in Childhood Hepatoblastoma

doi: 10.1016/j.ajpath.2025.04.006

Figure Lengend Snippet: Impact of RBMS1 , BAZ2B , MARCH7 , DPP4 , FIGN , and TANK knockdown on proliferation rate and Wnt pathway activity in HB cell lines HepT1, HuH6, and HepG2. A: Immunoblot of RBMS1, c-Myc, and β-actin after RBMS1 knockdown (KD; cells after siRNA knockdown of RBMS1 ; Ctrl: cells transfected with scrambled control siRNA). β-Actin was used as a loading control. B: MTT assay for impact on cell proliferation rate. All bars show the mean of six replicates (light gray: cells transfected with scrambled control siRNA; dark gray: cells after siRNA knockdown of genes mentioned above). The y axis shows the proliferation rate of knockdown cells relative to scrambled control cells (set to 0) in percentage. C: Luciferase reporter assay for impact on Wnt pathway activity. All bars show the mean of six replicates (light gray: cells transfected with scrambled control siRNA; dark gray: cells after specific siRNA knockdown of selected genes). The y axis shows the Wnt pathway activity of knockdown cells relative to scrambled control cells (set to 0) in percentage. B and C: Error bars represent the SD. Bars with asterisks indicate significant group differences assumed at 95% confidence level. ∗ P < 0.05, ∗∗ P < 0.01, ∗∗∗ P < 0.001, and ∗∗∗∗ P < 0.0001.

Article Snippet: Sections were stained by IHC with antibodies against RBMS1 (mouse monoclonal antibody, clone OTI2H1, number TA808580; OriGene Technologies, Rockville, MD), MARCHF7 (rabbit polyclonal antibody, number NBP1-90056; RRID: AB_11044732 ; Novus Biologicals, Centennial, CO), DPP4 (goat polyclonal antibody, number AF1180; RRID: AB_354651 ; R&D Systems, Minneapolis, MN), FIGN (rabbit polyclonal antibody, number LS-C498842; LifeSpan BioSciences, Seattle, WA), and TANK (rabbit polyclonal antibody, number ABIN957231; antibodies-online, Limerick, PA).

Techniques: Knockdown, Activity Assay, Western Blot, Transfection, Control, MTT Assay, Luciferase, Reporter Assay