Journal: Cell Communication and Signaling : CCS

Article Title: SMYD3 synergises with RACK1 to promote colorectal cancer lung metastasis by recruiting SMAD3

doi: 10.1186/s12964-026-02687-5

Figure Lengend Snippet: SMYD3 is highly expressed in colorectal cancer tissues and correlates with poor prognosis. A Colorectal cancer mRNA expression profiling microarray analysis of differentially expressed genes with 154 common epimerase modifying enzymes taking the intersection Venn plot. B Heatmap of the expression profiles of 22 differentially expressed epimerase modifiers. C - D Western blot analysis of SMYD3 expression in colorectal cancer tissues (T) and corresponding paracancerous tissue (P) ( n = 8). E mRNA expression of SMYD3 in colorectal cancer tissues (T) and corresponding paracancerous tissue (P) ( n = 8). F Representative immunohistochemical (IHC) staining of SMYD3 expression in colorectal tumors and corresponding paracancerous tissue.( n = 96). G Immunohistochemical (IHC) scoring of SMYD3 expression in colorectal tumors and corresponding paracancerous tissue ( n = 96). H Kaplan–Meier curves of SMYD3 expression levels in tumour and normal tissues and patient prognosis(high: n = 62, low: n = 34). For D and F, significance was determined with the student unpaired t test.For G, significance was determined with Log–rank (Mantel–Cox) test. ns, not significant, p > 0.05; *, p < 0.05; **, p < 0.01;***, p < 0.001; ****, p < 0.0001. Errorbars, ± SD

Article Snippet: For the administration of the SMYD3 inhibitor BCI-121, nude mice injected with HCT116 mock and HCT116-shRACK1 cells were selected after tail vein injection and treated with BCI-121 (Selleck, #432,529–82-3) at a dose of 5 mg/kg via peritoneal injection (i.p.) three times per week for five weeks.

Techniques: Expressing, Microarray, Western Blot, Immunohistochemical staining, Immunohistochemistry

Journal: Cell Communication and Signaling : CCS

Article Title: SMYD3 synergises with RACK1 to promote colorectal cancer lung metastasis by recruiting SMAD3

doi: 10.1186/s12964-026-02687-5

Figure Lengend Snippet: SMYD3 promotes colorectal cancer cell metastasis. A Wound healing assays evaluating migration in CRC transfected with control or SMYD3 overexpression lentivirus and CRC mock cells, assessing migration. Scale bar = 50 μm. B - C Transwell experiment showing the migration capabilities of CRC cells transfected with control or SMYD3 overexpression lentivirus and CRC mock cells. Scale bar = 50 μm. D Wound-healing assays evaluating migration in CRC infected with control or SMYD3 knockdown lentivirus and CRC mock cells, assessing migration. Scale bar = 50 μm. E Transwell experiment showing the migration capabilities of CRC cells infected with control or SMYD3 knockdown lentivirus and CRC mock cells. Scale bar = 50 μm. F Statistical analysis of luciferase bioluminescence intensity ( n = 5). G Representative living images of mice injected with HCT116 infected by indicated lentivirus into tail vein. H Representative images of metastases in murine lung of each group; n = 5/group. The black arrow indicated the metastasis. I Statistical analysis of the number of pulmonary metastases of each group. J H&E staining of pulmonary tissue sections. For A,C,E,F and I, significance was determined with the student unpaired t test. ns, not significant, p > 0.05; *, p < 0.05; ***, p < 0.001; ****, p < 0.0001. Errorbars, ± SD

Article Snippet: For the administration of the SMYD3 inhibitor BCI-121, nude mice injected with HCT116 mock and HCT116-shRACK1 cells were selected after tail vein injection and treated with BCI-121 (Selleck, #432,529–82-3) at a dose of 5 mg/kg via peritoneal injection (i.p.) three times per week for five weeks.

Techniques: Migration, Transfection, Control, Over Expression, Infection, Knockdown, Luciferase, Injection, Staining

Journal: Cell Communication and Signaling : CCS

Article Title: SMYD3 synergises with RACK1 to promote colorectal cancer lung metastasis by recruiting SMAD3

doi: 10.1186/s12964-026-02687-5

Figure Lengend Snippet: SMYD3, RACK1 and SMAD3 interact with each other and the interaction between SMYD3 and SMAD3 depends on RACK1. A Silver staining results of proteins pulled down by Flag-SMYD3 in HCT116 cells. B The intersection veen plot of the immunoprecipitation and mass spectrometry analysis in colon cancer cell lines HCT15 and HCT116. C List of the candidate target proteins identified by the mass spectrometry. D Coimmunoprecipitation (Co-IP)-Western blot (WB) analysis of the SMYD3-RACK1 interaction in HCT116 cells transfected with Flag-tagged SMYD3 (Flag-SMYD3). E Co-IP-WB analysis of the SMYD3-RACK1-SMAD3 interaction in HCT116 cells transfected with Flag-tagged SMYD3 (Flag-SMYD3). F Immunoprecipitation experiments were performed to detect the endogenous interaction of SMYD3,SMAD3, RACK1 in HCT116 cells. G Immunofluorescence assay of endogenous SMYD3 and RACK1 in HCT116 cells. DAPI was used to counterstain the nucleus. H Immunofluorescence assay of endogenous SMYD3 and SMAD3 in HCT116 cells. DAPI was used to counterstain the nucleus. I Immunofluorescence assay of endogenous RACK1 and SMAD3 in HCT116 cells. DAPI was used to counterstain the nucleus. J Effects of RACK1 knockdown on SMYD3 interacted with SMAD3 in HCT116-LV-Flag-SMYD3 and RKO-LV-Flag-SMYD3 cells. K Effects of RACK1 knockdown on SMYD3 interacted with SMAD3 in HCT116-LV-His-SMAD3 and RKO-LV-His-SMAD3 cells

Article Snippet: For the administration of the SMYD3 inhibitor BCI-121, nude mice injected with HCT116 mock and HCT116-shRACK1 cells were selected after tail vein injection and treated with BCI-121 (Selleck, #432,529–82-3) at a dose of 5 mg/kg via peritoneal injection (i.p.) three times per week for five weeks.

Techniques: Silver Staining, Immunoprecipitation, Mass Spectrometry, Co-Immunoprecipitation Assay, Western Blot, Transfection, Immunofluorescence, Knockdown

Journal: Cell Communication and Signaling : CCS

Article Title: SMYD3 synergises with RACK1 to promote colorectal cancer lung metastasis by recruiting SMAD3

doi: 10.1186/s12964-026-02687-5

Figure Lengend Snippet: SMYD3 and SMAD3 co-regulate the expression of downstream TSKU. A Heatmap of putative SMYD3 direct target genes with upregulation or downregulation upon SMYD3 depletion. B Heatmap of putative SMAD3 direct target genes with upregulation or downregulation upon SMAD3 depletion. C Distribution of SMYD3 Cut-tag reads of binding signals around the 3-kb windows centeredon the transcription start site (TSS) of genes. D Pie-plot showing the distribution of SMYD3 in transcriptionally regulated regions of genes. E Venn diagram showing putative SMYD3 and SMAD3 direct target genes by combinational analyses of both RNA-seq, Cut-tag and ChIP-seq datasets. F Gene Ontology (GO) analysis of the top 10 biological processes enriched by the differentially expressed putative SMYD3 direct target genes. H RT–qPCR analysis of representative genes in HCT116 cells with SMYD3 or SMAD3 overexpression, individually. G Differential ploidy of target genes downstream of SMAD3-SMYD3. Data were shown as mean ± SD. The data were analyzed by Two-way ANOVA. *** p < 0.001

Article Snippet: For the administration of the SMYD3 inhibitor BCI-121, nude mice injected with HCT116 mock and HCT116-shRACK1 cells were selected after tail vein injection and treated with BCI-121 (Selleck, #432,529–82-3) at a dose of 5 mg/kg via peritoneal injection (i.p.) three times per week for five weeks.

Techniques: Expressing, Binding Assay, RNA Sequencing, ChIP-sequencing, Quantitative RT-PCR, Over Expression

Journal: Cell Communication and Signaling : CCS

Article Title: SMYD3 synergises with RACK1 to promote colorectal cancer lung metastasis by recruiting SMAD3

doi: 10.1186/s12964-026-02687-5

Figure Lengend Snippet: RACK1 recruits SMAD3 to SMYD3 and promotes the transcriptional activation of the genes downstream of SMYD3-SMAD3. A Effect of RACK1 knockdown on TSKU, H3K4me3, p-SMAD3 protein expression in RKO cells. B Effect of RACK1 overexpression on TSKU, H3K4me3, p-SMAD3 protein expression in HCT8 cells. C Effect of RACK1 knockdown on TSKU mRNA expression in RKO cells. D Effect of RACK1 overexpression on TSKU mRNA expression in HCT8 cells. E ChIP-qPCR with anti-Flag antibody showed binding of SMYD3 to TSKU. F ChIP-qPCR with anti-His antibody showed binding of SMAD3 to TSKU Data were shown as mean ± SD. The data were analyzed by Two-way ANOVA. *** p < 0.001

Article Snippet: For the administration of the SMYD3 inhibitor BCI-121, nude mice injected with HCT116 mock and HCT116-shRACK1 cells were selected after tail vein injection and treated with BCI-121 (Selleck, #432,529–82-3) at a dose of 5 mg/kg via peritoneal injection (i.p.) three times per week for five weeks.

Techniques: Activation Assay, Knockdown, Expressing, Over Expression, ChIP-qPCR, Binding Assay

Journal: Cell Communication and Signaling : CCS

Article Title: SMYD3 synergises with RACK1 to promote colorectal cancer lung metastasis by recruiting SMAD3

doi: 10.1186/s12964-026-02687-5

Figure Lengend Snippet: SMYD3-SMAD3 promotes colon cancer cell metastasis in vitro and in vivo dependent on RACK1. A Wound healing assay shows the effect of RACK1 knockdown on vector-, SMYD3-overexpressing and SMAD3-overexpressing RKO and HCT116 cell metastases. B Transwell assay shows the effect of RACK1 knockdown on vector-, SMYD3-overexpressing and SMAD3-overexpressing RKO and HCT116 cell metastases. C Wound healing assay shows the effect of RACK1 overexpression on vector-, SMYD3- knockdown and SMAD3- knockdown RKO and HCT8 cell metastases. D Transwell assay shows the effect of RACK1 overexpression on vector-, SMYD3- knockdown and SMAD3- knockdown RKO and HCT8 cell metastases. E The schematic diagram of tail vein lung metastasis model construction. F Representative living images of mice injected with HCT116 transfected by indicated lentivirus into tail vein. The lentivirus was Luci-labelled and therefore stably transfected HCT116 cell lines had in vivo luciferase activity. G Statistical analysis of luciferase bioluminescence intensity ( n = 5). H Statistical analysis of the number of pulmonary metastases of each group ( n = 5). I Representative images of metastases in murine lung of each group and H&E staining of pulmonary tissue sections; The black arrow indicated the metastasis. For A,B,C and D, significance was determined with the Two-way ANOVA. For G and H, significance was determined with the student unpaired t test. ns, not significant, p > 0.05; *, p < 0.05; ***, p < 0.001; ****, p < 0.0001. Errorbars, ± SD

Article Snippet: For the administration of the SMYD3 inhibitor BCI-121, nude mice injected with HCT116 mock and HCT116-shRACK1 cells were selected after tail vein injection and treated with BCI-121 (Selleck, #432,529–82-3) at a dose of 5 mg/kg via peritoneal injection (i.p.) three times per week for five weeks.

Techniques: In Vitro, In Vivo, Wound Healing Assay, Knockdown, Plasmid Preparation, Transwell Assay, Over Expression, Injection, Transfection, Stable Transfection, Luciferase, Activity Assay, Staining

Journal: Cell Communication and Signaling : CCS

Article Title: SMYD3 synergises with RACK1 to promote colorectal cancer lung metastasis by recruiting SMAD3

doi: 10.1186/s12964-026-02687-5

Figure Lengend Snippet: TSKU is highly expressed in colorectal cancer and associated with poor prognosis. A Box plot shows TSKU expression in Para-carcinoma and colorectal cancer tissues (Tumor) from TNMplot database. B mRNA expression of TSKU in colon cancer tissues (Tumor) and corresponding paracancerous tissue (P) ( n = 8). C Western blot analysis of TSKU expression in CRC tissues (T) and corresponding paracancerous tissue (P) ( n = 8). D Immunohistochemical (IHC) staining of TSKU expression levels in tumor and normal tissues.A total of 76 pairs of tumor and normal tissues were analysed. E Kaplan–Meier estimates of OS of patients with strong positive TSKU expression vs those with weak positive TSKU expression. F Representative images of immunohistochemical staining of SMYD3、SMAD3、 RACK1 and TSKU of colon cancer specimens ( n = 76). G TSKU expression correlates with SMYD3 levels in tissue microarray of colorectal cancer samples. Protein levels of TSKU and SMYD3 were quantified in colon cancer specimens. H TSKU expression correlates with SMAD3 levels in tissue microarray of colorectal cancer samples. Protein levels of TSKU and SMAD3 were quantified in colon cancer specimens. I TSKU expression correlates with RACK1 levels in tissue microarray of colorectal cancer samples. Protein levels of TSKU and RACK1 were quantified in colon cancer specimens. For B, significance was determined with the student unpaired t test. For E, significance was determined with Log–rank (Mantel–Cox) test. ns, not significant, p > 0.05; *, p < 0.05; ***, p < 0.001; ****, p < 0.0001. Errorbars, ± SD

Article Snippet: For the administration of the SMYD3 inhibitor BCI-121, nude mice injected with HCT116 mock and HCT116-shRACK1 cells were selected after tail vein injection and treated with BCI-121 (Selleck, #432,529–82-3) at a dose of 5 mg/kg via peritoneal injection (i.p.) three times per week for five weeks.

Techniques: Expressing, Western Blot, Immunohistochemical staining, Immunohistochemistry, Staining, Microarray

Journal: Cell Communication and Signaling : CCS

Article Title: SMYD3 synergises with RACK1 to promote colorectal cancer lung metastasis by recruiting SMAD3

doi: 10.1186/s12964-026-02687-5

Figure Lengend Snippet: SMYD3 promotes colorectal cancer cell metastasis. A Wound healing assays evaluating migration in CRC transfected with control or SMYD3 overexpression lentivirus and CRC mock cells, assessing migration. Scale bar = 50 μm. B - C Transwell experiment showing the migration capabilities of CRC cells transfected with control or SMYD3 overexpression lentivirus and CRC mock cells. Scale bar = 50 μm. D Wound-healing assays evaluating migration in CRC infected with control or SMYD3 knockdown lentivirus and CRC mock cells, assessing migration. Scale bar = 50 μm. E Transwell experiment showing the migration capabilities of CRC cells infected with control or SMYD3 knockdown lentivirus and CRC mock cells. Scale bar = 50 μm. F Statistical analysis of luciferase bioluminescence intensity ( n = 5). G Representative living images of mice injected with HCT116 infected by indicated lentivirus into tail vein. H Representative images of metastases in murine lung of each group; n = 5/group. The black arrow indicated the metastasis. I Statistical analysis of the number of pulmonary metastases of each group. J H&E staining of pulmonary tissue sections. For A,C,E,F and I, significance was determined with the student unpaired t test. ns, not significant, p > 0.05; *, p < 0.05; ***, p < 0.001; ****, p < 0.0001. Errorbars, ± SD

Article Snippet: For the administration of the SMYD3 inhibitor BCI-121, nude mice injected with HCT116 mock and HCT116-shRACK1 cells were selected after tail vein injection and treated with BCI-121 (Selleck, #432,529–82-3) at a dose of 5 mg/kg via peritoneal injection (i.p.) three times per week for five weeks.

Techniques: Migration, Transfection, Control, Over Expression, Infection, Knockdown, Luciferase, Injection, Staining

Journal: Cell Communication and Signaling : CCS

Article Title: SMYD3 synergises with RACK1 to promote colorectal cancer lung metastasis by recruiting SMAD3

doi: 10.1186/s12964-026-02687-5

Figure Lengend Snippet: SMYD3, RACK1 and SMAD3 interact with each other and the interaction between SMYD3 and SMAD3 depends on RACK1. A Silver staining results of proteins pulled down by Flag-SMYD3 in HCT116 cells. B The intersection veen plot of the immunoprecipitation and mass spectrometry analysis in colon cancer cell lines HCT15 and HCT116. C List of the candidate target proteins identified by the mass spectrometry. D Coimmunoprecipitation (Co-IP)-Western blot (WB) analysis of the SMYD3-RACK1 interaction in HCT116 cells transfected with Flag-tagged SMYD3 (Flag-SMYD3). E Co-IP-WB analysis of the SMYD3-RACK1-SMAD3 interaction in HCT116 cells transfected with Flag-tagged SMYD3 (Flag-SMYD3). F Immunoprecipitation experiments were performed to detect the endogenous interaction of SMYD3,SMAD3, RACK1 in HCT116 cells. G Immunofluorescence assay of endogenous SMYD3 and RACK1 in HCT116 cells. DAPI was used to counterstain the nucleus. H Immunofluorescence assay of endogenous SMYD3 and SMAD3 in HCT116 cells. DAPI was used to counterstain the nucleus. I Immunofluorescence assay of endogenous RACK1 and SMAD3 in HCT116 cells. DAPI was used to counterstain the nucleus. J Effects of RACK1 knockdown on SMYD3 interacted with SMAD3 in HCT116-LV-Flag-SMYD3 and RKO-LV-Flag-SMYD3 cells. K Effects of RACK1 knockdown on SMYD3 interacted with SMAD3 in HCT116-LV-His-SMAD3 and RKO-LV-His-SMAD3 cells

Article Snippet: For the administration of the SMYD3 inhibitor BCI-121, nude mice injected with HCT116 mock and HCT116-shRACK1 cells were selected after tail vein injection and treated with BCI-121 (Selleck, #432,529–82-3) at a dose of 5 mg/kg via peritoneal injection (i.p.) three times per week for five weeks.

Techniques: Silver Staining, Immunoprecipitation, Mass Spectrometry, Co-Immunoprecipitation Assay, Western Blot, Transfection, Immunofluorescence, Knockdown

Journal: Cell Communication and Signaling : CCS

Article Title: SMYD3 synergises with RACK1 to promote colorectal cancer lung metastasis by recruiting SMAD3

doi: 10.1186/s12964-026-02687-5

Figure Lengend Snippet: SMYD3 and SMAD3 co-regulate the expression of downstream TSKU. A Heatmap of putative SMYD3 direct target genes with upregulation or downregulation upon SMYD3 depletion. B Heatmap of putative SMAD3 direct target genes with upregulation or downregulation upon SMAD3 depletion. C Distribution of SMYD3 Cut-tag reads of binding signals around the 3-kb windows centeredon the transcription start site (TSS) of genes. D Pie-plot showing the distribution of SMYD3 in transcriptionally regulated regions of genes. E Venn diagram showing putative SMYD3 and SMAD3 direct target genes by combinational analyses of both RNA-seq, Cut-tag and ChIP-seq datasets. F Gene Ontology (GO) analysis of the top 10 biological processes enriched by the differentially expressed putative SMYD3 direct target genes. H RT–qPCR analysis of representative genes in HCT116 cells with SMYD3 or SMAD3 overexpression, individually. G Differential ploidy of target genes downstream of SMAD3-SMYD3. Data were shown as mean ± SD. The data were analyzed by Two-way ANOVA. *** p < 0.001

Article Snippet: For the administration of the SMYD3 inhibitor BCI-121, nude mice injected with HCT116 mock and HCT116-shRACK1 cells were selected after tail vein injection and treated with BCI-121 (Selleck, #432,529–82-3) at a dose of 5 mg/kg via peritoneal injection (i.p.) three times per week for five weeks.

Techniques: Expressing, Binding Assay, RNA Sequencing, ChIP-sequencing, Quantitative RT-PCR, Over Expression

Journal: Cell Communication and Signaling : CCS

Article Title: SMYD3 synergises with RACK1 to promote colorectal cancer lung metastasis by recruiting SMAD3

doi: 10.1186/s12964-026-02687-5

Figure Lengend Snippet: SMYD3-SMAD3 promotes colon cancer cell metastasis in vitro and in vivo dependent on RACK1. A Wound healing assay shows the effect of RACK1 knockdown on vector-, SMYD3-overexpressing and SMAD3-overexpressing RKO and HCT116 cell metastases. B Transwell assay shows the effect of RACK1 knockdown on vector-, SMYD3-overexpressing and SMAD3-overexpressing RKO and HCT116 cell metastases. C Wound healing assay shows the effect of RACK1 overexpression on vector-, SMYD3- knockdown and SMAD3- knockdown RKO and HCT8 cell metastases. D Transwell assay shows the effect of RACK1 overexpression on vector-, SMYD3- knockdown and SMAD3- knockdown RKO and HCT8 cell metastases. E The schematic diagram of tail vein lung metastasis model construction. F Representative living images of mice injected with HCT116 transfected by indicated lentivirus into tail vein. The lentivirus was Luci-labelled and therefore stably transfected HCT116 cell lines had in vivo luciferase activity. G Statistical analysis of luciferase bioluminescence intensity ( n = 5). H Statistical analysis of the number of pulmonary metastases of each group ( n = 5). I Representative images of metastases in murine lung of each group and H&E staining of pulmonary tissue sections; The black arrow indicated the metastasis. For A,B,C and D, significance was determined with the Two-way ANOVA. For G and H, significance was determined with the student unpaired t test. ns, not significant, p > 0.05; *, p < 0.05; ***, p < 0.001; ****, p < 0.0001. Errorbars, ± SD

Article Snippet: For the administration of the SMYD3 inhibitor BCI-121, nude mice injected with HCT116 mock and HCT116-shRACK1 cells were selected after tail vein injection and treated with BCI-121 (Selleck, #432,529–82-3) at a dose of 5 mg/kg via peritoneal injection (i.p.) three times per week for five weeks.

Techniques: In Vitro, In Vivo, Wound Healing Assay, Knockdown, Plasmid Preparation, Transwell Assay, Over Expression, Injection, Transfection, Stable Transfection, Luciferase, Activity Assay, Staining