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

Journal: Cell Death & Disease

Article Title: SMYD3 drives the proliferation in gastric cancer cells via reducing EMP1 expression in an H4K20me3-dependent manner

doi: 10.1038/s41419-023-05907-9

Figure Lengend Snippet: A SMYD3 mRNA level is significantly upregulated in GC tissues ( n = 375) compared with normal tissues ( n = 32) from TCGA GC database ( p < 0.001). B SMYD3 mRNA expression is markedly increased in in-house GC tissues ( n = 30) by qPCR. C SMYD3 staining in GC tissues is stronger than that in normal tissues. Representative IHC images are shown here. A total of 125 pairs of tumor and normal tissues were analyzed. D SMYD3 staining was scored (0–12) and SMYD3 protein level was remarkably increased in GC samples relative to normal ones ( p < 0.001). E Higher SMYD3 expression is indicative of poorer overall survival rate from TCGA GC database ( p = 0.019). F Higher SMYD3 expression is indicative of poorer overall survival rate in institutional patients ( p = 0.005).

Article Snippet: The SMYD3 inhibitor BCI-121 was purchased from MedChemExpress (Princeton, NJ, USA) and dissolved in dimethyl sulfoxide (DMSO) at a stock concentration of 20 mM.

Techniques: Expressing, Staining

Journal: Cell Death & Disease

Article Title: SMYD3 drives the proliferation in gastric cancer cells via reducing EMP1 expression in an H4K20me3-dependent manner

doi: 10.1038/s41419-023-05907-9

Figure Lengend Snippet: Univariate and multivariate analysis.

Article Snippet: The SMYD3 inhibitor BCI-121 was purchased from MedChemExpress (Princeton, NJ, USA) and dissolved in dimethyl sulfoxide (DMSO) at a stock concentration of 20 mM.

Techniques: Expressing

Journal: Cell Death & Disease

Article Title: SMYD3 drives the proliferation in gastric cancer cells via reducing EMP1 expression in an H4K20me3-dependent manner

doi: 10.1038/s41419-023-05907-9

Figure Lengend Snippet: A SMYD3 expression in the immortalized human gastric epithelial cell line GES-1 and a set of GC cell lines were examined by western blotting assay. B The endogenous SMYD3 expression in HGC-27 and SGC-7901 cells was depleted using shRNAs verified by qPCR assay and western blotting assay. C SMYD3 depletion markedly inhibits the propagation in HGC-27 and SGC-7901 cells as shown by CCK8 assay. D SMYD3 depletion markedly suppresses the colony formation in HGC-27 and SGC-7901 cells. E SMYD3 depletion inhibited growth of tumors in vivo. F Tumor growth curves and tumor weight shows the suppressive effect of SMYD3 depletion in vivo. G IHC shows that Ki-67 staining was markedly weaker in tumor masses originated from SMYD3-depleted SGC-7901 cells. H The nuclear level of Geminin, Aurora A, p-Histone H3 (S10) and H4K20me3 dramatically decreased in HGC-27/SGC-7901 cells with SMYD3 knockdown.

Article Snippet: The SMYD3 inhibitor BCI-121 was purchased from MedChemExpress (Princeton, NJ, USA) and dissolved in dimethyl sulfoxide (DMSO) at a stock concentration of 20 mM.

Techniques: Expressing, Western Blot, CCK-8 Assay, In Vivo, Staining, Knockdown

Journal: Cell Death & Disease

Article Title: SMYD3 drives the proliferation in gastric cancer cells via reducing EMP1 expression in an H4K20me3-dependent manner

doi: 10.1038/s41419-023-05907-9

Figure Lengend Snippet: A The differentially expressed genes (DEGs) from RNA-seq data (GSE214155) were presented in the volcanic maps, with 345 genes altered in HGC-27 and 221 genes in SGC-7901 cells (|log2(FC)| > 0.5 and p < 0.05). The GC cohort from TCGA was stratified into the low (0–50%) and the high (50–100%) SMYD3-expression group, and the DEGs, with 10,954 altered between the two groups (|log2(FC)| > 0.5 and p < 0.05). Among the DEGs overlapped in the three datasets, EMP1 was the highest confidence target for SMYD3-H4K20me3. B Knockdown of endogenous SMYD3 expression elicited significantly increased EMP1 mRNA expression in HGC-27 and SGC-7901 cells by qPCR assay. C Knockdown of endogenous SMYD3 expression elicited significantly increased EMP1 protein expression in HGC-27 and SGC-7901 cells by immunoblot assay. D The specific SMYD3 DNA binding site (–CCCTCC-) P1/P2 in the EMP1 promoter. E ChIP assay revealed that SMYD3 and H4K20me3 were significantly enriched on the binding site P2. F EMP1 expression reversely correlated with SMYD3 expression in GC tissues ( r = −0.303, p < 0.001) in TCGA GC cohort. G SMYD3 mRNA level was reversely correlated with EMP1 expression in GES-1 cells and a panel of GC cell lines ( r = −0.621, p = 0.042) by qPCR assay. H SMYD3 protein level was reversely correlated with EMP1 expression in GES-1 cells and a panel of GC cell lines ( r = −0.621, p = 0.042) by immunoblot assay. I EMP1 protein level also correlated reversely with SMYD3 protein level ( r = −0.757, p = 0.011) via IHC in the same in-house GC specimens ( n = 125).

Article Snippet: The SMYD3 inhibitor BCI-121 was purchased from MedChemExpress (Princeton, NJ, USA) and dissolved in dimethyl sulfoxide (DMSO) at a stock concentration of 20 mM.

Techniques: RNA Sequencing Assay, Expressing, Knockdown, Western Blot, Binding Assay

Journal: Cell Death & Disease

Article Title: SMYD3 drives the proliferation in gastric cancer cells via reducing EMP1 expression in an H4K20me3-dependent manner

doi: 10.1038/s41419-023-05907-9

Figure Lengend Snippet: A KEGG pathway enrichment analysis of RNA-seq data (GSE214155) found that PI3K-Akt was the most influenced signaling pathway. B Immunoblotting analysis for H4K20me3 level, as well as EMP1, p-AKT(S473) and T-AKT in HGC-27 and SGC-7901 cells with SMYD3 deficiency. C Immunoblotting analysis for p-AKT(S473) level, as well as T-AKT in HGC-27 and SGC-7901 cells with exogenous expression of EMP1. D Immunoblotting analysis for H4K20me3 level, as well as EMP1, p-AKT(S473) and T-AKT in HGC-27 and SGC-7901 cells with SMYD3 deficiency combined with re-expression of SMYD3. E EMP1 expression was decreased in GC tissues ( N = 375) relative to normal ones ( N = 32) ( p < 0.05) in TCGA GC cohort. F EMP1 inhibits the cell growth in HGC-27 and SGC-7901 cells by CCK-8 assay. G EMP1 suppresses the colony formation in HGC-27 and SGC-7901 cells.

Article Snippet: The SMYD3 inhibitor BCI-121 was purchased from MedChemExpress (Princeton, NJ, USA) and dissolved in dimethyl sulfoxide (DMSO) at a stock concentration of 20 mM.

Techniques: RNA Sequencing Assay, Western Blot, Expressing, CCK-8 Assay

Journal: Cell Death & Disease

Article Title: SMYD3 drives the proliferation in gastric cancer cells via reducing EMP1 expression in an H4K20me3-dependent manner

doi: 10.1038/s41419-023-05907-9

Figure Lengend Snippet: A Reduced EMP1 expression relieved the deactivation of SMYD3 knockdown on the level of p-Akt (S473) in both HGC-27 and SGC-7901 cells by immunoblot assay. B Decreased EMP1 expression reignited the proliferation of HGC-27 and SGC-7901 cells in vitro by CCK-8 assay which was suppressed by knockdown of SMYD3 alone. C Decreased EMP1 expression reignited the proliferation of HGC-27 and SGC-7901 cells in vivo. D Tumor growth curves and tumor weight shows that decreased EMP1 expression reignited the GC cells proliferation in vivo.

Article Snippet: The SMYD3 inhibitor BCI-121 was purchased from MedChemExpress (Princeton, NJ, USA) and dissolved in dimethyl sulfoxide (DMSO) at a stock concentration of 20 mM.

Techniques: Expressing, Knockdown, Western Blot, In Vitro, CCK-8 Assay, In Vivo

Journal: Cell Death & Disease

Article Title: SMYD3 drives the proliferation in gastric cancer cells via reducing EMP1 expression in an H4K20me3-dependent manner

doi: 10.1038/s41419-023-05907-9

Figure Lengend Snippet: A SMYD3 inhibitor BCI-121 (100 μM) substantially mitigated the cellular proliferation of HGC-27 and SGC-7901 cells within the indicated time interval by CCK-8 assay. B BCI-121 remarkably reduced the level of H4K20me3 and p-Akt (S473), as well as increased EMP1 protein level in HGC-27 and SGC-7901 cells by immunoblot assay. C Intratumoral injection of BCI-121 (100 μM) markedly repressed the growth of SGC-7901 cells in vivo. D Tumor growth curves and tumor weight shows that BCI-121 markedly repressed the growth of SGC-7901 cells in vivo. E IHC shows that Ki-67 staining was markedly weaker in tumor masses originated from BCI-121-treated mice. F Administration of BCI-121 in vivo did not elicit evident damages to the livers and kidneys of the BCI-121-treated mice by HE staining assay. G Schematic diagram: SMYD3 promoted Akt signaling pathway and thus the proliferation via H4K20me3-mediated suppression of EMP1 expression in GC cells.

Article Snippet: The SMYD3 inhibitor BCI-121 was purchased from MedChemExpress (Princeton, NJ, USA) and dissolved in dimethyl sulfoxide (DMSO) at a stock concentration of 20 mM.

Techniques: CCK-8 Assay, Western Blot, Injection, In Vivo, Staining, Expressing

Journal: Cell Death & Disease

Article Title: SMYD3 drives the proliferation in gastric cancer cells via reducing EMP1 expression in an H4K20me3-dependent manner

doi: 10.1038/s41419-023-05907-9

Figure Lengend Snippet: Patient characteristics.

Article Snippet: The SMYD3 inhibitor BCI-121 was purchased from MedChemExpress (Princeton, NJ, USA) and dissolved in dimethyl sulfoxide (DMSO) at a stock concentration of 20 mM.

Techniques: Expressing

Journal: The Journal of Biological Chemistry

Article Title: The transcription factor HOXA9 induces expression of the chromatin modifier SMYD3 to drive leukemogenesis

doi: 10.1016/j.jbc.2025.110320

Figure Lengend Snippet: Combinational inhibition of SMYD3 and DOT1L induces enhanced growth arrest and differentiation in AML . A , synergistic effect of the combination of BCI-121 and EPZ5676 in MOLM-13 and THP-1 cells was analyzed by MTS assay after incubating cells with a serial diluted mixture at a fixed ratio of the two inhibitors. CI is plotted against the fraction effect. The reference line indicates CI = 1. CI values below 1 indicate synergism between the two inhibitors. B–D , co-treatment with BCI-121 and EPZ5676 led to enhanced inhibitory activity against the growth of AML cells. After being incubated with BCI-121 with or without EPZ5676 for 72 h, MOLM-13 and THP-1 cells were harvested, and then subjected to Western blotting analysis ( B ), dynamic cell counting ( C ), or colony formation ( D ), respectively. E– G , co-treatment with BCI-121 and EPZ5676 led to enhanced differentiation in AML cells. After exposed to BCI-121 with or without EPZ5676 for 72 h, MOLM-13 and THP-1 cells were subjected to Wright-Giemsa staining ( E ) and flow cytometry analysis for CD11b ( F ) or CD14 ( G ). Data are mean ± SD. ∗, p < 0.05; ∗∗, p < 0.01; ∗∗∗, p < 0.001; ∗∗∗∗, p < 0.0001, one-way ANOVA, Multiple comparison, Tukey's test was applied in C–G . All the experiments were repeated at least thrice.

Article Snippet: BCI-121 (T5322) and EPZ5676 (T3099) were purchased from TargetMol Chemicals (Shanghai, China).

Techniques: Inhibition, MTS Assay, Activity Assay, Incubation, Western Blot, Cell Counting, Staining, Flow Cytometry, Comparison

Journal: The Journal of Biological Chemistry

Article Title: The transcription factor HOXA9 induces expression of the chromatin modifier SMYD3 to drive leukemogenesis

doi: 10.1016/j.jbc.2025.110320

Figure Lengend Snippet: Combination of SMYD3 depletion and DOT1L inhibition induces enhanced growth arrest and differentiation in AML . A , MOLM-13 and THP-1 cells stably expressing Scramble or shSMYD3 were exposed to increasing concentrations of EPZ5676 for 72 h, and subjected to MTS assay. B–D , SMYD3 depletion sensitized AML cells to EPZ5676 treatment. After SMYD3 depletion, MOLM-13 and THP-1 cells were exposed to EPZ5676 for 72 h or not, and then subjected to Western blotting analysis ( B ), dynamic cell counting ( C ), or colony formation ( D ), respectively. E–G , SMYD3 depletion combined with EPZ5676 treatment led to enhanced differentiation in AML cells. After SMYD3 depletion, MOLM-13 and THP-1 cells were exposed to EPZ5676 for 72 h or not, and then subjected to Wright-Giemsa staining ( E ) or analyzed by flow cytometry for CD11b ( F ) or CD14 ( G ). H , a proposed model to describe the fundamental role of SMYD3 in AML. Data are mean ± SD. ∗, p < 0.05; ∗∗, p < 0.01; ∗∗∗, p < 0.001; ∗∗∗∗, p < 0.0001, one-way ANOVA, Multiple comparison, Tukey's test was applied in C–G . All the experiments were repeated at least thrice.

Article Snippet: BCI-121 (T5322) and EPZ5676 (T3099) were purchased from TargetMol Chemicals (Shanghai, China).

Techniques: Inhibition, Stable Transfection, Expressing, MTS Assay, Western Blot, Cell Counting, Staining, Flow Cytometry, Comparison