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lentiviral shrna vectors targeting foxm1 pgv112 shfoxm1 (Genechem)

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Genechem lentiviral shrna vectors targeting foxm1 pgv112 shfoxm1

<t>FOXM1</t> promotes the proliferation of HCC cells in vitro . (A) The expression of FOXM1 in tumor tissues and adjacent tissues of patients with HCC analyzed by IHC. (B) The expression of Ki-67 in tumor tissues of patients with HCC analyzed by IHC. (C) The differential expression of FOXM1 between tumor tissues and adjacent tissues of patients with HCC analyzed by IHC. (D) Correlation analysis of FOXM1 and Ki-67 IHC scores in tumor tissues of patients with HCC. (E) Box plots for the differential expression of FOXM1 between LIHC tissues and adjacent tissues in TCGA. (F) ROC curves for the relationship between FOXM1 expression and the prognosis of HCC patients. (G) Correlation between FOXM1 expression levels and survival of HCC patients. (H) Distribution of FOXM1 mRNA expression in HCC cell lines. (I) Expression of FOXM1 in HUH7, HepG2, and SMMC-7721 cells analyzed by Western blot. (J) Effect of FOXM1 shRNA and FOXM1 cDNA on the viability of HUH7 cells. (K) Effects of FOXM1 on the formation of HUH7 colonies. (L) Effects of FOXM1 on the apoptosis of HUH7 cells. (M) Effects of FDI-6 on the proliferation of HUH7 cells. (N) Effects of FDI-6 on the formation of HUH7 colonies. (O) Effects of FOXM1 shRNA and FOXM1 cDNA on mouse weight. (P) Effects of FOXM1 shRNA and FOXM1 cDNA on tumor volume. (Q) The photos of tumor nodules in each group. (R) Effects of FOXM1 shRNA and FOXM1 cDNA on tumor weight. (S) Effects of FOXM1 shRNA and FOXM1 cDNA on tumor proliferation. (T) The expression of Ki-67 and FOXM1 in xenograft tumor analyzed by IHC. Images were randomly selected from five replicates. Data from three independent experiments were analyzed by one-way ANOVA: * P<0.05, ** P<0.01 vs. control group; # P<0.05, ## P<0.01 vs. FOXM1 KD group.

Lentiviral Shrna Vectors Targeting Foxm1 Pgv112 Shfoxm1, supplied by Genechem, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/foxm1-shrna/foxm1+lentiviral+pgv112+shfoxm1+shrna+targeting+vectors/pmc13050458-75-22-39
Average 86 stars, based on 1 article reviews

lentiviral shrna vectors targeting foxm1 pgv112 shfoxm1 - by Bioz Stars, 2026-09

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1) Product Images from "Mechanistic Insights into the FOXM1/BUB1 axis-Mediated Oncogenic Signaling in Hepatocellular Carcinoma"

Article Title: Mechanistic Insights into the FOXM1/BUB1 axis-Mediated Oncogenic Signaling in Hepatocellular Carcinoma

Journal: International Journal of Biological Sciences

doi: 10.7150/ijbs.125454

FOXM1 promotes the proliferation of HCC cells in vitro . (A) The expression of FOXM1 in tumor tissues and adjacent tissues of patients with HCC analyzed by IHC. (B) The expression of Ki-67 in tumor tissues of patients with HCC analyzed by IHC. (C) The differential expression of FOXM1 between tumor tissues and adjacent tissues of patients with HCC analyzed by IHC. (D) Correlation analysis of FOXM1 and Ki-67 IHC scores in tumor tissues of patients with HCC. (E) Box plots for the differential expression of FOXM1 between LIHC tissues and adjacent tissues in TCGA. (F) ROC curves for the relationship between FOXM1 expression and the prognosis of HCC patients. (G) Correlation between FOXM1 expression levels and survival of HCC patients. (H) Distribution of FOXM1 mRNA expression in HCC cell lines. (I) Expression of FOXM1 in HUH7, HepG2, and SMMC-7721 cells analyzed by Western blot. (J) Effect of FOXM1 shRNA and FOXM1 cDNA on the viability of HUH7 cells. (K) Effects of FOXM1 on the formation of HUH7 colonies. (L) Effects of FOXM1 on the apoptosis of HUH7 cells. (M) Effects of FDI-6 on the proliferation of HUH7 cells. (N) Effects of FDI-6 on the formation of HUH7 colonies. (O) Effects of FOXM1 shRNA and FOXM1 cDNA on mouse weight. (P) Effects of FOXM1 shRNA and FOXM1 cDNA on tumor volume. (Q) The photos of tumor nodules in each group. (R) Effects of FOXM1 shRNA and FOXM1 cDNA on tumor weight. (S) Effects of FOXM1 shRNA and FOXM1 cDNA on tumor proliferation. (T) The expression of Ki-67 and FOXM1 in xenograft tumor analyzed by IHC. Images were randomly selected from five replicates. Data from three independent experiments were analyzed by one-way ANOVA: * P<0.05, ** P<0.01 vs. control group; # P<0.05, ## P<0.01 vs. FOXM1 KD group.

Figure Legend Snippet: FOXM1 promotes the proliferation of HCC cells in vitro . (A) The expression of FOXM1 in tumor tissues and adjacent tissues of patients with HCC analyzed by IHC. (B) The expression of Ki-67 in tumor tissues of patients with HCC analyzed by IHC. (C) The differential expression of FOXM1 between tumor tissues and adjacent tissues of patients with HCC analyzed by IHC. (D) Correlation analysis of FOXM1 and Ki-67 IHC scores in tumor tissues of patients with HCC. (E) Box plots for the differential expression of FOXM1 between LIHC tissues and adjacent tissues in TCGA. (F) ROC curves for the relationship between FOXM1 expression and the prognosis of HCC patients. (G) Correlation between FOXM1 expression levels and survival of HCC patients. (H) Distribution of FOXM1 mRNA expression in HCC cell lines. (I) Expression of FOXM1 in HUH7, HepG2, and SMMC-7721 cells analyzed by Western blot. (J) Effect of FOXM1 shRNA and FOXM1 cDNA on the viability of HUH7 cells. (K) Effects of FOXM1 on the formation of HUH7 colonies. (L) Effects of FOXM1 on the apoptosis of HUH7 cells. (M) Effects of FDI-6 on the proliferation of HUH7 cells. (N) Effects of FDI-6 on the formation of HUH7 colonies. (O) Effects of FOXM1 shRNA and FOXM1 cDNA on mouse weight. (P) Effects of FOXM1 shRNA and FOXM1 cDNA on tumor volume. (Q) The photos of tumor nodules in each group. (R) Effects of FOXM1 shRNA and FOXM1 cDNA on tumor weight. (S) Effects of FOXM1 shRNA and FOXM1 cDNA on tumor proliferation. (T) The expression of Ki-67 and FOXM1 in xenograft tumor analyzed by IHC. Images were randomly selected from five replicates. Data from three independent experiments were analyzed by one-way ANOVA: * P<0.05, ** P<0.01 vs. control group; # P<0.05, ## P<0.01 vs. FOXM1 KD group.

Techniques Used: In Vitro, Expressing, Quantitative Proteomics, Western Blot, shRNA, Control

FOXM1-regulated genes involve in DNA repair, cell cycle, stemness and EMT. (A) GO function and KEGG pathway enrichment analysis of DEGs between FOXM1 KD and NC KD in HUH7 cells. (B) . GO function and KEGG pathway enrichment analysis of top 50 targets in the PPI network of FOXM1. (C) . Volcano plot for the DEGs involved in DNA repair and cell cycle analyzed by RNA-seq. (D) Volcano plot for the DEGs involved in cell stemness, invasion and migration. (E) . The relationship between FOXM1 expression and HCC cell stemness in TCGA. (F) . Differential expression of genes between LIHC tissues and adjacent tissues in TCGA. (G) . ROC curves for the relationship between gene expression and HCC prognosis. Data were statistically analyzed using one-way ANOVA: * P<0.05 compared with adjacent tissues.

Figure Legend Snippet: FOXM1-regulated genes involve in DNA repair, cell cycle, stemness and EMT. (A) GO function and KEGG pathway enrichment analysis of DEGs between FOXM1 KD and NC KD in HUH7 cells. (B) . GO function and KEGG pathway enrichment analysis of top 50 targets in the PPI network of FOXM1. (C) . Volcano plot for the DEGs involved in DNA repair and cell cycle analyzed by RNA-seq. (D) Volcano plot for the DEGs involved in cell stemness, invasion and migration. (E) . The relationship between FOXM1 expression and HCC cell stemness in TCGA. (F) . Differential expression of genes between LIHC tissues and adjacent tissues in TCGA. (G) . ROC curves for the relationship between gene expression and HCC prognosis. Data were statistically analyzed using one-way ANOVA: * P<0.05 compared with adjacent tissues.

Techniques Used: RNA Sequencing, Migration, Expressing, Quantitative Proteomics, Gene Expression

FOXM1 promotes DNA repair and G2/M progression on HCC cells. (A) Effects of FOXM1 on DNA damage in HUH7 and HepG2 cells analyzed by comet assay. (B) Effects of FOXM1 on γH2AX expression analyzed by IF. (C) Q-PCR analysis of the effects of FOXM1 on the expression of DNA repair-related genes. (D) Analysis of the correlation between FOXM1 and DNA repair-related gene expressions in LIHC. (E) Effects of FOXM1 on cell cycle progression in HUH7 cells. (F) percentage of cell cycle at G0/G1, S, and G2/M phases. (G) Q-PCR analysis of the effects of FOXM1 on the expression of genes regulating G2/M transition. (H) Analysis of the correlation between FOXM1 and cell cycle related gene expressions in HCC. (I) Effects of FDI-6 on DNA damage in HUH7 cells. (J) Effects of FDI-6 on the expression of DNA-repair related genes in HUH7 cells. (K) Effects of FDI-6 on cell cycle progression in HUH7 cells. (L) Effects of FDI-6 on cell cycle-related genes in HUH7 cells. Images were randomly selected from five replicates. Data from three independent experiments were statistically analyzed using one-way ANOVA: * P < 0.05, ** P < 0.01 vs. NC KD group; # P < 0.05, ## P < 0.01 vs. FOXM1 KD group.

Figure Legend Snippet: FOXM1 promotes DNA repair and G2/M progression on HCC cells. (A) Effects of FOXM1 on DNA damage in HUH7 and HepG2 cells analyzed by comet assay. (B) Effects of FOXM1 on γH2AX expression analyzed by IF. (C) Q-PCR analysis of the effects of FOXM1 on the expression of DNA repair-related genes. (D) Analysis of the correlation between FOXM1 and DNA repair-related gene expressions in LIHC. (E) Effects of FOXM1 on cell cycle progression in HUH7 cells. (F) percentage of cell cycle at G0/G1, S, and G2/M phases. (G) Q-PCR analysis of the effects of FOXM1 on the expression of genes regulating G2/M transition. (H) Analysis of the correlation between FOXM1 and cell cycle related gene expressions in HCC. (I) Effects of FDI-6 on DNA damage in HUH7 cells. (J) Effects of FDI-6 on the expression of DNA-repair related genes in HUH7 cells. (K) Effects of FDI-6 on cell cycle progression in HUH7 cells. (L) Effects of FDI-6 on cell cycle-related genes in HUH7 cells. Images were randomly selected from five replicates. Data from three independent experiments were statistically analyzed using one-way ANOVA: * P < 0.05, ** P < 0.01 vs. NC KD group; # P < 0.05, ## P < 0.01 vs. FOXM1 KD group.

Techniques Used: Single Cell Gel Electrophoresis, Expressing

FOXM1 drives cell stemness, invasion, and migration in HCC cells. (A) Effects of FOXM1 on CD44 expression analyzed by IF in HUH7 cells. (B) Effects of FOXM1 on CD44 expression in HepG2 cells. (C) Effects of FOXM1 on the expression of stemness-related genes in HUH7 cells. (D) Effects of FDI-6 on CD44 expression in HUH7 cells. (E) Effects of FDI-6 on the expression of stemness-related genes in HUH7 cells. (F) Effects of FOXM1 shRNA and FDI-6 on the formation of HUH7 three-dimensional spheres. (G) Analysis of the correlation between FOXM1 and cell cycle-related gene expressions in HCC. (H) Effects of FOXM1 on the invasion and migration of HUH7 and HepG2 cells. (I) Q-PCR analysis of the effects of FOXM1 on EMT-related gene expression in HUH7 cells. (J) Effects of FDI-6 on the invasion and migration of HUH7 cells. (K) Q-PCR analysis of the effects of FDI-6 on EMT-related gene expression in HUH7 cells. (L) Analysis of the correlation between FOXM1 and EMT-related gene expressions in HCC. Images were randomly selected from five replicates. Data from three independent experiments were statistically analyzed using one-way ANOVA: * P < 0.05, ** P < 0.01 vs. NC KD group; # P < 0.05, ## P < 0.01 vs. FOXM1 KD group.

Figure Legend Snippet: FOXM1 drives cell stemness, invasion, and migration in HCC cells. (A) Effects of FOXM1 on CD44 expression analyzed by IF in HUH7 cells. (B) Effects of FOXM1 on CD44 expression in HepG2 cells. (C) Effects of FOXM1 on the expression of stemness-related genes in HUH7 cells. (D) Effects of FDI-6 on CD44 expression in HUH7 cells. (E) Effects of FDI-6 on the expression of stemness-related genes in HUH7 cells. (F) Effects of FOXM1 shRNA and FDI-6 on the formation of HUH7 three-dimensional spheres. (G) Analysis of the correlation between FOXM1 and cell cycle-related gene expressions in HCC. (H) Effects of FOXM1 on the invasion and migration of HUH7 and HepG2 cells. (I) Q-PCR analysis of the effects of FOXM1 on EMT-related gene expression in HUH7 cells. (J) Effects of FDI-6 on the invasion and migration of HUH7 cells. (K) Q-PCR analysis of the effects of FDI-6 on EMT-related gene expression in HUH7 cells. (L) Analysis of the correlation between FOXM1 and EMT-related gene expressions in HCC. Images were randomly selected from five replicates. Data from three independent experiments were statistically analyzed using one-way ANOVA: * P < 0.05, ** P < 0.01 vs. NC KD group; # P < 0.05, ## P < 0.01 vs. FOXM1 KD group.

Techniques Used: Migration, Expressing, shRNA, Gene Expression

FOXM1 promotes BUB1 expression at transcriptional level. (A) Venn diagram of overlapping genes identified by RNA-seq and FOXM1 PPI network. (B) Analysis the association between key gene expression and HCC patient survival. (C) The expression of BUB1 between HCC tissues and adjacent tissues from clinical patients analyzed by IHC. (D) IHC scores of BUB1 between HCC tissues and adjacent tissues from clinical patients. (E) Correlation analysis of BUB1 and Ki-67 IHC scores in tumor tissues of patients with HCC. (F) Correlation analysis of BUB1 and FOXM1 IHC scores in tumor tissues of patients with HCC. (G) Co-IP analysis of the interaction between FOXM1 and BUB1 in HUH7 cells. (H) Effects of FOXM1 shRNA and BUB1 shRNA on the expression of FOXM1 and BUB1 analyzed by Q-PCR. (I) The expression of FOXM1 and BUB1 analyzed by Western blot. (J) ChIP-qPCR analysis of the binding of FOXM1 and BUB1 promoter in HUH7 cells. (K) The sequences at the -293 bp of the BUB1-WT promoter and the BUB1-Mut promoter. (L) The binding of FOXM1 and BUB1 promoter at the -293 bp sequence GTAAACC analyzed by dual luciferase reporter assay. (M) Q-PCR analysis of BUB1 expression in different HCC cells. (N) Western blot analysis of BUB1 expression in different HCC cells. (O) KEGG pathway enrichment analysis of top 50 targets in the PPI network of BUB1. (P) Venn diagram of overlapping genes identified by RNA-seq, FOXM1 PPI network, and BUB1 PPI network. (Q) correlation analysis of BUB1 expression and LIHC cell stemness in TCGA. (R) Effect of BUB1 on the inhibitory role of FOXM1 shRNA in the proliferation of HUH7 cells. (S) Effects of BUB1 on the inhibition of FOXM1 shRNA in HUH7 colony formation. (T) Effects of BUB1 on the promotion of FOXM1 shRNA in HUH7 cell apoptosis. (U) Effects of BUB1 shRNA on HUH7 xenograft tumor volumes. (V) Photos of HUH7 xenograft tumors in each group. (W) Effects of BUB1 shRNA on HUH7 xenograft tumor weight. (X) Effects of BUB1 shRNA on Ki-67 and BUB1 expression in each group. Images were randomly selected from five replicates. Data from three independent experiments were analyzed by one-way ANOVA: * P<0.05, ** P<0.01 vs. NC KD group, # P < 0.05, ## P < 0.01 vs. FOXM1 KD group.

Figure Legend Snippet: FOXM1 promotes BUB1 expression at transcriptional level. (A) Venn diagram of overlapping genes identified by RNA-seq and FOXM1 PPI network. (B) Analysis the association between key gene expression and HCC patient survival. (C) The expression of BUB1 between HCC tissues and adjacent tissues from clinical patients analyzed by IHC. (D) IHC scores of BUB1 between HCC tissues and adjacent tissues from clinical patients. (E) Correlation analysis of BUB1 and Ki-67 IHC scores in tumor tissues of patients with HCC. (F) Correlation analysis of BUB1 and FOXM1 IHC scores in tumor tissues of patients with HCC. (G) Co-IP analysis of the interaction between FOXM1 and BUB1 in HUH7 cells. (H) Effects of FOXM1 shRNA and BUB1 shRNA on the expression of FOXM1 and BUB1 analyzed by Q-PCR. (I) The expression of FOXM1 and BUB1 analyzed by Western blot. (J) ChIP-qPCR analysis of the binding of FOXM1 and BUB1 promoter in HUH7 cells. (K) The sequences at the -293 bp of the BUB1-WT promoter and the BUB1-Mut promoter. (L) The binding of FOXM1 and BUB1 promoter at the -293 bp sequence GTAAACC analyzed by dual luciferase reporter assay. (M) Q-PCR analysis of BUB1 expression in different HCC cells. (N) Western blot analysis of BUB1 expression in different HCC cells. (O) KEGG pathway enrichment analysis of top 50 targets in the PPI network of BUB1. (P) Venn diagram of overlapping genes identified by RNA-seq, FOXM1 PPI network, and BUB1 PPI network. (Q) correlation analysis of BUB1 expression and LIHC cell stemness in TCGA. (R) Effect of BUB1 on the inhibitory role of FOXM1 shRNA in the proliferation of HUH7 cells. (S) Effects of BUB1 on the inhibition of FOXM1 shRNA in HUH7 colony formation. (T) Effects of BUB1 on the promotion of FOXM1 shRNA in HUH7 cell apoptosis. (U) Effects of BUB1 shRNA on HUH7 xenograft tumor volumes. (V) Photos of HUH7 xenograft tumors in each group. (W) Effects of BUB1 shRNA on HUH7 xenograft tumor weight. (X) Effects of BUB1 shRNA on Ki-67 and BUB1 expression in each group. Images were randomly selected from five replicates. Data from three independent experiments were analyzed by one-way ANOVA: * P<0.05, ** P<0.01 vs. NC KD group, # P < 0.05, ## P < 0.01 vs. FOXM1 KD group.

Techniques Used: Expressing, RNA Sequencing, Gene Expression, Co-Immunoprecipitation Assay, shRNA, Western Blot, ChIP-qPCR, Binding Assay, Sequencing, Luciferase, Reporter Assay, Inhibition

Knockdown of BUB1 enhances HCC cell sensitivity to FOXM1 inhibitor FDI-6. (A) Effect of BUB1 shRNA on FDI-6-mediated inhibition of colony formation in HUH7 cells. (B) Effect of BUB1 shRNA on FDI-6-induced apoptosis. (C) Effect of BUB1 shRNA on FDI-6-induced DNA damage. (D) Effect of BUB1 shRNA on the G2/M phase arrest caused by FDI-6. (E) Western blot analysis of BUB1 shRNA and FDI-6 effects on DNA repair-related gene expression. (F) Western blot analysis of BUB1 shRNA and FDI-6 effects on cell cycle-related gene expression. (G) IF analysis of BUB1 shRNA and FDI-6 effects on CD44 expression. (H) Western blot analysis of BUB1 shRNA and FDI-6 effects on stemness-related gene expression. (I) Effects of BUB1 shRNA on FDI-6 mediated suppression of HCC cell invasion and migration. (J) Western blot analysis of BUB1 shRNA and FDI-6 effects on EMT-related gene expression. (K) Effects of BUB1 shRNA and FDI-6 on mouse weight. (L) Effects of BUB1 shRNA and FDI-6 on tumor volume. (M) The photos of HUH7 xenograft tumors in each group. (N) Effects of BUB1 shRNA and FDI-6 on tumor weight. (O) The inhibition ratios of BUB1 shRNA and FDI-6 on tumor volume and weight. (P) IHC analysis of the effects of BUB1 shRNA and FDI-6 on Ki-67 and FOXM1 expression Images were randomly selected from five replicates. Data from three independent experiments were statistically analyzed using one-way ANOVA: * P<0.05, ** P<0.01 vs. NC KD group, # P < 0.05, ## P < 0.01 vs. BUB1 KD+FDI-6 group.

Figure Legend Snippet: Knockdown of BUB1 enhances HCC cell sensitivity to FOXM1 inhibitor FDI-6. (A) Effect of BUB1 shRNA on FDI-6-mediated inhibition of colony formation in HUH7 cells. (B) Effect of BUB1 shRNA on FDI-6-induced apoptosis. (C) Effect of BUB1 shRNA on FDI-6-induced DNA damage. (D) Effect of BUB1 shRNA on the G2/M phase arrest caused by FDI-6. (E) Western blot analysis of BUB1 shRNA and FDI-6 effects on DNA repair-related gene expression. (F) Western blot analysis of BUB1 shRNA and FDI-6 effects on cell cycle-related gene expression. (G) IF analysis of BUB1 shRNA and FDI-6 effects on CD44 expression. (H) Western blot analysis of BUB1 shRNA and FDI-6 effects on stemness-related gene expression. (I) Effects of BUB1 shRNA on FDI-6 mediated suppression of HCC cell invasion and migration. (J) Western blot analysis of BUB1 shRNA and FDI-6 effects on EMT-related gene expression. (K) Effects of BUB1 shRNA and FDI-6 on mouse weight. (L) Effects of BUB1 shRNA and FDI-6 on tumor volume. (M) The photos of HUH7 xenograft tumors in each group. (N) Effects of BUB1 shRNA and FDI-6 on tumor weight. (O) The inhibition ratios of BUB1 shRNA and FDI-6 on tumor volume and weight. (P) IHC analysis of the effects of BUB1 shRNA and FDI-6 on Ki-67 and FOXM1 expression Images were randomly selected from five replicates. Data from three independent experiments were statistically analyzed using one-way ANOVA: * P<0.05, ** P<0.01 vs. NC KD group, # P < 0.05, ## P < 0.01 vs. BUB1 KD+FDI-6 group.

Techniques Used: Knockdown, shRNA, Inhibition, Western Blot, Gene Expression, Expressing, Migration

FOXM1 inhibitors and BAY synergistically inhibit proliferation of HCC cells and tumors. (A) Effect of BAY and FDI-6 on the proliferation of HUH7 cells. (B) The CI values of the combinations of BAY and FDI-6 at different concentration ratios. (C) Effect of BAY, FDI-6 and their combination on the colony formation of HUH7 cells. (D) Effect of BAY, FDI-6 and their combination on the apoptosis of HUH7 cells. (E) The CI values of the combinations of TST and FDI-6 at different concentration ratios. (F) Effect of TST, FDI-6 and their combination on the colony formation of HUH7 cells. (G) The CI values of the combinations of RCM-1 and FDI-6 at different concentration ratios. (H) Effect of RCM-1, FDI-6 and their combination on the colony formation of HUH7 cells. (I) Effect of BAY, FDI-6 and their sequential combination on tumor volume. (J) Effect of BAY, FDI-6 and their sequential combination on tumor weight. (K) The photos of HUH7 xenograft tumors in each group. (L) Acute toxicity analysis of 100 mg/kg FDI-6, 100 mg/kg BAY, and their combination in mice. (M) Effect of BAY, FDI-6 and their sequential combination on mouse weight. (N) Effect of BAY, FDI-6 and their sequential combination on the heart, liver, spleen, lung and kidney in mice. (O) Effect of BAY, FDI-6 and their sequential combination on the expression of Ki-67, FOXM1, and BUB1 in HUH7 xenograft tumors. Images were randomly selected from five replicates. Data from three independent experiments were statistically analyzed using one-way ANOVA: * P<0.05, ** P<0.01 vs. control group, # P < 0.05, ## P < 0.01 vs. BAY-FDI-6 group.

Figure Legend Snippet: FOXM1 inhibitors and BAY synergistically inhibit proliferation of HCC cells and tumors. (A) Effect of BAY and FDI-6 on the proliferation of HUH7 cells. (B) The CI values of the combinations of BAY and FDI-6 at different concentration ratios. (C) Effect of BAY, FDI-6 and their combination on the colony formation of HUH7 cells. (D) Effect of BAY, FDI-6 and their combination on the apoptosis of HUH7 cells. (E) The CI values of the combinations of TST and FDI-6 at different concentration ratios. (F) Effect of TST, FDI-6 and their combination on the colony formation of HUH7 cells. (G) The CI values of the combinations of RCM-1 and FDI-6 at different concentration ratios. (H) Effect of RCM-1, FDI-6 and their combination on the colony formation of HUH7 cells. (I) Effect of BAY, FDI-6 and their sequential combination on tumor volume. (J) Effect of BAY, FDI-6 and their sequential combination on tumor weight. (K) The photos of HUH7 xenograft tumors in each group. (L) Acute toxicity analysis of 100 mg/kg FDI-6, 100 mg/kg BAY, and their combination in mice. (M) Effect of BAY, FDI-6 and their sequential combination on mouse weight. (N) Effect of BAY, FDI-6 and their sequential combination on the heart, liver, spleen, lung and kidney in mice. (O) Effect of BAY, FDI-6 and their sequential combination on the expression of Ki-67, FOXM1, and BUB1 in HUH7 xenograft tumors. Images were randomly selected from five replicates. Data from three independent experiments were statistically analyzed using one-way ANOVA: * P<0.05, ** P<0.01 vs. control group, # P < 0.05, ## P < 0.01 vs. BAY-FDI-6 group.

Techniques Used: Concentration Assay, Expressing, Control

FOXM1/BUB1 axis drives HCC cell DNA repair, G2/M transition, stemness, migration, and invasion. (A) Effect of BAY, FDI-6 and their combination on DNA damage in HUH7 cells. (B) Effect of BAY, FDI-6 and their combination on DNA repair-related genes in HUH7 cells analyzed by Q-PCR. (C) Effect of BAY, FDI-6 and their combination on cell cycle progression in HUH7 cells. (D) Effect of BAY, FDI-6 and their combination on cell cycle-related genes in HUH7 cells analyzed by Q-PCR. (E) Effect of BAY, FDI-6 and their combination on CD44 expression in HUH7 cells analyzed by IF. (F) Q-PCR analysis of the effects of BAY, FDI-6 and their combination on the expression of cell cycle-related genes in HUH7 cells. (G) Effect of BAY, FDI-6 and their combination on the formation of HUH7 spheres. (H) Effect of BAY, FDI-6 and their combination on the migration and invasion of HUH7 cells. (I) Q-PCR analysis of the effects of BAY, FDI-6 and their combination on the expression of EMT-related genes in HUH7 cells. (J) The molecular mechanism of the FOXM1/BUB1 axis in regulating HCC malignancy. Images were randomly selected from five replicates. Data from three independent experiments were statistically analyzed using one-way ANOVA: * P<0.05, ** P<0.01 vs. control group, # P < 0.05, ## P < 0.01 vs. BAY-FDI-6 group.

Figure Legend Snippet: FOXM1/BUB1 axis drives HCC cell DNA repair, G2/M transition, stemness, migration, and invasion. (A) Effect of BAY, FDI-6 and their combination on DNA damage in HUH7 cells. (B) Effect of BAY, FDI-6 and their combination on DNA repair-related genes in HUH7 cells analyzed by Q-PCR. (C) Effect of BAY, FDI-6 and their combination on cell cycle progression in HUH7 cells. (D) Effect of BAY, FDI-6 and their combination on cell cycle-related genes in HUH7 cells analyzed by Q-PCR. (E) Effect of BAY, FDI-6 and their combination on CD44 expression in HUH7 cells analyzed by IF. (F) Q-PCR analysis of the effects of BAY, FDI-6 and their combination on the expression of cell cycle-related genes in HUH7 cells. (G) Effect of BAY, FDI-6 and their combination on the formation of HUH7 spheres. (H) Effect of BAY, FDI-6 and their combination on the migration and invasion of HUH7 cells. (I) Q-PCR analysis of the effects of BAY, FDI-6 and their combination on the expression of EMT-related genes in HUH7 cells. (J) The molecular mechanism of the FOXM1/BUB1 axis in regulating HCC malignancy. Images were randomly selected from five replicates. Data from three independent experiments were statistically analyzed using one-way ANOVA: * P<0.05, ** P<0.01 vs. control group, # P < 0.05, ## P < 0.01 vs. BAY-FDI-6 group.

Techniques Used: Migration, Expressing, Control

Control:

Article Title: miR-149 Inhibits Non-Small-Cell Lung Cancer Cells EMT by Targeting FOXM1
Article Snippet: FOXM1-shRNA and the control shRNA were purchased from GeneChem (Shanghai, China). pcDNA3-FOXM1 was gifted by Dr. Sarkar at Wayne State University.

Article Title: miR-149 Inhibits Non-Small-Cell Lung Cancer Cells EMT by Targeting FOXM1.
Article Snippet: FOXM1-shRNA and the control shRNAwere purchased from GeneChem (Shanghai, China). pcDNA3-FOXM1 was gifted by Dr. Sarkar at Wayne State University.

shRNA:

Image Search Results

Journal: International Journal of Biological Sciences

Article Title: Mechanistic Insights into the FOXM1/BUB1 axis-Mediated Oncogenic Signaling in Hepatocellular Carcinoma

doi: 10.7150/ijbs.125454

Figure Lengend Snippet: FOXM1 promotes the proliferation of HCC cells in vitro . (A) The expression of FOXM1 in tumor tissues and adjacent tissues of patients with HCC analyzed by IHC. (B) The expression of Ki-67 in tumor tissues of patients with HCC analyzed by IHC. (C) The differential expression of FOXM1 between tumor tissues and adjacent tissues of patients with HCC analyzed by IHC. (D) Correlation analysis of FOXM1 and Ki-67 IHC scores in tumor tissues of patients with HCC. (E) Box plots for the differential expression of FOXM1 between LIHC tissues and adjacent tissues in TCGA. (F) ROC curves for the relationship between FOXM1 expression and the prognosis of HCC patients. (G) Correlation between FOXM1 expression levels and survival of HCC patients. (H) Distribution of FOXM1 mRNA expression in HCC cell lines. (I) Expression of FOXM1 in HUH7, HepG2, and SMMC-7721 cells analyzed by Western blot. (J) Effect of FOXM1 shRNA and FOXM1 cDNA on the viability of HUH7 cells. (K) Effects of FOXM1 on the formation of HUH7 colonies. (L) Effects of FOXM1 on the apoptosis of HUH7 cells. (M) Effects of FDI-6 on the proliferation of HUH7 cells. (N) Effects of FDI-6 on the formation of HUH7 colonies. (O) Effects of FOXM1 shRNA and FOXM1 cDNA on mouse weight. (P) Effects of FOXM1 shRNA and FOXM1 cDNA on tumor volume. (Q) The photos of tumor nodules in each group. (R) Effects of FOXM1 shRNA and FOXM1 cDNA on tumor weight. (S) Effects of FOXM1 shRNA and FOXM1 cDNA on tumor proliferation. (T) The expression of Ki-67 and FOXM1 in xenograft tumor analyzed by IHC. Images were randomly selected from five replicates. Data from three independent experiments were analyzed by one-way ANOVA: * P<0.05, ** P<0.01 vs. control group; # P<0.05, ## P<0.01 vs. FOXM1 KD group.

Article Snippet: Lentiviral recombination vectors encoding human FOXM1 complete DNA (FOXM1 cDNA) (pGV341-cFOXM1), human BUB1 complete DNA (BUB1 cDNA) (pGV341-cBUB1), empty vector control (pGV341-cNC), lentiviral shRNA vectors targeting FOXM1 (pGV112-shFOXM1), BUB1 (pGV112-shBUB1), and scrambled control (pGV112-shNC) were constructed and purchased from Genechem Co. Ltd. (Shanghai, China).

Techniques: In Vitro, Expressing, Quantitative Proteomics, Western Blot, shRNA, Control

Journal: International Journal of Biological Sciences

Article Title: Mechanistic Insights into the FOXM1/BUB1 axis-Mediated Oncogenic Signaling in Hepatocellular Carcinoma

doi: 10.7150/ijbs.125454

Figure Lengend Snippet: FOXM1-regulated genes involve in DNA repair, cell cycle, stemness and EMT. (A) GO function and KEGG pathway enrichment analysis of DEGs between FOXM1 KD and NC KD in HUH7 cells. (B) . GO function and KEGG pathway enrichment analysis of top 50 targets in the PPI network of FOXM1. (C) . Volcano plot for the DEGs involved in DNA repair and cell cycle analyzed by RNA-seq. (D) Volcano plot for the DEGs involved in cell stemness, invasion and migration. (E) . The relationship between FOXM1 expression and HCC cell stemness in TCGA. (F) . Differential expression of genes between LIHC tissues and adjacent tissues in TCGA. (G) . ROC curves for the relationship between gene expression and HCC prognosis. Data were statistically analyzed using one-way ANOVA: * P<0.05 compared with adjacent tissues.

Techniques: RNA Sequencing, Migration, Expressing, Quantitative Proteomics, Gene Expression

Journal: International Journal of Biological Sciences

Article Title: Mechanistic Insights into the FOXM1/BUB1 axis-Mediated Oncogenic Signaling in Hepatocellular Carcinoma

doi: 10.7150/ijbs.125454

Figure Lengend Snippet: FOXM1 promotes DNA repair and G2/M progression on HCC cells. (A) Effects of FOXM1 on DNA damage in HUH7 and HepG2 cells analyzed by comet assay. (B) Effects of FOXM1 on γH2AX expression analyzed by IF. (C) Q-PCR analysis of the effects of FOXM1 on the expression of DNA repair-related genes. (D) Analysis of the correlation between FOXM1 and DNA repair-related gene expressions in LIHC. (E) Effects of FOXM1 on cell cycle progression in HUH7 cells. (F) percentage of cell cycle at G0/G1, S, and G2/M phases. (G) Q-PCR analysis of the effects of FOXM1 on the expression of genes regulating G2/M transition. (H) Analysis of the correlation between FOXM1 and cell cycle related gene expressions in HCC. (I) Effects of FDI-6 on DNA damage in HUH7 cells. (J) Effects of FDI-6 on the expression of DNA-repair related genes in HUH7 cells. (K) Effects of FDI-6 on cell cycle progression in HUH7 cells. (L) Effects of FDI-6 on cell cycle-related genes in HUH7 cells. Images were randomly selected from five replicates. Data from three independent experiments were statistically analyzed using one-way ANOVA: * P < 0.05, ** P < 0.01 vs. NC KD group; # P < 0.05, ## P < 0.01 vs. FOXM1 KD group.

Techniques: Single Cell Gel Electrophoresis, Expressing

Journal: International Journal of Biological Sciences

Article Title: Mechanistic Insights into the FOXM1/BUB1 axis-Mediated Oncogenic Signaling in Hepatocellular Carcinoma

doi: 10.7150/ijbs.125454

Figure Lengend Snippet: FOXM1 drives cell stemness, invasion, and migration in HCC cells. (A) Effects of FOXM1 on CD44 expression analyzed by IF in HUH7 cells. (B) Effects of FOXM1 on CD44 expression in HepG2 cells. (C) Effects of FOXM1 on the expression of stemness-related genes in HUH7 cells. (D) Effects of FDI-6 on CD44 expression in HUH7 cells. (E) Effects of FDI-6 on the expression of stemness-related genes in HUH7 cells. (F) Effects of FOXM1 shRNA and FDI-6 on the formation of HUH7 three-dimensional spheres. (G) Analysis of the correlation between FOXM1 and cell cycle-related gene expressions in HCC. (H) Effects of FOXM1 on the invasion and migration of HUH7 and HepG2 cells. (I) Q-PCR analysis of the effects of FOXM1 on EMT-related gene expression in HUH7 cells. (J) Effects of FDI-6 on the invasion and migration of HUH7 cells. (K) Q-PCR analysis of the effects of FDI-6 on EMT-related gene expression in HUH7 cells. (L) Analysis of the correlation between FOXM1 and EMT-related gene expressions in HCC. Images were randomly selected from five replicates. Data from three independent experiments were statistically analyzed using one-way ANOVA: * P < 0.05, ** P < 0.01 vs. NC KD group; # P < 0.05, ## P < 0.01 vs. FOXM1 KD group.

Techniques: Migration, Expressing, shRNA, Gene Expression

Journal: International Journal of Biological Sciences

Article Title: Mechanistic Insights into the FOXM1/BUB1 axis-Mediated Oncogenic Signaling in Hepatocellular Carcinoma

doi: 10.7150/ijbs.125454

Figure Lengend Snippet: FOXM1 promotes BUB1 expression at transcriptional level. (A) Venn diagram of overlapping genes identified by RNA-seq and FOXM1 PPI network. (B) Analysis the association between key gene expression and HCC patient survival. (C) The expression of BUB1 between HCC tissues and adjacent tissues from clinical patients analyzed by IHC. (D) IHC scores of BUB1 between HCC tissues and adjacent tissues from clinical patients. (E) Correlation analysis of BUB1 and Ki-67 IHC scores in tumor tissues of patients with HCC. (F) Correlation analysis of BUB1 and FOXM1 IHC scores in tumor tissues of patients with HCC. (G) Co-IP analysis of the interaction between FOXM1 and BUB1 in HUH7 cells. (H) Effects of FOXM1 shRNA and BUB1 shRNA on the expression of FOXM1 and BUB1 analyzed by Q-PCR. (I) The expression of FOXM1 and BUB1 analyzed by Western blot. (J) ChIP-qPCR analysis of the binding of FOXM1 and BUB1 promoter in HUH7 cells. (K) The sequences at the -293 bp of the BUB1-WT promoter and the BUB1-Mut promoter. (L) The binding of FOXM1 and BUB1 promoter at the -293 bp sequence GTAAACC analyzed by dual luciferase reporter assay. (M) Q-PCR analysis of BUB1 expression in different HCC cells. (N) Western blot analysis of BUB1 expression in different HCC cells. (O) KEGG pathway enrichment analysis of top 50 targets in the PPI network of BUB1. (P) Venn diagram of overlapping genes identified by RNA-seq, FOXM1 PPI network, and BUB1 PPI network. (Q) correlation analysis of BUB1 expression and LIHC cell stemness in TCGA. (R) Effect of BUB1 on the inhibitory role of FOXM1 shRNA in the proliferation of HUH7 cells. (S) Effects of BUB1 on the inhibition of FOXM1 shRNA in HUH7 colony formation. (T) Effects of BUB1 on the promotion of FOXM1 shRNA in HUH7 cell apoptosis. (U) Effects of BUB1 shRNA on HUH7 xenograft tumor volumes. (V) Photos of HUH7 xenograft tumors in each group. (W) Effects of BUB1 shRNA on HUH7 xenograft tumor weight. (X) Effects of BUB1 shRNA on Ki-67 and BUB1 expression in each group. Images were randomly selected from five replicates. Data from three independent experiments were analyzed by one-way ANOVA: * P<0.05, ** P<0.01 vs. NC KD group, # P < 0.05, ## P < 0.01 vs. FOXM1 KD group.

Techniques: Expressing, RNA Sequencing, Gene Expression, Co-Immunoprecipitation Assay, shRNA, Western Blot, ChIP-qPCR, Binding Assay, Sequencing, Luciferase, Reporter Assay, Inhibition

Journal: International Journal of Biological Sciences

Article Title: Mechanistic Insights into the FOXM1/BUB1 axis-Mediated Oncogenic Signaling in Hepatocellular Carcinoma

doi: 10.7150/ijbs.125454

Figure Lengend Snippet: Knockdown of BUB1 enhances HCC cell sensitivity to FOXM1 inhibitor FDI-6. (A) Effect of BUB1 shRNA on FDI-6-mediated inhibition of colony formation in HUH7 cells. (B) Effect of BUB1 shRNA on FDI-6-induced apoptosis. (C) Effect of BUB1 shRNA on FDI-6-induced DNA damage. (D) Effect of BUB1 shRNA on the G2/M phase arrest caused by FDI-6. (E) Western blot analysis of BUB1 shRNA and FDI-6 effects on DNA repair-related gene expression. (F) Western blot analysis of BUB1 shRNA and FDI-6 effects on cell cycle-related gene expression. (G) IF analysis of BUB1 shRNA and FDI-6 effects on CD44 expression. (H) Western blot analysis of BUB1 shRNA and FDI-6 effects on stemness-related gene expression. (I) Effects of BUB1 shRNA on FDI-6 mediated suppression of HCC cell invasion and migration. (J) Western blot analysis of BUB1 shRNA and FDI-6 effects on EMT-related gene expression. (K) Effects of BUB1 shRNA and FDI-6 on mouse weight. (L) Effects of BUB1 shRNA and FDI-6 on tumor volume. (M) The photos of HUH7 xenograft tumors in each group. (N) Effects of BUB1 shRNA and FDI-6 on tumor weight. (O) The inhibition ratios of BUB1 shRNA and FDI-6 on tumor volume and weight. (P) IHC analysis of the effects of BUB1 shRNA and FDI-6 on Ki-67 and FOXM1 expression Images were randomly selected from five replicates. Data from three independent experiments were statistically analyzed using one-way ANOVA: * P<0.05, ** P<0.01 vs. NC KD group, # P < 0.05, ## P < 0.01 vs. BUB1 KD+FDI-6 group.

Techniques: Knockdown, shRNA, Inhibition, Western Blot, Gene Expression, Expressing, Migration

Journal: International Journal of Biological Sciences

Article Title: Mechanistic Insights into the FOXM1/BUB1 axis-Mediated Oncogenic Signaling in Hepatocellular Carcinoma

doi: 10.7150/ijbs.125454

Figure Lengend Snippet: FOXM1 inhibitors and BAY synergistically inhibit proliferation of HCC cells and tumors. (A) Effect of BAY and FDI-6 on the proliferation of HUH7 cells. (B) The CI values of the combinations of BAY and FDI-6 at different concentration ratios. (C) Effect of BAY, FDI-6 and their combination on the colony formation of HUH7 cells. (D) Effect of BAY, FDI-6 and their combination on the apoptosis of HUH7 cells. (E) The CI values of the combinations of TST and FDI-6 at different concentration ratios. (F) Effect of TST, FDI-6 and their combination on the colony formation of HUH7 cells. (G) The CI values of the combinations of RCM-1 and FDI-6 at different concentration ratios. (H) Effect of RCM-1, FDI-6 and their combination on the colony formation of HUH7 cells. (I) Effect of BAY, FDI-6 and their sequential combination on tumor volume. (J) Effect of BAY, FDI-6 and their sequential combination on tumor weight. (K) The photos of HUH7 xenograft tumors in each group. (L) Acute toxicity analysis of 100 mg/kg FDI-6, 100 mg/kg BAY, and their combination in mice. (M) Effect of BAY, FDI-6 and their sequential combination on mouse weight. (N) Effect of BAY, FDI-6 and their sequential combination on the heart, liver, spleen, lung and kidney in mice. (O) Effect of BAY, FDI-6 and their sequential combination on the expression of Ki-67, FOXM1, and BUB1 in HUH7 xenograft tumors. Images were randomly selected from five replicates. Data from three independent experiments were statistically analyzed using one-way ANOVA: * P<0.05, ** P<0.01 vs. control group, # P < 0.05, ## P < 0.01 vs. BAY-FDI-6 group.

Techniques: Concentration Assay, Expressing, Control

Journal: International Journal of Biological Sciences

Article Title: Mechanistic Insights into the FOXM1/BUB1 axis-Mediated Oncogenic Signaling in Hepatocellular Carcinoma

doi: 10.7150/ijbs.125454

Figure Lengend Snippet: FOXM1/BUB1 axis drives HCC cell DNA repair, G2/M transition, stemness, migration, and invasion. (A) Effect of BAY, FDI-6 and their combination on DNA damage in HUH7 cells. (B) Effect of BAY, FDI-6 and their combination on DNA repair-related genes in HUH7 cells analyzed by Q-PCR. (C) Effect of BAY, FDI-6 and their combination on cell cycle progression in HUH7 cells. (D) Effect of BAY, FDI-6 and their combination on cell cycle-related genes in HUH7 cells analyzed by Q-PCR. (E) Effect of BAY, FDI-6 and their combination on CD44 expression in HUH7 cells analyzed by IF. (F) Q-PCR analysis of the effects of BAY, FDI-6 and their combination on the expression of cell cycle-related genes in HUH7 cells. (G) Effect of BAY, FDI-6 and their combination on the formation of HUH7 spheres. (H) Effect of BAY, FDI-6 and their combination on the migration and invasion of HUH7 cells. (I) Q-PCR analysis of the effects of BAY, FDI-6 and their combination on the expression of EMT-related genes in HUH7 cells. (J) The molecular mechanism of the FOXM1/BUB1 axis in regulating HCC malignancy. Images were randomly selected from five replicates. Data from three independent experiments were statistically analyzed using one-way ANOVA: * P<0.05, ** P<0.01 vs. control group, # P < 0.05, ## P < 0.01 vs. BAY-FDI-6 group.

Techniques: Migration, Expressing, Control

CDK7 inhibition elevates luminal‐related transcriptional activities and renders TNBC more sensitive to abemaciclib. A) The box plots depicting the dependency score of CDK7 and several other CDKs in TNBC cells ( n = 25) using CRISPR screening datasets from the Broad Institute DepMap portal. p values were calculated using one‐way ANOVA, **** p < 0.0001. B) Violin plots of CDK7 dependency scores in TNBC and non‐TNBC breast cancer cells ( n = 24 for TNBC, n = 17 for non‐TNBC) using CRISPR and RNAi screening datasets from the Broad Institute DepMap portal. p values were calculated using unpaired t ‐test, * p < 0.05,** p < 0.01. C) Violin plots of CDK7 mRNA expression levels in the TCGA‐TNBC dataset ( n = 140) and normal breast tissues from the GTEx dataset ( n = 459). P values were calculated using an unpaired t ‐test, **** p < 0.0001. D) Kaplan–Meier plots of CDK7 expression in TNBC patients using the TCGA cohort and KMplot cohort. Data were analyzed using the log‐rank test. E,F) Gene set enrichment analysis (GSEA) of RNA‐Seq data for CDK7‐high and CDK7‐low patients in the TCGA‐TNBC (E) and FUSCC‐TNBC (F) cohorts. NES, normalized enrichment score, NOM, nominal, FDR, false discovery rate. G) Heatmap summarizing the RNA‐Seq data of selected luminal/epithelial marker genes and basal/invasive marker genes in normal control (NC) and CDK7‐knockdown (ShCDK7) Hs578T cells ( n = 3). H) Immunoblot validation of CDK7 knockdown in MDA‐MB‐468 and Hs578T cells. I) Differences in drug sensitivities between MDA‐MB‐468‐NC and MDA‐MB‐468‐ShCDK7 cells, as well as between Hs578T‐NC and Hs578T‐ShCDK7 cells. Data are mean ± SD of 5 replicates. J,K) Dose‐response curves of tamoxifen (J) and abemaciclib (K) between NC and ShCDK7 MDA‐MB‐468 and Hs578T cells. Data are mean ± SD of 3–5 experimental replicates. p values were analyzed using a two‐way ANOVA test with Bonferroni correction.

Journal: Advanced Science

Article Title: Dual Inhibition of CDK4/6 and CDK7 Suppresses Triple‐Negative Breast Cancer Progression via Epigenetic Modulation of SREBP1‐Regulated Cholesterol Metabolism

doi: 10.1002/advs.202413103

Figure Lengend Snippet: CDK7 inhibition elevates luminal‐related transcriptional activities and renders TNBC more sensitive to abemaciclib. A) The box plots depicting the dependency score of CDK7 and several other CDKs in TNBC cells ( n = 25) using CRISPR screening datasets from the Broad Institute DepMap portal. p values were calculated using one‐way ANOVA, **** p < 0.0001. B) Violin plots of CDK7 dependency scores in TNBC and non‐TNBC breast cancer cells ( n = 24 for TNBC, n = 17 for non‐TNBC) using CRISPR and RNAi screening datasets from the Broad Institute DepMap portal. p values were calculated using unpaired t ‐test, * p < 0.05,** p < 0.01. C) Violin plots of CDK7 mRNA expression levels in the TCGA‐TNBC dataset ( n = 140) and normal breast tissues from the GTEx dataset ( n = 459). P values were calculated using an unpaired t ‐test, **** p < 0.0001. D) Kaplan–Meier plots of CDK7 expression in TNBC patients using the TCGA cohort and KMplot cohort. Data were analyzed using the log‐rank test. E,F) Gene set enrichment analysis (GSEA) of RNA‐Seq data for CDK7‐high and CDK7‐low patients in the TCGA‐TNBC (E) and FUSCC‐TNBC (F) cohorts. NES, normalized enrichment score, NOM, nominal, FDR, false discovery rate. G) Heatmap summarizing the RNA‐Seq data of selected luminal/epithelial marker genes and basal/invasive marker genes in normal control (NC) and CDK7‐knockdown (ShCDK7) Hs578T cells ( n = 3). H) Immunoblot validation of CDK7 knockdown in MDA‐MB‐468 and Hs578T cells. I) Differences in drug sensitivities between MDA‐MB‐468‐NC and MDA‐MB‐468‐ShCDK7 cells, as well as between Hs578T‐NC and Hs578T‐ShCDK7 cells. Data are mean ± SD of 5 replicates. J,K) Dose‐response curves of tamoxifen (J) and abemaciclib (K) between NC and ShCDK7 MDA‐MB‐468 and Hs578T cells. Data are mean ± SD of 3–5 experimental replicates. p values were analyzed using a two‐way ANOVA test with Bonferroni correction.

Article Snippet: Human shRNA lentiviruses targeting CDK7 (GIEL0325417) and FOXM1 (GIEE0370796) were obtained from GeneChem.

Techniques: Inhibition, CRISPR, Expressing, RNA Sequencing, Marker, Control, Knockdown, Western Blot, Biomarker Discovery

$The synergistic lethality of co‐inhibiting CDK4/6 and CDK7 in TNBC. A) Colony formation images of TNBC cells following a 48 h exposure to the combination of abemaciclib with YKL‐5‐124. Representative images from 3 biological replicates are provided. B) Colony formation images of TNBC cells following a 48 h exposure to the combination of palbociclib with YKL‐5‐124. Representative images from 3 biological replicates are provided. C) Heatmap of survival fractions in TNBC cells after 48 h exposure to gradient concentrations of abemaciclib, YKL‐5‐124, and the combined treatment (abemaciclib at gradient concentrations with YKL‐5‐124 at fixed concentrations). Data are shown as mean ( n = 3 biological replicates). D) Combination index values for TNBC cells treated with abemaciclib plus YKL‐5‐124, calculated by CompuSyn software. Data are represented as mean ± SD. E) Tumor weights of MDA‐MB‐468 and patient‐derived xenografts (PDX) after 21 days of treatment with control, abemaciclib (50 mg kg −1 ), YKL‐5‐124 (2 mg kg −1 ), or the combination ( n = 5). Tumor weights of MDA‐MB‐231 xenografts after 14 days of treatment with control, abemaciclib (50 mg kg −1 ), YKL‐5‐124 (5 mg kg −1 ), or the combination ( n = 6). Data are shown as mean ± SD. p values were calculated using one‐way ANOVA, * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001. F) Immunohistochemistry (IHC) staining of ki‐67 in tumor sections of MDA‐MB‐468, MDA‐MB‐231, and patient‐derived xenografts. Scale bar, 100 µm. G) Quantifications of ki‐67 staining in tumor sections of MDA‐MB‐468 ( n = 10), MDA‐MB‐231 ( n = 12), and patient‐derived xenografts ( n = 10). Two representative images per tumor were used to quantify the ki‐67 positivity. Data are represented as mean ± SD. p values were calculated using one‐way ANOVA, * p < 0.05, **** p < 0.0001. H) Percentage of total apoptotic cells after 96 h of treatment with abemaciclib, YKL‐5‐124, and the combination ( n = 3). Data are presented as mean ± SD. p values were calculated using one‐way ANOVA, **** p < 0.0001. I) IHC staining of cleaved caspase‐3 in tumor sections of MDA‐MB‐468, MDA‐MB‐231, and patient‐derived xenografts. Scale bar, 100 µm. J) H‐scores of cleaved caspase‐3 staining in tumor sections of MDA‐MB‐468 ( n = 10), MDA‐MB‐231 ( n = 12), and patient‐derived xenografts ( n = 10). Two representative images per tumor were used to quantify cleaved caspase‐3 staining. Data are represented as mean ± SD. p values were calculated using one‐way ANOVA, **** p < 0.0001.$

Journal: Advanced Science

Article Title: Dual Inhibition of CDK4/6 and CDK7 Suppresses Triple‐Negative Breast Cancer Progression via Epigenetic Modulation of SREBP1‐Regulated Cholesterol Metabolism

doi: 10.1002/advs.202413103

Figure Lengend Snippet: The synergistic lethality of co‐inhibiting CDK4/6 and CDK7 in TNBC. A) Colony formation images of TNBC cells following a 48 h exposure to the combination of abemaciclib with YKL‐5‐124. Representative images from 3 biological replicates are provided. B) Colony formation images of TNBC cells following a 48 h exposure to the combination of palbociclib with YKL‐5‐124. Representative images from 3 biological replicates are provided. C) Heatmap of survival fractions in TNBC cells after 48 h exposure to gradient concentrations of abemaciclib, YKL‐5‐124, and the combined treatment (abemaciclib at gradient concentrations with YKL‐5‐124 at fixed concentrations). Data are shown as mean ( n = 3 biological replicates). D) Combination index values for TNBC cells treated with abemaciclib plus YKL‐5‐124, calculated by CompuSyn software. Data are represented as mean ± SD. E) Tumor weights of MDA‐MB‐468 and patient‐derived xenografts (PDX) after 21 days of treatment with control, abemaciclib (50 mg kg −1 ), YKL‐5‐124 (2 mg kg −1 ), or the combination ( n = 5). Tumor weights of MDA‐MB‐231 xenografts after 14 days of treatment with control, abemaciclib (50 mg kg −1 ), YKL‐5‐124 (5 mg kg −1 ), or the combination ( n = 6). Data are shown as mean ± SD. p values were calculated using one‐way ANOVA, * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001. F) Immunohistochemistry (IHC) staining of ki‐67 in tumor sections of MDA‐MB‐468, MDA‐MB‐231, and patient‐derived xenografts. Scale bar, 100 µm. G) Quantifications of ki‐67 staining in tumor sections of MDA‐MB‐468 ( n = 10), MDA‐MB‐231 ( n = 12), and patient‐derived xenografts ( n = 10). Two representative images per tumor were used to quantify the ki‐67 positivity. Data are represented as mean ± SD. p values were calculated using one‐way ANOVA, * p < 0.05, **** p < 0.0001. H) Percentage of total apoptotic cells after 96 h of treatment with abemaciclib, YKL‐5‐124, and the combination ( n = 3). Data are presented as mean ± SD. p values were calculated using one‐way ANOVA, **** p < 0.0001. I) IHC staining of cleaved caspase‐3 in tumor sections of MDA‐MB‐468, MDA‐MB‐231, and patient‐derived xenografts. Scale bar, 100 µm. J) H‐scores of cleaved caspase‐3 staining in tumor sections of MDA‐MB‐468 ( n = 10), MDA‐MB‐231 ( n = 12), and patient‐derived xenografts ( n = 10). Two representative images per tumor were used to quantify cleaved caspase‐3 staining. Data are represented as mean ± SD. p values were calculated using one‐way ANOVA, **** p < 0.0001.

Article Snippet: Human shRNA lentiviruses targeting CDK7 (GIEL0325417) and FOXM1 (GIEE0370796) were obtained from GeneChem.

Techniques: Software, Derivative Assay, Control, Immunohistochemistry, Staining

Concurrent inhibition of CDK4/6 and CDK7 suppresses SREBF1‐regulated cholesterol synthesis. A) Venn diagram illustrating the overlapping GSEA hallmark pathways that are enriched in the DMSO groups of MDA‐MB‐468 and Hs578T cells compared to the drug combination (Combo) groups. GSEA was performed using the RNA‐Seq data after 48 h of treatment with DMSO, abemaciclib, YKL‐5‐124, or the combination ( n = 2 biological replicates). B) The overlapping hallmark pathways ranked by mean NES of MDA‐MB‐468 and Hs578T cells. C) GSEA enrichment plots of cholesterol‐related pathways in the DMSO groups of MDA‐MB‐468 cells. D) Schematic diagram of the cholesterol biosynthesis pathway, including intermediate metabolites (black) and key enzymes (blue). E) Heatmap summarizing the RT‐qPCR results of cholesterol synthesis‐related genes following single‐agent or combined treatment across two TNBC cell lines ( n = 3). F) Immunoblot analysis of cholesterol synthesis‐related proteins following single‐agent or combined treatment across two TNBC cell lines. G) IHC staining of SREBP1 in tumor sections of MDA‐MB‐468 and patient‐derived xenografts. Scale bar, 100 µm. H) H‐scores of SREBP1 staining in tumor sections of MDA‐MB‐468 ( n = 10) and patient‐derived xenografts ( n = 10). Two representative images per tumor were used to quantify SREBP1 staining. Data are represented as mean ± SD. p values were calculated using one‐way ANOVA, **** p < 0.0001. I) Quantification of cholesterol‐related metabolites in MDA‐MB‐468 xenografts ( n = 5) and Hs578T cells ( n = 5). Data are presented as mean ± SD. p values were calculated using one‐way ANOVA, * p < 0.05, ** p <0.01, **** p < 0.0001. J) Cholesterol rescued colony formation of combination groups in MDA‐MB‐468 and Hs578T cells. Rescue groups were additionally supplemented with 0.2 µg mL −1 cholesterol for 14 days. Representative images from 3 biological replicates are provided. p values were calculated using one‐way ANOVA, * p < 0.05, **** p < 0.0001. K) Schematic illustration of the in vivo cholesterol rescue experiment. Mice bearing MDA‐MB‐468 xenograft tumors were randomized to receive a control or a 1.25% cholesterol‐enriched diet, with or without the combination therapy (50 mg kg −1 abemaciclib plus 2 mg kg −1 YKL‐5‐124). Tumor weights at the study endpoint for the four treatment arms were collected: control with a chow diet, control with a 1.25% cholesterol diet, combined treatments with a chow diet, and combined treatments with a 1.25% cholesterol diet ( n = 5). Data are shown as mean ± SD. p values for tumor weights were determined using a two‐tailed Student's t ‐test, *** p < 0.001. L) Effects of SREBF1 overexpression (OE) upon drug synergy. Left, heatmaps of viability in SREBF1‐NC and SREBF1‐OE cells after a 48 h exposure to the indicated concentrations of abemaciclib, YKL‐5‐124, and the combined treatment (gradient concentrations of abemaciclib in combination with 1 µM YKL‐5‐124). Data are presented as the mean values from three biological replicates. Right, combination index values for SREBF1‐NC and SREBF1‐OE cells after the combined treatments. Data are shown as mean ± SD. p values were determined using an unpaired t ‐test, **** p < 0.0001.

Journal: Advanced Science

Article Title: Dual Inhibition of CDK4/6 and CDK7 Suppresses Triple‐Negative Breast Cancer Progression via Epigenetic Modulation of SREBP1‐Regulated Cholesterol Metabolism

doi: 10.1002/advs.202413103

Figure Lengend Snippet: Concurrent inhibition of CDK4/6 and CDK7 suppresses SREBF1‐regulated cholesterol synthesis. A) Venn diagram illustrating the overlapping GSEA hallmark pathways that are enriched in the DMSO groups of MDA‐MB‐468 and Hs578T cells compared to the drug combination (Combo) groups. GSEA was performed using the RNA‐Seq data after 48 h of treatment with DMSO, abemaciclib, YKL‐5‐124, or the combination ( n = 2 biological replicates). B) The overlapping hallmark pathways ranked by mean NES of MDA‐MB‐468 and Hs578T cells. C) GSEA enrichment plots of cholesterol‐related pathways in the DMSO groups of MDA‐MB‐468 cells. D) Schematic diagram of the cholesterol biosynthesis pathway, including intermediate metabolites (black) and key enzymes (blue). E) Heatmap summarizing the RT‐qPCR results of cholesterol synthesis‐related genes following single‐agent or combined treatment across two TNBC cell lines ( n = 3). F) Immunoblot analysis of cholesterol synthesis‐related proteins following single‐agent or combined treatment across two TNBC cell lines. G) IHC staining of SREBP1 in tumor sections of MDA‐MB‐468 and patient‐derived xenografts. Scale bar, 100 µm. H) H‐scores of SREBP1 staining in tumor sections of MDA‐MB‐468 ( n = 10) and patient‐derived xenografts ( n = 10). Two representative images per tumor were used to quantify SREBP1 staining. Data are represented as mean ± SD. p values were calculated using one‐way ANOVA, **** p < 0.0001. I) Quantification of cholesterol‐related metabolites in MDA‐MB‐468 xenografts ( n = 5) and Hs578T cells ( n = 5). Data are presented as mean ± SD. p values were calculated using one‐way ANOVA, * p < 0.05, ** p <0.01, **** p < 0.0001. J) Cholesterol rescued colony formation of combination groups in MDA‐MB‐468 and Hs578T cells. Rescue groups were additionally supplemented with 0.2 µg mL −1 cholesterol for 14 days. Representative images from 3 biological replicates are provided. p values were calculated using one‐way ANOVA, * p < 0.05, **** p < 0.0001. K) Schematic illustration of the in vivo cholesterol rescue experiment. Mice bearing MDA‐MB‐468 xenograft tumors were randomized to receive a control or a 1.25% cholesterol‐enriched diet, with or without the combination therapy (50 mg kg −1 abemaciclib plus 2 mg kg −1 YKL‐5‐124). Tumor weights at the study endpoint for the four treatment arms were collected: control with a chow diet, control with a 1.25% cholesterol diet, combined treatments with a chow diet, and combined treatments with a 1.25% cholesterol diet ( n = 5). Data are shown as mean ± SD. p values for tumor weights were determined using a two‐tailed Student's t ‐test, *** p < 0.001. L) Effects of SREBF1 overexpression (OE) upon drug synergy. Left, heatmaps of viability in SREBF1‐NC and SREBF1‐OE cells after a 48 h exposure to the indicated concentrations of abemaciclib, YKL‐5‐124, and the combined treatment (gradient concentrations of abemaciclib in combination with 1 µM YKL‐5‐124). Data are presented as the mean values from three biological replicates. Right, combination index values for SREBF1‐NC and SREBF1‐OE cells after the combined treatments. Data are shown as mean ± SD. p values were determined using an unpaired t ‐test, **** p < 0.0001.

Article Snippet: Human shRNA lentiviruses targeting CDK7 (GIEL0325417) and FOXM1 (GIEE0370796) were obtained from GeneChem.

Techniques: Inhibition, RNA Sequencing, Quantitative RT-PCR, Western Blot, Immunohistochemistry, Derivative Assay, Staining, In Vivo, Control, Two Tailed Test, Over Expression

Clinical relevance of the SREBP1‐p300‐cholesterol synthesis pathway in TNBC. A,B) Correlation analysis between the mRNA expression levels of SREBF1, EP300, and cholesterol biosynthesis genes (PMVK, SQLE, and LSS) in the FUSCC‐TNBC ( n = 360) (A) and TCGA‐TNBC ( n = 140) (B) cohorts. Correlation coefficients were calculated using the Spearman test. p values were obtained using spearman correlation test. C) Kaplan–Meier plots of FOXM1, SREBF1, and EP300 expression in the FUSCC‐TNBC cohort. Data were analyzed using the log‐rank test. D) Kaplan–Meier plots of cholesterol homeostasis gene set variation analysis (GSVA) scores in the FUSCC‐TNBC cohort. Data were analyzed using the log‐rank test. E) Schematic diagram illustrating the proposed mechanism of co‐inhibiting CDK4/6 and CDK7 suppresses SREBP1‐regulated cholesterol biosynthesis. Reduced CDK7 expression disrupts luminal and basal transcriptional activities within TNBC, enabling tumors to overcome intrinsic resistance to CDK4/6 inhibitors. The synergistic intervention initially diminishes the activation of FOXM1, which directly binds with the promoter of SREBF1, exerting regulatory control over SREBF1 transcriptional activity. Consequently, SREBF1 mRNA and protein levels are decreased, attenuating SREBP1/p300 co‐recruitment to cholesterol synthesis gene promoters. This cascade transcriptionally represses rate‐limiting cholesterogenic enzymes, lowers cholesterol synthesis, and maintains antitumor effects.

Journal: Advanced Science

Article Title: Dual Inhibition of CDK4/6 and CDK7 Suppresses Triple‐Negative Breast Cancer Progression via Epigenetic Modulation of SREBP1‐Regulated Cholesterol Metabolism

doi: 10.1002/advs.202413103

Figure Lengend Snippet: Clinical relevance of the SREBP1‐p300‐cholesterol synthesis pathway in TNBC. A,B) Correlation analysis between the mRNA expression levels of SREBF1, EP300, and cholesterol biosynthesis genes (PMVK, SQLE, and LSS) in the FUSCC‐TNBC ( n = 360) (A) and TCGA‐TNBC ( n = 140) (B) cohorts. Correlation coefficients were calculated using the Spearman test. p values were obtained using spearman correlation test. C) Kaplan–Meier plots of FOXM1, SREBF1, and EP300 expression in the FUSCC‐TNBC cohort. Data were analyzed using the log‐rank test. D) Kaplan–Meier plots of cholesterol homeostasis gene set variation analysis (GSVA) scores in the FUSCC‐TNBC cohort. Data were analyzed using the log‐rank test. E) Schematic diagram illustrating the proposed mechanism of co‐inhibiting CDK4/6 and CDK7 suppresses SREBP1‐regulated cholesterol biosynthesis. Reduced CDK7 expression disrupts luminal and basal transcriptional activities within TNBC, enabling tumors to overcome intrinsic resistance to CDK4/6 inhibitors. The synergistic intervention initially diminishes the activation of FOXM1, which directly binds with the promoter of SREBF1, exerting regulatory control over SREBF1 transcriptional activity. Consequently, SREBF1 mRNA and protein levels are decreased, attenuating SREBP1/p300 co‐recruitment to cholesterol synthesis gene promoters. This cascade transcriptionally represses rate‐limiting cholesterogenic enzymes, lowers cholesterol synthesis, and maintains antitumor effects.

Article Snippet: Human shRNA lentiviruses targeting CDK7 (GIEL0325417) and FOXM1 (GIEE0370796) were obtained from GeneChem.

Techniques: Expressing, Activation Assay, Control, Activity Assay

Review

lentiviral shrna vectors targeting foxm1 pgv112 shfoxm1 (Genechem)

Structured Review

Images

1) Product Images from "Mechanistic Insights into the FOXM1/BUB1 axis-Mediated Oncogenic Signaling in Hepatocellular Carcinoma"

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