erbb2 overexpression lentivirus vectors  (Genechem)

Journal: Frontiers in Immunology

Article Title: The combination treatment of RC48 and STAT3 inhibitor acts as a promising therapeutic strategy for basal bladder cancer

doi: 10.3389/fimmu.2024.1432586

Figure Lengend Snippet: ERBB2 expression in different molecular types of MIBC. (A) Schematic diagram of consensus types of MIBC, including basal/squamous (Ba/Sq), luminal unstable (LumU), luminal nonspecified (LumNS), luminal papillary (LumP), neuroendocrine-like (NE-like), and stoma-rich. (B) ERBB2 mRNA expression in six MIBC consensus subtypes in the TCGA-MIBC dataset. (C) ERBB2 mRNA expression in luminal and basal cells in CCLE dataset. (D) The luminal and basal-type single-cell clusters were determined by their molecular markers (luminal: FOXA1 and GATA3; basal: CD44 and KRT5), and the distribution of ERBB2 mRNA expression in luminal and basal clusters is shown. (E) The luminal and basal bladder cancer tissues’ IHC were classified by their molecular marker (luminal: KRT20 and GATA3; basal: CD44 and KRT5/6), and ERBB2 protein expression was detected. (F) Quantification of ERBB2 immunohistochemistry protein levels in luminal and basal bladder cancer. (G) Western blot analysis showed the protein expression of ERBB2, KRT20, GATA3, CD44, and KRT5/6 in luminal (HT1376, RT4, and SW780) and basal (5637, J82, and T24) cells. (H) Quantitative chart of ERBB2, KRT20, GATA3, CD44, and KRT5/6 from the Western blot analysis. *, P-value < 0.05; **, P-value < 0.01; ***, P-value < 0.001; ****, P-value < 0.0001.

Article Snippet: The ERBB2 overexpression lentivirus vectors were purchased from Genechem (Shanghai, China).

Techniques: Expressing, Marker, Immunohistochemistry, Western Blot

Journal: Frontiers in Immunology

Article Title: The combination treatment of RC48 and STAT3 inhibitor acts as a promising therapeutic strategy for basal bladder cancer

doi: 10.3389/fimmu.2024.1432586

Figure Lengend Snippet: Luminal cells show higher sensitivity to RC48. (A) The Western blot shows ERBB2 protein expression levels in luminal cells (SW780, HT1376, and RT4) after exposure to RC48 at different concentrations and times. (B) The barplot shows the gDS in MIBC cells representing luminal and basal cells. (C) The percentage of surviving cells after exposing different concentrations of RC48 in three luminal (SW780, RT4, and HT1376) and three basal (J82, T24, and 5637) cells. (D) Colon formation assay reveals the cell growth after exposing different concentrations of RC48 in luminal and basal cells. The percentage of survival cells was quantified. (E) Cell viability of three luminal and three basal cells after exposing RC48 (25 μg/ml) at different times. *, P-value < 0.05; **, P-value < 0.01; ***, P-value < 0.001; ****, P-value < 0.0001; ns, not significant.

Article Snippet: The ERBB2 overexpression lentivirus vectors were purchased from Genechem (Shanghai, China).

Techniques: Western Blot, Expressing, Tube Formation Assay

Journal: Frontiers in Immunology

Article Title: The combination treatment of RC48 and STAT3 inhibitor acts as a promising therapeutic strategy for basal bladder cancer

doi: 10.3389/fimmu.2024.1432586

Figure Lengend Snippet: ERBB2 expression mediates the sensitivity of luminal and basal cells to RC48. (A) Western blot analysis validated the ERBB2 expression in ERBB2-silenced luminal cells (SW780 and HT1376) and ERBB2-overexpressing basal (T24 and 5637) cells. (B , C) The percentage of surviving cells after exposure to different concentrations of RC48 in ERBB2-silenced luminal cells (SW780 and HT1376)/or ERBB2-overexpressing basal cells (T24 and 5637). (D) Colon formation assay demonstrates cell growth after exposure to RC48 (5 μg/ml) in ERBB2-overexpressing basal cells. (E , F) Cell viability of the ERBB2-overexpressing and control basal cells after exposure to RC48 (25 μg/ml). (G) Tumors with different treatments were shown. (H) Tumor weight was measured after 4 weeks. Data are presented as the mean ± SD. (I) Tumor volume was measured one to two times weekly once the tumor became visible. Data are presented as the mean ± SD. (J) IHC revealed ERBB2 expression in tumor samples from mice receiving different treatments. *, P-value < 0.05; **, P-value < 0.01; ***, P-value < 0.001; ****, P-value < 0.0001.

Article Snippet: The ERBB2 overexpression lentivirus vectors were purchased from Genechem (Shanghai, China).

Techniques: Expressing, Western Blot, Tube Formation Assay, Control

Journal: Frontiers in Immunology

Article Title: The combination treatment of RC48 and STAT3 inhibitor acts as a promising therapeutic strategy for basal bladder cancer

doi: 10.3389/fimmu.2024.1432586

Figure Lengend Snippet: The STAT3 pathway is compensatively activated in basal cells. (A) The diagram shows the GESA result based on the DEGs between basal/squamous and the other three luminal consensus subtypes. (B) The Venn diagram shows the common altered pathways from the GSEA results based on the DEGs determined between basal and luminal types of four molecular classification systems. (C, D) The heatmap shows the expression of genes positively correlated with STAT3 pathways in basal and luminal bladder cancer tissues and cell lines. (E) STAT3 mRNA expression levels in consensus types of TCGA-MIBC. (F) STAT3 mRNA expression levels in luminal and basal cells. (G) STAT3 protein levels in luminal and basal cells were detected by Western blot analysis. (H) The diagram shows the GSEA results based on the DEGs between RC48-exposed SW780 and control. The top 10 activated pathways are shown. (I) The diagram shows the correlation between ERBB2 and STAT3 in mRNA levels, with the PCC used for analysis. (J, K) In ERBB2-silenced luminal cells and ERBB2-overexpressing basal cells, Western blot analysis was performed to measure the expression changes of ERBB2, STAT3, and pSTAT3 (Tyr705). (L) After exposure to RC48 (HT1376 and SW780), the proteins ERBB2, STAT3, and pSTAT3 (Tyr705) were detected by Western blot analysis. (M) Control and RC48-exposed SW780 and HT1376 cells, or ERBB2-overexpressing T24 and 5637 cells, were fixed and incubated with antibodies against pSTAT3 (Tyr705) for immunofluorescence analysis. *, P-value < 0.05; **, P-value < 0.01; ***, P-value < 0.001; ****, P-value < 0.0001.

Article Snippet: The ERBB2 overexpression lentivirus vectors were purchased from Genechem (Shanghai, China).

Techniques: Expressing, Western Blot, Control, Incubation, Immunofluorescence

Journal: Frontiers in Immunology

Article Title: The combination treatment of RC48 and STAT3 inhibitor acts as a promising therapeutic strategy for basal bladder cancer

doi: 10.3389/fimmu.2024.1432586

Figure Lengend Snippet: The combination of ART and RC48 blocks tumor growth in the xenograft model of nude mice. (A) Schematic diagram of the treatment paradigm. (B) Tumors with different treatments are shown. (C) Tumor weight was measured after 4 weeks. Data are presented as the mean ± SD. (D) Tumor volume was measured one to two times per week when the tumor became visible. Data are presented as the mean ± SD. (E) The Kaplan–Meier survival curve shows the prognostic differences among the control, RC48, artesunate, and RC48+artesunate groups. (F) The diagram shows changes in mouse body weight. (G) IHC shows the STAT3 expression in tumor samples from mice with different treatments. (H) Diagram of STAT3 pathways after treatment with RC48 in luminal and basal cells. (I) A combination of STAT3 and ERBB2 inhibition in vivo and mechanism diagram. *, P-value < 0.05; **, P-value < 0.01; ***, P-value < 0.001; ****, P-value < 0.0001; ns, not significant.

Article Snippet: The ERBB2 overexpression lentivirus vectors were purchased from Genechem (Shanghai, China).

Techniques: Control, Expressing, Inhibition, In Vivo