Journal: Cancer Medicine

Article Title: Nuclear receptor coactivator 6 ( NCoA6 ) promotes cell proliferation, migration, and invasion in pancreatic cancer

doi: 10.1002/cam4.6427

Figure Lengend Snippet: Upregulation of NCoA6 expression is correlated with PDAC progression. (A) NCoA6 expression is significantly increased in PDAC samples compared with normal tissues from TCGA and GTEx. (B) Representative images of IHC staining for NCoA6 in PDAC tumor tissues (T) and adjacent normal tissues (ANT). (C) IHC score of NCoA6 expression in PDAC and ANT ( n = 55, p < 0.001). (D) Representative images of IHC staining for NCoA6 in tissue microarrays. (E) Kaplan–Meier analysis of NCoA6 expression and overall survival ( p < 0.01) and disease‐free survival ( p < 0.05) of patients with PDAC from FUSCC. (F) The high NCoA6 expression group is associated with worse overall and disease‐free survival according to the Kaplan–Meier plotter database (hazard ratio [HR] = 1.44 and 3.15, respectively). (G) Time‐dependent ROC curve shows that NCoA6 expression in PDAC tissues from FUSCC is related to the 2‐year overall and disease‐free survival of patients (AUC = 0.822 and 0.707, respectively). (H) The ROC analysis from UCSC shows the moderate prognostic value of NCoA6 expression level in PDAC tissues (AUC = 0.811, p < 0.001).

Article Snippet: Incubation with primary antibodies for NCoA6 (1:100, Proteintech) and FBW7 (1:200, Bethyl) was at 4°C overnight, followed by polymeric HRP‐labeled anti‐rabbit IgG (Boster, China) at 37°C for 30 min. Tissue slides were scored to assess protein expression.

Techniques: Expressing, Immunohistochemistry

Journal: Cancer Medicine

Article Title: Nuclear receptor coactivator 6 ( NCoA6 ) promotes cell proliferation, migration, and invasion in pancreatic cancer

doi: 10.1002/cam4.6427

Figure Lengend Snippet: Relationship between clinicopathological features and NCoA6 expression in patients with pancreatic cancer.

Article Snippet: Incubation with primary antibodies for NCoA6 (1:100, Proteintech) and FBW7 (1:200, Bethyl) was at 4°C overnight, followed by polymeric HRP‐labeled anti‐rabbit IgG (Boster, China) at 37°C for 30 min. Tissue slides were scored to assess protein expression.

Techniques: Expressing

Journal: Cancer Medicine

Article Title: Nuclear receptor coactivator 6 ( NCoA6 ) promotes cell proliferation, migration, and invasion in pancreatic cancer

doi: 10.1002/cam4.6427

Figure Lengend Snippet: Univariate and multivariate Cox regression of overall survival and disease‐free survival of patients with PDAC.

Article Snippet: Incubation with primary antibodies for NCoA6 (1:100, Proteintech) and FBW7 (1:200, Bethyl) was at 4°C overnight, followed by polymeric HRP‐labeled anti‐rabbit IgG (Boster, China) at 37°C for 30 min. Tissue slides were scored to assess protein expression.

Techniques: Expressing

Journal: Cancer Medicine

Article Title: Nuclear receptor coactivator 6 ( NCoA6 ) promotes cell proliferation, migration, and invasion in pancreatic cancer

doi: 10.1002/cam4.6427

Figure Lengend Snippet: GO/KEGG analysis and GSEA of our RNA‐sequencing data. (A) Western blotting analysis of NCoA6 in PDAC lines. The H6C7 cell line was included as a negative control for the detection of endogenous NCoA6 expression, and β‐Actin was used as a control. (B) RT‐qPCR and (C) western blotting analysis of NCoA6 expression in NCoA6 PANC‐1 and SW1900 cells. The volcano plot (D) and heatmap (E) of gene alterations for NCoA6‐NC/sh PANC‐1 cells. (F) GSEA shows that SENESE_HDAC1_TARGETS_UP, the gene set from Human MSigDB Collections, is significantly enriched in the RNA‐sequencing dataset of NCoA6‐NC/sh PANC‐1 cells. (G) The main enriched GO/KEGG results of NCoA6‐NC/sh PANC‐1 cells.

Article Snippet: Incubation with primary antibodies for NCoA6 (1:100, Proteintech) and FBW7 (1:200, Bethyl) was at 4°C overnight, followed by polymeric HRP‐labeled anti‐rabbit IgG (Boster, China) at 37°C for 30 min. Tissue slides were scored to assess protein expression.

Techniques: RNA Sequencing, Western Blot, Negative Control, Expressing, Control, Quantitative RT-PCR

Journal: Cancer Medicine

Article Title: Nuclear receptor coactivator 6 ( NCoA6 ) promotes cell proliferation, migration, and invasion in pancreatic cancer

doi: 10.1002/cam4.6427

Figure Lengend Snippet: NCoA6 knockdown decreases the proliferation, cell cycle, and apoptosis of pancreatic cancer cells. CCK‐8 assays (A, B) and colony formation assays (C, D) were used to test the proliferation of PDAC cells transfected with NCoA6 shRNAs. The representative images and statistical results of the cell cycle (E, F) and apoptosis (G, H) using flow cytometry. (I, J) Western blotting analysis of p21 and p27 expression in NCoA6‐silenced PANC‐1 and SW1990 cells. (K, L) Western blotting analysis of CDK4, CDK2, Cyclin D1, Cyclin E1, and Cyclin A2 expression in NCoA6‐silenced PANC‐1 and SW1990 cells.

Article Snippet: Incubation with primary antibodies for NCoA6 (1:100, Proteintech) and FBW7 (1:200, Bethyl) was at 4°C overnight, followed by polymeric HRP‐labeled anti‐rabbit IgG (Boster, China) at 37°C for 30 min. Tissue slides were scored to assess protein expression.

Techniques: Knockdown, CCK-8 Assay, Transfection, Flow Cytometry, Western Blot, Expressing

Journal: Cancer Medicine

Article Title: Nuclear receptor coactivator 6 ( NCoA6 ) promotes cell proliferation, migration, and invasion in pancreatic cancer

doi: 10.1002/cam4.6427

Figure Lengend Snippet: NCoA6 knockdown decreases the migration and invasion capacity of pancreatic cancer cells. Scratch assays (A, B) and transwell migration and invasion assays (C, D) were performed to assess the effect of NCoA6 silencing on pancreatic cancer cell lines. (E, F) Western blotting analysis of EMT protein expression in NCoA6‐silenced PANC‐1 and SW1990 cells.

Article Snippet: Incubation with primary antibodies for NCoA6 (1:100, Proteintech) and FBW7 (1:200, Bethyl) was at 4°C overnight, followed by polymeric HRP‐labeled anti‐rabbit IgG (Boster, China) at 37°C for 30 min. Tissue slides were scored to assess protein expression.

Techniques: Knockdown, Migration, Western Blot, Expressing

Journal: Cancer Medicine

Article Title: Nuclear receptor coactivator 6 ( NCoA6 ) promotes cell proliferation, migration, and invasion in pancreatic cancer

doi: 10.1002/cam4.6427

Figure Lengend Snippet: Potential endogenous interactions of NCoA6, HDAC1, FBW7, and CDX2. (A, B) Co‐immunoprecipitation validated NCoA6 interaction with HDAC1 in PDAC cells. NCoA6 expression was negatively correlated with FBW7 and CDX2 expression. (C) Western blotting validated NCoA6 downregulation in FBW7 overexpressing PANC‐1 and SW1990 cells. (D, E) Correlation analysis of NCoA6 expression and FBW7 expression in PDAC tissues, as determined by the IHC score (*** p < 0.001). (F, G) Western blotting and RT‐qPCR validated CDX2 upregulation in NCoA6 knockdown PDAC cells. (H, I) Western blotting and RT‐qPCR validated CDX2 upregulation in HDAC1 knockdown PDAC cells. (J) AcH3, AcH4, and CDX2 were upregulated in HDAC inhibitor TSA‐treated PDAC cells (western blotting).

Article Snippet: Incubation with primary antibodies for NCoA6 (1:100, Proteintech) and FBW7 (1:200, Bethyl) was at 4°C overnight, followed by polymeric HRP‐labeled anti‐rabbit IgG (Boster, China) at 37°C for 30 min. Tissue slides were scored to assess protein expression.

Techniques: Immunoprecipitation, Expressing, Western Blot, Quantitative RT-PCR, Knockdown

Journal: Cancer Medicine

Article Title: Nuclear receptor coactivator 6 ( NCoA6 ) promotes cell proliferation, migration, and invasion in pancreatic cancer

doi: 10.1002/cam4.6427

Figure Lengend Snippet: Schematic representation of the working model designed using FigDraw. NCoA6 plays an important role in promoting PDAC cell proliferation, migration, invasion and progression, and is potentially related to the expression of HDAC1, FBW7, and CDX2.

Article Snippet: Incubation with primary antibodies for NCoA6 (1:100, Proteintech) and FBW7 (1:200, Bethyl) was at 4°C overnight, followed by polymeric HRP‐labeled anti‐rabbit IgG (Boster, China) at 37°C for 30 min. Tissue slides were scored to assess protein expression.

Techniques: Migration, Expressing