Journal: International Journal of Molecular Medicine

Article Title: FUBP1 mediates the growth and metastasis through TGFβ/Smad signaling in pancreatic adenocarcinoma

doi: 10.3892/ijmm.2021.4899

Figure Lengend Snippet: Upregulated FUBP1 expression levels are associated with a poor prognosis in patients with PAAD. (A) FUBP1 expression levels in PAAD and adjacent normal tissues from The Cancer Genome Atlas database. (B) starBase database was used to determine that, compared with patients with PAAD with low expression levels of FUBP1, patients with high expression levels of FUBP1 had a poorer overall survival (P=0.037). (C) mRNA expression levels of FUBP1 were upregulated in seven human PAAD tissues compared with adjacent normal tissues. Data are presented as the mean ± SD from three independent experiments. * P<0.05 and *** P<0.01. FUBP1, far upstream element-binding protein 1; PAAD, pancreatic adenocarcinoma.

Article Snippet: Human PAAD cell lines (BxPC-3, PaTu8988, PANC-1 and SW1990) were conserved at the Central Laboratory of Shanghai Fengxian District Central Hospital, and were cultured in DMEM supplemented with 10% FBS (both from Gibco; Thermo Fisher Scientific, Inc.), and maintained in a humidified incubator with 5% CO 2 at 37°C.

Techniques: Expressing, Binding Assay

Journal: International Journal of Molecular Medicine

Article Title: FUBP1 mediates the growth and metastasis through TGFβ/Smad signaling in pancreatic adenocarcinoma

doi: 10.3892/ijmm.2021.4899

Figure Lengend Snippet: Expression levels of FUBP1 in PAAD cell lines. (A) mRNA and protein expression levels of FUBP1 in four human PAAD cell lines. (B) Transfection efficiency of si-FUBP1 or si-NC in PaTu8988 cells was analyzed using RT-qPCR and western blotting. (C) Transfection efficiency of FUBP1-OE or empty vector in SW1990 cells was analyzed using RT-qPCR and western blotting. Data are presented as the mean ± SD from three independent experiments. * P<0.05. FUBP1, far upstream element-binding protein 1; PAAD, pancreatic adenocarcinoma; si, small interfering RNA; NC, negative control; OE, overexpression; RT-qPCR, reverse transcription-quantitative PCR.

Article Snippet: Human PAAD cell lines (BxPC-3, PaTu8988, PANC-1 and SW1990) were conserved at the Central Laboratory of Shanghai Fengxian District Central Hospital, and were cultured in DMEM supplemented with 10% FBS (both from Gibco; Thermo Fisher Scientific, Inc.), and maintained in a humidified incubator with 5% CO 2 at 37°C.

Techniques: Expressing, Transfection, Quantitative RT-PCR, Western Blot, Plasmid Preparation, Binding Assay, Small Interfering RNA, Negative Control, Over Expression, Reverse Transcription, Real-time Polymerase Chain Reaction

Journal: Cancer Science

Article Title: Deubiquitinase ubiquitin‐specific peptidase 10 maintains cysteine rich angiogenic inducer 61 expression via Yes1 associated transcriptional regulator to augment immune escape and metastasis of pancreatic adenocarcinoma

doi: 10.1111/cas.15326

Figure Lengend Snippet: Ubiquitin‐specific peptidase 10 (USP10) is abundantly expressed in pancreatic adenocarcinoma (PAAD) tissues signaling pathway enriched on the positive side of the YAP1 according to Gene Set Enrichment Analysis (GSEA and cells and positively correlated with YAP1. (A) Hydrolysis‐related); (B) genes show high correlation with YAP1 analyzed using a correlation test (cor. Test); (C) USP10 expression in TCGA‐PAAD and GTEx databases; (D) association between USP10 level and the prognosis of patients analyzed in TCGA‐PAAD; (E) staining intensity of USP10 in the PAAD tissues and normal pancreatic tissues in the HPA system; (F and G) expression of Cyr61 (F) and CTGF (G) in TCGA‐PAAD; (H and I) positive correlations between CTGF (H) and Cyr61 (I), and USP10 examined by Spearman's rank correlation coefficient; (J) expression of USP10 in PAAD cell lines (Capan‐2, AsPC‐1, Panc‐1, and BxPC‐1), and in the normal HPDE6 cells determined by RT‐qPCR. Differences were analyzed by one‐way ANOVA (J), *** P < 0.001

Article Snippet: A human pancreatic duct epithelial cell line HPDE6 and PAAD cell lines (Capan‐2, Panc‐1, AsPC‐1, and BxPC‐1) were purchased from Genechem.

Techniques: Ubiquitin Proteomics, Expressing, Staining, Quantitative RT-PCR

Journal: Cancer Science

Article Title: Deubiquitinase ubiquitin‐specific peptidase 10 maintains cysteine rich angiogenic inducer 61 expression via Yes1 associated transcriptional regulator to augment immune escape and metastasis of pancreatic adenocarcinoma

doi: 10.1111/cas.15326

Figure Lengend Snippet: Ubiquitin‐specific peptidase 10 (USP10) knockdown increases ubiquitination and degradation of YAP1 in pancreatic adenocarcinoma (PAAD) cells. (A) Protein levels of YAP1, Cyr61, and USP10 in Panc‐1 and BxPC‐1 cells after shUSP10 transfection examined by western blot analysis; (B) binding relationship between USP10 and YAP1 in Panc‐1 and BxPC‐1 cells examined by the Co‐IP assay; (C) sub‐cellular distribution of USP10 and YAP1 in Panc‐1 and BxPC‐1 cells examined using dual immunofluorescence staining; (D) ubiquitination level of YAP1 in 293T cells after HA‐ubiquitin, Flag‐YAP1, or USP10 overexpression; (E) ubiquitination level of YAP1 in Panc‐1 and BxPC3 cells after USP10 knockdown examined by Co‐IP; and (F) protein level of YAP1 in Panc‐1 and BxPC3 cells after 10 μM MG132 treatment determined by western blot analysis. Differences were analyzed by two‐way ANOVA (A), ** P < 0.01

Article Snippet: A human pancreatic duct epithelial cell line HPDE6 and PAAD cell lines (Capan‐2, Panc‐1, AsPC‐1, and BxPC‐1) were purchased from Genechem.

Techniques: Ubiquitin Proteomics, Knockdown, Transfection, Western Blot, Binding Assay, Co-Immunoprecipitation Assay, Immunofluorescence, Staining, Over Expression

Journal: Cancer Science

Article Title: Deubiquitinase ubiquitin‐specific peptidase 10 maintains cysteine rich angiogenic inducer 61 expression via Yes1 associated transcriptional regulator to augment immune escape and metastasis of pancreatic adenocarcinoma

doi: 10.1111/cas.15326

Figure Lengend Snippet: Ubiquitin‐specific peptidase 10 (USP10) knockdown reduces the immune escape of pancreatic adenocarcinoma (PAAD) cells. (A and B) expression of PD‐L1 and Galectin‐9 in Panc‐1 and BxPC‐1 cells detected by RT‐qPCR and western blot analysis; (C) portion of programmed death ligand‐1 (PD‐L1)‐ and Galectin‐9‐positive Panc‐1 and BxPC‐1 cells examined by flow cytometry; (D) portion of CD86‐positive (M1‐polarized) and CD206‐positive (M2‐polarized) macrophages after co‐incubation with Panc‐1 and BxPC‐1 cells evaluated by flow cytometry; (E) expression of the M1‐macrophage markers (iNOS, IL‐1β, and IL‐6) and M2‐macrophage markers (Arg1, IL‐10, and IL‐13) in cells examined by RT‐qPCR; (F) the degree of Panc‐1 and BxPC‐1 cells lysed by natural killer (NK) cells examined using an LDH kit. Differences were analyzed by two‐way ANOVA (A–E), ** P < 0.01; *** P < 0.001

Article Snippet: A human pancreatic duct epithelial cell line HPDE6 and PAAD cell lines (Capan‐2, Panc‐1, AsPC‐1, and BxPC‐1) were purchased from Genechem.

Techniques: Ubiquitin Proteomics, Knockdown, Expressing, Quantitative RT-PCR, Western Blot, Flow Cytometry, Incubation

Journal: Cancer Science

Article Title: Deubiquitinase ubiquitin‐specific peptidase 10 maintains cysteine rich angiogenic inducer 61 expression via Yes1 associated transcriptional regulator to augment immune escape and metastasis of pancreatic adenocarcinoma

doi: 10.1111/cas.15326

Figure Lengend Snippet: Ubiquitin‐specific peptidase 10 (USP10) knockdown reduces growth and invasiveness of pancreatic adenocarcinoma (PAAD) cells in vitro . (A) viability of Panc‐1 and BxPC‐1 cells examined using a CTG kit; (B) proliferation rate of the Panc‐1 and BxPC‐1 cells using the EdU labeling kit; (C) apoptosis of Panc‐1 and BxPC‐1 cells examined using flow cytometry; (D and E) levels of EMT‐related biomarkers ZO‐1, E‐cadherin, Vimentin, and N‐cadherin in Panc‐1 and BxPC‐1 cells determined by RT‐qPCR (D), western blot analysis (E), and immunofluorescence staining (F), respectively; (G and H) migration (G) and invasion (H) ability of Panc‐1 and BxPC‐1 cells determined by Transwell assays. Differences were analyzed by two‐way ANOVA (A–E), ** P < 0.01; *** P < 0.001

Article Snippet: A human pancreatic duct epithelial cell line HPDE6 and PAAD cell lines (Capan‐2, Panc‐1, AsPC‐1, and BxPC‐1) were purchased from Genechem.

Techniques: Ubiquitin Proteomics, Knockdown, In Vitro, Labeling, Flow Cytometry, Quantitative RT-PCR, Western Blot, Immunofluorescence, Staining, Migration

Journal: Cancer Science

Article Title: Deubiquitinase ubiquitin‐specific peptidase 10 maintains cysteine rich angiogenic inducer 61 expression via Yes1 associated transcriptional regulator to augment immune escape and metastasis of pancreatic adenocarcinoma

doi: 10.1111/cas.15326

Figure Lengend Snippet: Ubiquitin‐specific peptidase 10 (USP10) knockdown suppresses growth and metastasis of pancreatic adenocarcinoma (PAAD) cells in vivo. (A) Growth rate of the xenograft tumors; (B) weight of the xenograft tumors on the 35th day; (C–G) staining intensity of KI67 (C), PCNA (D), YAP1 (E), Cyr61 (F), and CTGF (G) in the tissues of xenograft tumors detected by IHC staining; and (H and I) tumor metastasis in murine lung (H) and liver (I) tissues on the 45th day examined by HE staining. In each group, n = 6. Differences were analyzed by two‐way ANOVA (A–E), ** P < 0.01; *** P < 0.001

Article Snippet: A human pancreatic duct epithelial cell line HPDE6 and PAAD cell lines (Capan‐2, Panc‐1, AsPC‐1, and BxPC‐1) were purchased from Genechem.

Techniques: Ubiquitin Proteomics, Knockdown, In Vivo, Staining, Immunohistochemistry

Journal: Cancer Science

Article Title: Deubiquitinase ubiquitin‐specific peptidase 10 maintains cysteine rich angiogenic inducer 61 expression via Yes1 associated transcriptional regulator to augment immune escape and metastasis of pancreatic adenocarcinoma

doi: 10.1111/cas.15326

Figure Lengend Snippet: Overexpression of Cyr61 restores immune tolerance of pancreatic adenocarcinoma (PAAD) cells. (A) Protein level of Cyr61 in Panc‐1 and BxPC‐1 cells after oe‐Cyr61 transfection examined by western blot analysis; (B) PD‐L1 and Galectin‐9 expression in Panc‐1 and BxPC‐1 cells examined by RT‐qPCR; (C) portion of PD‐L1‐ and Galectin‐9‐positive cells evaluated by the flow cytometry; (D) portion of CD86‐positive (M1‐polarized) and CD206‐positive (M2‐polarized) macrophages after co‐incubation with Panc‐1 and BxPC‐1 cells detected by flow cytometry; (E) expression of the M1‐macrophage markers (iNOS, IL‐1β, and IL‐6) and M2‐macrophage markers (Arg1, IL‐10, and IL‐13) in cells examined by RT‐qPCR; (F) the degree of Panc‐1 and BxPC‐1 cells lysed by natural killer (NK) cells examined using an LDH kit. Differences were analyzed by two‐way ANOVA (A–E), ** P < 0.01

Article Snippet: A human pancreatic duct epithelial cell line HPDE6 and PAAD cell lines (Capan‐2, Panc‐1, AsPC‐1, and BxPC‐1) were purchased from Genechem.

Techniques: Over Expression, Transfection, Western Blot, Expressing, Quantitative RT-PCR, Flow Cytometry, Incubation

Journal: Cancer Science

Article Title: Deubiquitinase ubiquitin‐specific peptidase 10 maintains cysteine rich angiogenic inducer 61 expression via Yes1 associated transcriptional regulator to augment immune escape and metastasis of pancreatic adenocarcinoma

doi: 10.1111/cas.15326

Figure Lengend Snippet: Overexpression of Cyr61 restores pancreatic adenocarcinoma (PAAD) cell proliferation in vitro. (A) Viability of Panc‐1 and BxPC‐1 cells examined using a CellTiter Glo (CTG) kit; (B) proliferation rate of the Panc‐1 and BxPC‐1 cells using the EdU labeling kit; (C) apoptosis of Panc‐1 and BxPC‐1 cells examined using flow cytometry; (D–F) levels of EMT‐related biomarkers ZO‐1, E‐cadherin, Vimentin, and N‐cadherin in Panc‐1 and BxPC‐1 cells determined by RT‐qPCR (D), western blot analysis (E), and immunofluorescence staining (F), respectively; and (G and H) migration (G) and invasion (H) ability of Panc‐1 and BxPC‐1 cells determined by Transwell assays. Differences were analyzed by two‐way ANOVA (A–G), ** P < 0.01

Article Snippet: A human pancreatic duct epithelial cell line HPDE6 and PAAD cell lines (Capan‐2, Panc‐1, AsPC‐1, and BxPC‐1) were purchased from Genechem.

Techniques: Over Expression, In Vitro, Labeling, Flow Cytometry, Quantitative RT-PCR, Western Blot, Immunofluorescence, Staining, Migration