Journal: Cancer Science

Article Title: Downregulation of 15‐hydroxyprostaglandin dehydrogenase by interleukin‐1β from activated macrophages leads to poor prognosis in pancreatic cancer

doi: 10.1111/cas.13467

Figure Lengend Snippet: Low 15‐hydroxyprostaglandin dehydrogenase (15‐ PGDH ) expression is implicated in poor pancreatic ductal adenocarcinoma ( PDAC ) prognosis. A, Workflow diagram of patients who underwent pancreatic resection and contributed samples for immunohistochemical ( IHC ) analysis. B, Representative IHC staining of 15‐ PGDH expression in 107 PDAC tissues. Scale bar = 100 μm. C,D, Relationship between 15‐ PGDH expression and relapse‐free survival (C) or overall survival (D) using the Kaplan‐Meier method

Article Snippet: The human PDAC cell lines PK‐8 and S2‐013 were obtained from the Japanese Collection of Research Bioresource Cell Bank (Ibaraki, Japan) and RIKEN Bioresource Center Cell Bank (Tsukuba, Japan).

Techniques: Expressing, Immunohistochemical staining, Immunohistochemistry

Journal: Cancer Science

Article Title: Downregulation of 15‐hydroxyprostaglandin dehydrogenase by interleukin‐1β from activated macrophages leads to poor prognosis in pancreatic cancer

doi: 10.1111/cas.13467

Figure Lengend Snippet: 15‐Hydroxyprostaglandin dehydrogenase (15‐ PGDH ) downregulation by interleukin‐1β ( IL ‐1β) enhances pancreatic ductal adenocarcinoma cell growth. A,B, Expression of HPGD (the gene coding 15‐ PGDH protein, upper panel) or 15‐ PGDH (lower panel) in PK ‐8 cells (A) or S2‐013 cells (B) after treatment with si RNA targeting 15‐ PGDH or with control si RNA , evaluated by quantitative RT ‐ PCR (upper panel) or Western blot analysis (lower panel). Data are presented as the treated/control cell ratio. C,D, PK ‐8 cells (C) or S2‐013 cells (D) transfected with si RNA s targeting 15‐ PGDH or with control si RNA were incubated for up to 96 hours and assayed for cell number; data are presented as the treated/control (time = 0) cell ratio. E,F, Expression of 15‐ PGDH in PK ‐8 cells or S2‐013 cells after IL ‐1β (E) or tumor necrosis factor‐α ( TNF ‐α) (F) treatment for 24 and 48 hours and distilled water treatment for 48 hours as a control was evaluated by Western blotting. G, Column graph showing relative 15‐ PGDH levels in PK ‐8 cells or S2‐013 cells after IL ‐1β and TNF ‐α treatment for 24 and 48 hours, and distilled water treatment for 48 hours as a control, were evaluated using ImageJ software. H, Expression of HPGD and IL 1B in six PDAC patients determined by quantitative RT ‐ PCR . Data were normalized to the ACTB mRNA level and are shown as the mean ± SD of three independent experiments. **P < .01

Article Snippet: The human PDAC cell lines PK‐8 and S2‐013 were obtained from the Japanese Collection of Research Bioresource Cell Bank (Ibaraki, Japan) and RIKEN Bioresource Center Cell Bank (Tsukuba, Japan).

Techniques: Expressing, Control, Quantitative RT-PCR, Western Blot, Transfection, Incubation, Software

Journal: Cancer Science

Article Title: Downregulation of 15‐hydroxyprostaglandin dehydrogenase by interleukin‐1β from activated macrophages leads to poor prognosis in pancreatic cancer

doi: 10.1111/cas.13467

Figure Lengend Snippet: Tumor‐associated macrophages are inversely correlated with pancreatic ductal adenocarcinoma ( PDAC ) cells harboring high 15‐hydroxyprostaglandin dehydrogenase (15‐ PGDH ) expression. A,B, Representative immunohistochemical ( IHC ) staining of 15‐ PGDH (upper panel) and CD 163 (lower panel) expression in high 15‐ PGDH (A) and low 15‐ PGDH (B) serial PDAC specimens. Scale bar = 200 μm. C, Graph showing Pearson's correlation between the expression of 15‐ PGDH and the number of CD 163‐positive cells in 107 PDAC patients. D, Schematic representation of the findings of this study. IL ‐1βR, interleukin‐1β receptor

Article Snippet: The human PDAC cell lines PK‐8 and S2‐013 were obtained from the Japanese Collection of Research Bioresource Cell Bank (Ibaraki, Japan) and RIKEN Bioresource Center Cell Bank (Tsukuba, Japan).

Techniques: Expressing, Immunohistochemical staining, Immunohistochemistry

Journal: International journal of molecular sciences

Article Title: Complexing the Oncolytic Adenoviruses Ad∆∆ and Ad-3∆-A20T with Cationic Nanoparticles Enhances Viral Infection and Spread in Prostate and Pancreatic Cancer Models.

doi: 10.3390/ijms23168884

Figure Lengend Snippet: Figure 1. Cationic AuNP combined with Ad5wtGFP enhanced viral uptake in PCa and PDAC cell lines. (A) PCa cells were infected with Ad5wtGFP at 10, 100 and 500 ppc (PC3) and 10 and 50 ppc (22Rv) in the presence or absence of 0.5 pmol AuNPs. (B) PDAC cells were infected with Ad5wtGFP at 100 ppc and the stellate PS1 cells at 500 ppc in the presence or absence of 0.2 pmol AuNPs; samples were analysed 24 and 48 h after infection. (A,B) The EGFP signal was measured as an indication of infection levels by flow cytometry, averages ±SD, n = 3, ** p < 0.01, *** p < 0.001. (C) Fluorescent images of Panc04.03, PT45 and PS1 cells 24 and 48 h after infection with Ad5wtGFP at 50 ppc with and without AuNP at 0.2 pmol, representative of 3 studies. Images were taken at 10× magnification (Olympus IX70 microscope, Evident Europe GmbH, Stansted, UK).

Article Snippet: Human prostate cancer PC3 and 22Rv1 (ATCC, LGC Standards, UK), and human pancreatic ductal adenocarcinoma (PDAC) Panc04.03 (ATCC) and PT45 (Prof H. Kalthoff, Comprehensive Cancer Centre, Campus Kiel, Kiel, Germany) cell lines were used in the study.

Techniques: Infection, Cytometry, Microscopy

Journal: International journal of molecular sciences

Article Title: Complexing the Oncolytic Adenoviruses Ad∆∆ and Ad-3∆-A20T with Cationic Nanoparticles Enhances Viral Infection and Spread in Prostate and Pancreatic Cancer Models.

doi: 10.3390/ijms23168884

Figure Lengend Snippet: Figure 2. Ad∆∆-induced cell killing is enhanced by AuNPs in PCa and PDAC cells. (A) Dose–response to Ad∆∆in PC3 and 22Rv cells with and without 0.1 pmol AuNPs. Cell viability determined by MTS assay. (B) EC50-values (left panel) and the relative decreases in EC50-values (right panel) in each cell line compared to Ad∆∆alone. (C) PC3 cells treated with fixed doses of Ad∆∆at 500 ppc or 750 ppc and/or mitoxantrone (450 nM) with and without AuNPs at 0.1 pmol. Cell viability was measured by MTS assay 4d after infection, ** p < 0.01 (Ad/AuNP vs. Ad) and $$ p < 0.01 (Ad/M vs. Ad). (D) EC50-values generated from Ad∆∆dose–response curves ± AuNPs at 0.2 pmol (left panel) and the corresponding relative decreases in Panc04.03, PT45 and PS1 cells compared to Ad∆∆alone. (A–D) Averages ± SD, n = 3, * p < 0.05, ** p < 0.01.

Article Snippet: Human prostate cancer PC3 and 22Rv1 (ATCC, LGC Standards, UK), and human pancreatic ductal adenocarcinoma (PDAC) Panc04.03 (ATCC) and PT45 (Prof H. Kalthoff, Comprehensive Cancer Centre, Campus Kiel, Kiel, Germany) cell lines were used in the study.

Techniques: MTS Assay, Infection, Generated

Journal: International journal of molecular sciences

Article Title: Complexing the Oncolytic Adenoviruses Ad∆∆ and Ad-3∆-A20T with Cationic Nanoparticles Enhances Viral Infection and Spread in Prostate and Pancreatic Cancer Models.

doi: 10.3390/ijms23168884

Figure Lengend Snippet: Figure 3. Ad∆∆replication is enhanced in the presence of AuNP and efficiently eliminates PDAC cells in three-dimensional co-cultures with PS1 stellate cells. (A) PC3 and 22 Rv cells were infected with Ad∆∆at 100 ppc in the presence or absence of 0.1 pmol AuNPs. PC3 cells were also treated with mitoxantrone (450 nM) and infected with Ad∆∆with and without preincubation with 0.5 pmol AuNP. (B) Panc04.03 and PT45 cells were infected at 100 pc and PS1 cells at 500 ppc with and without AuNPs at 0.2 pmol and analysed 48 h after infection. (A,B) Viral replication was determined by TCID50 assays 48 and 72 h (Pca cells) and 48 h (PDAC cells) after infection, averages ± SD, n = 3, * p < 0.05, ** p < 0.01. (C) Co-cultures of Panc04.03:PS1 (1:2) cells infected with Ad5wt (1000 or 2000 ppc) in the presence or absence of AuNP (0.5 pmol), H/E staining. Cells were cultured for 3d prior to infection with virus ± AuNPs, fixed and processed for IHC 5d post-infection, 8d-old cultures, 10× magnification, representative of three biological repeats. (D) Confocal images of co- cultures detailed in (C). Localisation of virus detected by GFP-labelled secondary antibody to the E1A-antibody (green) and nuclear DAPI stain (blue) (10× magnification).

Article Snippet: Human prostate cancer PC3 and 22Rv1 (ATCC, LGC Standards, UK), and human pancreatic ductal adenocarcinoma (PDAC) Panc04.03 (ATCC) and PT45 (Prof H. Kalthoff, Comprehensive Cancer Centre, Campus Kiel, Kiel, Germany) cell lines were used in the study.

Techniques: Infection, Staining, Cell Culture, Virus

Journal: International journal of molecular sciences

Article Title: Complexing the Oncolytic Adenoviruses Ad∆∆ and Ad-3∆-A20T with Cationic Nanoparticles Enhances Viral Infection and Spread in Prostate and Pancreatic Cancer Models.

doi: 10.3390/ijms23168884

Figure Lengend Snippet: Figure 4. Ad-3∆-A20T infection and replication is enhanced in the presence of AuNP in PDAC cells. (A) Panc04.03 and PT45 were infected with Ad-3∆-A20T-GFP (100 ppc) and PS1 (500 ppc) in the presence or absence of 0.2 pmol AuNPs; samples were analysed 24 and 48 h after infection by flow cytometry for GFP expression (left panel), averages ± SD, n = 3, ** p < 0.01, *** p < 0.001. Lower panel: representative fluorescent images of PS1 cells 24 h and 48 h after infection as in (A), images were taken at 10× magnification (Olympus IX70 microscope). (B) Replication rate in Panc04.03, PT45 and PS1 cells, determined by qPCR for viral genome copies. Cells were infected as above with Ad∆∆and Ad-3∆-A20T in the presence or absence of AuNPs at 0.2 pmol and viral DNA quantified after 24, 48 and 72 h. Data presented from one experiment in triplicates relative to the respective virus alone at 24 h, averages ± SEM.

Article Snippet: Human prostate cancer PC3 and 22Rv1 (ATCC, LGC Standards, UK), and human pancreatic ductal adenocarcinoma (PDAC) Panc04.03 (ATCC) and PT45 (Prof H. Kalthoff, Comprehensive Cancer Centre, Campus Kiel, Kiel, Germany) cell lines were used in the study.

Techniques: Infection, Cytometry, Expressing, Microscopy, Virus

Journal: Journal of translational medicine

Article Title: A novel complement-fixing IgM antibody targeting GPC1 as a useful immunotherapeutic strategy for the treatment of pancreatic ductal adenocarcinoma.

doi: 10.1186/s12967-023-04745-9

Figure Lengend Snippet: Fig. 1 In-vitro characterization of AT101. A Schematic representation of GPC1 protein and visualization of the aminoacids employed for the mouse immunization. B SDS-PAGE analysis performed in non-reducing and reducing condition to determine the IgM nature of AT101. C Elisa to assess the GPC1 binding activity of AT101, the positive control (CTRL +) was represented by serum of mice immunized with GPC1, the negative control (CTRL-) was performed by only using the goat anti-mouse IgG/IgM antibody conjugated with alkaline phosphatase (AP) without AT101 (n = 3). D Flow cytometry analysis to evaluate GPC1 expression in BXPC3 and Jurkat cells using AT101 and the commercial anti-GPC1 antibody as positive control (n = 3). ***, 0.001 < p ≤ 0.0001

Article Snippet: BXPC3 cell line (human pancreatic ductal adenocarcinoma) (RRID: CVCL_0186) was purchased from ATCC and cultured in DMEM.

Techniques: In Vitro, SDS Page, Enzyme-linked Immunosorbent Assay, Binding Assay, Activity Assay, Positive Control, Negative Control, Flow Cytometry, Expressing