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: Nature Communications

Article Title: Tumour-derived Interleukin 35 promotes pancreatic ductal adenocarcinoma cell extravasation and metastasis by inducing ICAM1 expression

doi: 10.1038/ncomms14035

Figure Lengend Snippet: Consecutive sections of pancreatic carcinoma tissues were used to analyse the expression levels of the two subunits of the IL-35 ligand: EBI3 and P35. ( a ) IHC staining of EBI3 and P35 proteins in five representative histologic types of pancreatic carcinoma (PDAC, mucinous adenocarcinoma, giant cell carcinoma, malignant neuroendocrine carcinoma and mucous cell carcinoma) and the adjacent normal pancreatic tissues. ( b ) Representative images are shown for absent, low, median and high expression of EBI3 and P35 in IHC staining in PDAC tissues. ( c – e ) The distribution of IHC results ( c ), staining extent correlation ( d , Pearson's correlation analysis) and IHC result correlation ( e , Spearman's correlation analysis) analysis of EBI3 and P35 in 123 PDAC patients. ( f ) Spearman's correlation analysis of the mRNA expression profiles of EBI3 and P35 in 157 PDAC patients from TCGA. ( g ) Co-immunoprecipitation and western blot analysis of EBI3 and P35 proteins from tumour tissue lysates of three PDAC patients, the remaining supernatant post immunoprecipitation (Post-IP) was also detected. ( h , i ) Kaplan–Meier analysis of OS ( h ) and RFS ( i ) of 123 PDAC patients ( P =0.0010 and 0.0072, respectively, by log-rank test) according to different IL-35 levels. The test of significance is two-sided. Figure panel pairs in a and b represent images taken with different zooming options; scale bars, 100 μm.

Article Snippet: The primary PDAC cell lines PTX0001, PTX0015, PTX0037 and PTX 0049 were purchased from WuXi Pharmatech Company (WuXi, China).

Techniques: Expressing, Immunohistochemistry, Staining, Immunoprecipitation, Western Blot

Journal: Nature Communications

Article Title: Tumour-derived Interleukin 35 promotes pancreatic ductal adenocarcinoma cell extravasation and metastasis by inducing ICAM1 expression

doi: 10.1038/ncomms14035

Figure Lengend Snippet: ( a ) Spearman's correlation analysis of the mRNA expression profiles of EBI3 and ICAM1 (left), as well as P35 and ICAM1 (right), in 157 PDAC patients from TCGA. ( b ) The 157 patients were stratified into two groups on the basis of the mRNA level of EBI3 and P35 : IL-35 high, both EBI3 and P35 mRNA levels higher than median; others were IL-35 low. The ICAM1 mRNA levels of the two groups are presented. Unpaired t -tests were used; shown are the mean values±s.d. ( c ) Representative IHC staining images of high and low levels for ICAM1, EBI3 and P35 in consecutive sections are shown. Figure panels pairs represents images taken with different zooming options; scale bars, 100 μm. ( d ) Real distribution of IHC results between ICAM1 and IL-35 expression. ( e ) Spearman's correlation analysis of ICAM1 and IL-35 IHC staining results in 123 human PDAC surgical samples. ( f ) Real distribution of immunohistochemical results of ICAM1 in 123 PDAC tissues. ( g ) Kaplan–Meier analysis of OS of 123 PDAC patients according to different ICAM1 levels by IHC staining (ICAM1 low, −/+; ICAM1 high, ++/+++). ( h ) mRNA profiles and follow-up data of 130 PDAC patients from TCGA analysed for the correlation of the ICAM1 mRNA expression and survival. Log-rank tests were used in g and h . Tests of significance are two-sided; * P <0.05.

Article Snippet: The primary PDAC cell lines PTX0001, PTX0015, PTX0037 and PTX 0049 were purchased from WuXi Pharmatech Company (WuXi, China).

Techniques: Expressing, Immunohistochemistry, Immunohistochemical staining

Journal: Nature Communications

Article Title: Tumour-derived Interleukin 35 promotes pancreatic ductal adenocarcinoma cell extravasation and metastasis by inducing ICAM1 expression

doi: 10.1038/ncomms14035

Figure Lengend Snippet: ( a ) Examples are shown for absent, low, median and high expressions of GP130 and IL-12Rβ2 in PDAC tissues. Figure panel pairs represents images taken with different zooming options; scale bars, 100 μm. ( b ) Real distribution of IHC results of GP130 and IL-12Rβ2. ( c ) Examples are shown for the co-expression of IL-35 and IL-35R in PDAC tissues; scale bars, 100 μm. ( d ) Kaplan–Meier analysis of OS (left) and RFS (right) of 123 PDAC patients according to the expression levels of IL-35 and GP130. ( e ) Kaplan–Meier analysis as in d but according to the expression levels of IL-35 and IL-12Rβ2. Log-rank tests were used in d and e .

Article Snippet: The primary PDAC cell lines PTX0001, PTX0015, PTX0037 and PTX 0049 were purchased from WuXi Pharmatech Company (WuXi, China).

Techniques: Expressing

Journal: Nature Communications

Article Title: Tumour-derived Interleukin 35 promotes pancreatic ductal adenocarcinoma cell extravasation and metastasis by inducing ICAM1 expression

doi: 10.1038/ncomms14035

Figure Lengend Snippet: ( a , b ) DiI-labelled PDAC cells were injected into green fluorescent protein-knockin mice through the portal veins. At the time points of 6 and 24 h, animals were killed and livers were sliced. ( a ) Representative images of adhered/arrested tumour cells (white arrow) and extravasated tumour cells (red arrow) in livers are shown ( n =6 for each group). ( b ) Numbers of adhered/arrested tumour cells and extravasated cells are analysed. ( c – f ) Indicated tumour cells were orthotopically transplanted into the SCID mouse pancreases to develop tumours ( n =8 for each group). Representative images of the tumours and metastatic sites of livers are shown in c . Hematoxylin and eosin staining images of liver micrometastasis sites are shown in d . The total numbers of visible metastatic lesions in c and invisible micrometastases in d are presented in e and f , respectively. ( g ) Schematic diagram of pancreatic carcinoma undergoing a metastatic progression mediated by the IL-35-ICAM1 axis. After being secreted by pancreatic tumour cells, IL-35 acts on the tumour cells in an autocrine or paracrine manner. After binding to the homodimer GP130-GP130, IL-35 activates the Jak-STAT signalling pathway. STAT1 is phosphorylated and forms the p-STAT1/p-STAT1 homodimer. Then, the homodimers are transferred into the nucleus, where they activate the transcription of the ICAM1 gene. The ICAM1 protein on the tumour cell surface enhances the extravasation process by facilitating adhesion to the endothelial cells and the transendothelial migration process in an ICAM1–fibrinogen–ICAM1 manner. The elevated extravasation activity promotes metastasis in distant organs or tissues. Unpaired t -tests were used in b , e and f ; tests of significance are two-sided; shown are the mean values±s.d.; * P <0.05. Figure panel pairs in a represents images taken with different zooming options; scale bar, 100 μm ( a ) and 50 μm ( d ).

Article Snippet: The primary PDAC cell lines PTX0001, PTX0015, PTX0037 and PTX 0049 were purchased from WuXi Pharmatech Company (WuXi, China).

Techniques: Injection, Knock-In, Staining, Binding Assay, Migration, Activity Assay

Journal: Oncotarget

Article Title: Perspectives of TGF-β inhibition in pancreatic and hepatocellular carcinomas

doi:

Figure Lengend Snippet: At the cellular level, TGF-β induces proliferation and survival of PDAC cells in the late phase of PDAC carcinogenesis (after SMAD4 inactivation), and promotes epithelial-to-mesenchymal transition (EMT), invasion, and metastasis. At the microenvironment level, TGF-β is a key mediator of the dialogue between cancer and stellate cells (fibrotic cells), involved in the production of a dense fibrotic stroma and the resulting low vascularization of PDAC. TGF-β also deregulates the immune microenvironment toward immunosuppression and inappropriate inflammation.

Article Snippet: Consistent with this, Bachem et al.[ ] showed that PDAC cell lines stimulated PSC proliferation.

Techniques:

Journal: Journal of Gastroenterology

Article Title: Increased expression of PHD3 represses the HIF-1 signaling pathway and contributes to poor neovascularization in pancreatic ductal adenocarcinoma

doi: 10.1007/s00535-014-1030-3

Figure Lengend Snippet: Immunohistological staining for the expressions of CD105 and CD44 in surgical specimens from PDAC and HCC patients. a The expression of CD105, a marker for neovascularization, was extremely low in the PDAC specimens compared to the HCC specimens. The quantitative data and results of the statistical analysis between the groups are shown in the lower bar graph. There was a significant difference with the vascularity between PDAC and HCC. b A higher expression of CD44, a marker for cancer stem cells was observed in the PDAC specimens compared to the HCC specimens. The quantitative data and results of the statistical analysis between the groups are shown in the lower bar graph. c There was a significant negative correlation between the expressions of CD105 and CD44 in each specimens, for both PDAC ( p < 0.01, left ) and HCC ( p < 0.001, right )

Article Snippet: Two PDAC lines (HS766T and Panc-1) and two HCC cell lines (HepG2 and HuH7) purchased from the Japanese Collection of Research Bioresources (Osaka, Japan) were used for the experiments.

Techniques: Staining, Expressing, Marker

Journal: Journal of Gastroenterology

Article Title: Increased expression of PHD3 represses the HIF-1 signaling pathway and contributes to poor neovascularization in pancreatic ductal adenocarcinoma

doi: 10.1007/s00535-014-1030-3

Figure Lengend Snippet: Immunohistological staining for the expressions of HIF-1α, PHD3, and Siah2 in surgical specimens from PDAC and HCC patients. a HIF-1α was positively stained in the nuclei of cancer cells in HCC specimens, but was negatively or extremely rarely stained in the PDAC specimens, and a semiquantitative analysis showed a significant difference between the groups ( p < 0.05). b PHD3 was positively stained in many cancer cells in the PDAC specimens, but was negatively or weakly stained in the HCC specimens ( p < 0.001). c The expression of Siah2 was negative or weak in both the PDAC and HCC specimens, but the semiquantitative analysis showed a significantly higher expression in the HCC specimens ( p < 0.05)

Article Snippet: Two PDAC lines (HS766T and Panc-1) and two HCC cell lines (HepG2 and HuH7) purchased from the Japanese Collection of Research Bioresources (Osaka, Japan) were used for the experiments.

Techniques: Staining, Expressing

Journal: Journal of Gastroenterology

Article Title: Increased expression of PHD3 represses the HIF-1 signaling pathway and contributes to poor neovascularization in pancreatic ductal adenocarcinoma

doi: 10.1007/s00535-014-1030-3

Figure Lengend Snippet: The expression of CD44 in PDAC (HS766T and Panc-1) and HCC (HepG2 and HuH7) cell lines. Cells were cultured in 20 % O 2 or 1 % O 2 for 24 h, and the expression of the CSC marker CD44 was then estimated by flow cytometry and compared to cancer stem cell markers by hypoxic stimulation. The expression of CD44 was much higher in the PDAC cell lines compared to the HCC cell lines, but the 24-h incubation in 1 % O 2 did not change the expression of CD44 in any of the cell lines. The green line shows the expression of CD44 in hypoxia (1 % O 2 for 24 h) and the purple area shows it in normoxia (20 % O 2 for 24 h)

Article Snippet: Two PDAC lines (HS766T and Panc-1) and two HCC cell lines (HepG2 and HuH7) purchased from the Japanese Collection of Research Bioresources (Osaka, Japan) were used for the experiments.

Techniques: Expressing, Cell Culture, Marker, Flow Cytometry, Incubation

Journal: Journal of Gastroenterology

Article Title: Increased expression of PHD3 represses the HIF-1 signaling pathway and contributes to poor neovascularization in pancreatic ductal adenocarcinoma

doi: 10.1007/s00535-014-1030-3

Figure Lengend Snippet: Western-blot analysis of the expressions of PHD3 and HIF-1α in the PDAC (HS766T and Panc-1) and HCC (HepG2 and HuH7) cell lines. a The expression of PHD3 was clearly detected in both of the PDAC cell lines, but negatively or very dimly in the two HCC cell lines cultured in the 20 or 1 % O 2 conditions. Although comparable expression levels of HIF-1α were detected between the PDAC and HCC cell lines under 20 % O 2 , the hypoxic stimulation by 1 % O 2 significantly enhanced the expression of HIF-1α only in the two HCC cell lines, but only slightly upregulated the expression in the PDAC cell lines. b The semiquantitative data are shown

Article Snippet: Two PDAC lines (HS766T and Panc-1) and two HCC cell lines (HepG2 and HuH7) purchased from the Japanese Collection of Research Bioresources (Osaka, Japan) were used for the experiments.

Techniques: Western Blot, Expressing, Cell Culture

Journal: Journal of Gastroenterology

Article Title: Increased expression of PHD3 represses the HIF-1 signaling pathway and contributes to poor neovascularization in pancreatic ductal adenocarcinoma

doi: 10.1007/s00535-014-1030-3

Figure Lengend Snippet: VEGF secretion from the PDAC (HS766T and Panc-1) and HCC (HepG2 and HuH7) cell lines. The level of VEGF in conditioned medium was measured by ELISA. The secretion of VEGF was comparable between the PDAC and HCC cell lines under 20 % O 2 . Hypoxic stimulation by 1 % O 2 significantly enhanced the VEGF secretion from both of the HCC cell lines ( p < 0.05), but no significant enhancement was observed in the PDAC cell lines

Article Snippet: Two PDAC lines (HS766T and Panc-1) and two HCC cell lines (HepG2 and HuH7) purchased from the Japanese Collection of Research Bioresources (Osaka, Japan) were used for the experiments.

Techniques: Enzyme-linked Immunosorbent Assay

Journal: Journal of Gastroenterology

Article Title: Increased expression of PHD3 represses the HIF-1 signaling pathway and contributes to poor neovascularization in pancreatic ductal adenocarcinoma

doi: 10.1007/s00535-014-1030-3

Figure Lengend Snippet: Knockdown of PHD3 in the PDAC cell line HS766T. a Knockdown of PHD3 was done by siRNA, and the expression levels of PHD3 and HIF-1α were estimated by Western blot. PHD3 knockdown did not change the expression of HIF-1α when the cells were cultured in 20 % O 2 , but the expression of HIF-1α was clearly enhanced when the cells were cultured in 1 % O 2 . b The semiquantitative data in HIF-1α are shown. c Hypoxic stimulation by 1 % O 2 significantly enhanced the VEGF secretion from HS766T cells treated by siPHD3 ( p < 0.05)

Article Snippet: Two PDAC lines (HS766T and Panc-1) and two HCC cell lines (HepG2 and HuH7) purchased from the Japanese Collection of Research Bioresources (Osaka, Japan) were used for the experiments.

Techniques: Knockdown, Expressing, Western Blot, Cell Culture