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: Clinical & experimental metastasis

Article Title: Genome-wide in vivo RNAi screen identifies ITIH5 as a metastasis suppressor in pancreatic cancer

doi: 10.1007/s10585-017-9840-3

Figure Lengend Snippet: A, Schematic diagram of the in vivo shRNA forward screen to identify genes regulating liver metastasis in PDAC. Briefly, following transduction, S2-028 PDAC cells were injection intrasplenically. After 4 wk, liver metastases were isolated, cultured and re-injected into the spleens of athymic mice. Hepatic metastases were isolated and established in cell culture before isolating DNA and identification of shRNA integrated. After the second round of intrasplenic injections, two target genes were identified from 2 cell clones, HMP19 and ITIH5. B, Representative images of livers from mice injected with control S2-028 or shRNA library-infected S2-028. Scale bar, 10 mm.

Article Snippet: Cell lines and cell culture Five human PDAC cell lines (S2-007, S2-028, MIAPaCa-2, BxPC-3 and Panc-1) were obtained from Dr. M. Anthony Hollingsworth (Eppley Cancer Center).

Techniques: In Vivo, shRNA, Transduction, Injection, Isolation, Cell Culture, Clone Assay, Control, Infection

Journal: Clinical & experimental metastasis

Article Title: Genome-wide in vivo RNAi screen identifies ITIH5 as a metastasis suppressor in pancreatic cancer

doi: 10.1007/s10585-017-9840-3

Figure Lengend Snippet: A, The relative expression ITIH5 levels of in a panel of PDAC cell lines and HPNE, as determined by immunoblotting. Relative metastatic potential is provided for comparison. B, C Mixed pools of S2-007 and MIAPaCa-2 cells transduced with ITIH5 cDNA over-express ITIH5 and the number of liver metastasis are compared following intrasplenic injections. Representative livers are shown. Arrows highlight liver foci. Results are means + SE of liver nodules from positive mice. * P < 0.05 and ** P < 0.001; Mann-Whitney U test.

Article Snippet: Cell lines and cell culture Five human PDAC cell lines (S2-007, S2-028, MIAPaCa-2, BxPC-3 and Panc-1) were obtained from Dr. M. Anthony Hollingsworth (Eppley Cancer Center).

Techniques: Expressing, Western Blot, Comparison, Transduction, MANN-WHITNEY

Journal: Clinical & experimental metastasis

Article Title: Genome-wide in vivo RNAi screen identifies ITIH5 as a metastasis suppressor in pancreatic cancer

doi: 10.1007/s10585-017-9840-3

Figure Lengend Snippet: A-C, Images of pancreatic tumors and average tumor volume following orthotopic injections of PDAC cells in which ITIH5 expression levels have been altered by transduction of shRNA (in S2-028 cells) or cDNA (in S2-007 or MIAPaCa-2 cells). Graphical representation of orthotopic tumor size at time of euthanasia. Results are means + SE of the orthotopic tumor volume. * P < 0.05 and ** P < 0.01; Mann-Whitney U test.

Article Snippet: Cell lines and cell culture Five human PDAC cell lines (S2-007, S2-028, MIAPaCa-2, BxPC-3 and Panc-1) were obtained from Dr. M. Anthony Hollingsworth (Eppley Cancer Center).

Techniques: Expressing, Transduction, shRNA, MANN-WHITNEY

Journal: Clinical & experimental metastasis

Article Title: Genome-wide in vivo RNAi screen identifies ITIH5 as a metastasis suppressor in pancreatic cancer

doi: 10.1007/s10585-017-9840-3

Figure Lengend Snippet: Representative photographs showing migration (A-C) and invasion (D-F) are inhibited when ITIH5 levels are high. Results are means + SE of the % wound closure and number of invaded cells, respectively. G-I, Growth curves of PDAC cell lines altered ITIH5 expression levels. * P < 0.01 and ** P < 0.001; Mann-Whitney U test. Scale bar, 200 μm.

Article Snippet: Cell lines and cell culture Five human PDAC cell lines (S2-007, S2-028, MIAPaCa-2, BxPC-3 and Panc-1) were obtained from Dr. M. Anthony Hollingsworth (Eppley Cancer Center).

Techniques: Migration, Expressing, MANN-WHITNEY