Journal: Oncotarget

Article Title: The antitumor potential of Interleukin-27 in prostate cancer.

doi: 10.18632/oncotarget.1425

Figure Lengend Snippet: Figure 1: Expression of IL-27R on hPCa cell lines and assessment of IL-27 effects on hPCa cells in vitro and in vivo. Panel A and B. Expression of WSX-1 (Panel A) and gp130 (panel B) was analysed in PC3, DU145, 22Rv1 and LNCaP cells by flow cytometry. Open profile: WSX-1 (panel A) and gp130 (panel B) staining. Dark profile: isotype matched mAb staining. Experiments were performed at least in triplicate. Panel C. Western blot analyses of WSX-1 in nuclear extracts (n.e.) and membrane extracts (m.e.) from 22Rv1 and DU145 cells. Nuclear extracts were used as negative controls. A specific 70 kDalton (Da) band, corresponding to WSX-1 protein, was observed in membrane but not nuclear extracts obtained from DU145 cells. Panel D. IL-27 inhibits PC3 cell proliferation in vitro, as assessed by CFSE staining. PC3 cells were cultured for 48 and 120 hours with medium alone (left panel) or in the presence of 100 ng/ml hrIL-27 (middle panel). Flow cytometry analyses showed that IL-27 inhibited PC3 cell proliferation after 120 hours of treatment (middle and right panel), as indicated by the higher CFSE intensity in hrIL-27 treated PC3 cells compared to untreated cells at this time point (right panel). Panel E. Flow cytometry analyses showed that IL-27 inhibited DU145 (left panel) cell proliferation after 120 hours of treatment, as indicated by the higher CFSE intensity in hrIL-27 treated cells (dark profile) compared to untreated cells at this time point (light profile). IL-27 did not affect 22Rv1 (middle panel) or LNCaP (right panel) cell proliferation at the same time point. Panel F. IL-27 did not induce apoptosis after 120 hours of treatment in PC3, DU145, 22Rv1 or LNCaP cells.

Article Snippet: Cell culture, antibodies, reagents, flow cytometry and western blot The human PC3, DU145, 22Rv1 and LNCaP PCa cell lines (LGC Standards, Teddington, UK) were cultured in RPMI 1640 with 10% FCS (Seromed-BiochromKG, Berlin, D). hrIL-27 (R&D System, Minneapolis, MN, USA) was used at 100 ng/ml following titration experiments.

Techniques: Expressing, In Vitro, In Vivo, Flow Cytometry, Staining, Western Blot, Membrane, Cell Culture

Journal: Oncotarget

Article Title: The antitumor potential of Interleukin-27 in prostate cancer.

doi: 10.18632/oncotarget.1425

Figure Lengend Snippet: Figure 2: Inhibition of human PC3 and DU145 cell growth in vivo by IL-27 treatment. Panel A. Tumor growth curve from day 14 to day 51 after PC3 s.c. cell injection into athymic nude mice was obtained by measuring tumor volume in situ using a caliper. Dark line represents growth curve (volume) of tumors formed in mice injected with PC3 cells and treated with PBS (controls). Grey light line represents growth curve of tumors formed in mice injected with PC3 cells and treated with hrIL-27. Asterisks mean significant differences (see results section). Panel B. Volume of tumor masses developed in mice injected with the DU145 cell line and subsequently treated with PBS (controls) or hrIL-27. Panel C and D. Morphological and immunohistochemical features of PC3 (C) and DU145 (D) tumor masses grown in hrIL-27 or PBS treated mice. Expression of WSX-1 was preserved in vivo by tumor developed after s.c. injection of PC3 or DU145 cells in both control (a) and hrIL-27 treated (b) mice. The proliferative activity of PC3 tumors grown in PBS treated animals (c) was considerably reduced in tumors from hrIL-27 treated animals (d). The histologic feature of poorly differentiated tumor with a solid growth pattern (e) and endowed with a well-developed vascularization (g) observed in PBS treated animals, was heavily compromised in tumors from hrIL-27 treated animals which showed multiple foci of ischemic necrosis (f) along with a deficient vascular supply (h). PC3 tumors revealed a strong expression of the anti-angiogenic chemokine IP-10/CXCL10 (j), which, on the contrary, was barely detected in tumors from PBS treated animals (i). (a-j: x400). Panel E. Expression profile of genes involved in angiogenic pathways in PC3 (white bars) and DU145 (black bars) cells treated with hrIL-27. Pooled results ± SD from two experiments performed in duplicate are shown. Histogram represents fold differences of individual mRNA between cells cultured in presence or absence of hrIL-27.

Article Snippet: Cell culture, antibodies, reagents, flow cytometry and western blot The human PC3, DU145, 22Rv1 and LNCaP PCa cell lines (LGC Standards, Teddington, UK) were cultured in RPMI 1640 with 10% FCS (Seromed-BiochromKG, Berlin, D). hrIL-27 (R&D System, Minneapolis, MN, USA) was used at 100 ng/ml following titration experiments.

Techniques: Inhibition, In Vivo, Injection, In Situ, Immunohistochemical staining, Expressing, Control, Activity Assay, Cell Culture