Journal: Biology Open

Article Title: The role of macrophage migration inhibitory factor in promoting benign prostatic hyperplasia epithelial cell growth by modulating COX-2 and P53 signaling

doi: 10.1242/bio.053447

Figure Lengend Snippet: MIF promoted proliferation of BPH-1 and PWR-1E cells. (A) MTT assay showed the proliferation of BPH-1 cells treated with various concentrations of rMIF for 3 days. Data are presented as mean±s.d., n =3. (B) CCK8 assay showed the proliferation of BPH-1 cells treated with control, rMIF (100 ng/ml), rMIF (100 ng/ml)+ISO-1 (10 µM) and ISO-1 (10 µM), respectively. Data are presented as mean±s.d., n =3. (C) CCK8 assay showed the proliferation of PWR-1E cells treated with control, rMIF, rMIF+ISO-1 and ISO-1, respectively. Data are presented as mean±s.d., n =3. (D) JC-1 assay showed the growth rates of the BPH-1 cells treated by control, rMIF, rMIF+ISO-1 and ISO-1, respectively. Data are presented as mean±s.d., n =3. (E) JC-1 assay showed the growth rates of the PWR-1E cells treated by control, rMIF, rMIF+ISO-1 and ISO-1, respectively. Data are presented as mean±s.d., n =3. * P <0.05, * P <0.05. Statistical analyses were performed using ANOVA followed by Tukey's test for multiple comparison.

Article Snippet: One hundred thousand BPH-1 and PWR-1E cells were seeded into each well of a six-well plate, and rMIF (300-69, PeproTech, USA; 100 ng/ml), ISO-1 (S7732, Selleck, USA; 10 μM), celecoxib (S1261, Selleck, USA; 5 μM) or nothing was added into each well.

Techniques: MTT Assay, CCK-8 Assay, Control, Comparison

Journal: Biology Open

Article Title: The role of macrophage migration inhibitory factor in promoting benign prostatic hyperplasia epithelial cell growth by modulating COX-2 and P53 signaling

doi: 10.1242/bio.053447

Figure Lengend Snippet: Flow cytometry test that MIF promoted proliferation of BPH-1 and PWR-1E cells. (A) Flow cytometry showed cell cycle in BPH-1 cells treated by control, rMIF, rMIF+ISO-1 and ISO-1, respectively. (B) Histogram for S+G2/M phase of the cell cycle results in BPH-1 cells. Data are presented as mean±s.d., n =3. (C) Flow cytometry showed cell cycle in PWR-1E cells treated by control, rMIF, rMIF+ISO-1 and ISO-1, respectively. (D) Histogram for S+G2/M phase of the cell cycle results in PWR-1E cells. Data are presented as mean±s.d., n =3. * P <0.05. Statistical analyses were performed using ANOVA followed by Tukey's test for multiple comparison.

Article Snippet: One hundred thousand BPH-1 and PWR-1E cells were seeded into each well of a six-well plate, and rMIF (300-69, PeproTech, USA; 100 ng/ml), ISO-1 (S7732, Selleck, USA; 10 μM), celecoxib (S1261, Selleck, USA; 5 μM) or nothing was added into each well.

Techniques: Flow Cytometry, Control, Comparison

Journal: Biology Open

Article Title: The role of macrophage migration inhibitory factor in promoting benign prostatic hyperplasia epithelial cell growth by modulating COX-2 and P53 signaling

doi: 10.1242/bio.053447

Figure Lengend Snippet: MIF modulation COX-2 and P53 signaling pathway. (A) Western blotting detected the expression of COX-2 and P53 in BPH-1 cells treated by control, rMIF and rMIF+ISO-1, respectively. (B) Western blot analysis: the expression of COX-2 and P53 in BPH-1 cells treated by control, rMIF and rMIF+ISO-1. Data are presented as mean±s.d., n =3. (C) Western blotting detected the expression of COX-2 and P53 in PWR-1E cells treated by control, rMIF and rMIF+ISO-1, respectively. (D) Western blot analysis: the expression of COX-2 and P53 in PWR-1E cells treated by control, rMIF and rMIF+ISO-1. Data are presented as mean±s.d., n =3. * P <0.05. Statistical analyses were performed using ANOVA followed by Tukey's test for multiple comparison.

Article Snippet: One hundred thousand BPH-1 and PWR-1E cells were seeded into each well of a six-well plate, and rMIF (300-69, PeproTech, USA; 100 ng/ml), ISO-1 (S7732, Selleck, USA; 10 μM), celecoxib (S1261, Selleck, USA; 5 μM) or nothing was added into each well.

Techniques: Western Blot, Expressing, Control, Comparison

Journal: Biology Open

Article Title: The role of macrophage migration inhibitory factor in promoting benign prostatic hyperplasia epithelial cell growth by modulating COX-2 and P53 signaling

doi: 10.1242/bio.053447

Figure Lengend Snippet: COX-2 is a key factor of MIF promoted proliferation. (A) CCK8 assay showed the proliferation of BPH-1 cells treated with control, rMIF (100 ng/ml), rMIF (100 ng/ml)+celecoxib (5 µM) and celecoxib (5 µM), respectively. Data are presented as mean±s.d., n =3. (B) CCK8 assay showed the proliferation of PWR-1E cells treated with control, rMIF, rMIF+celecoxib and celecoxib, respectively. Data are presented as mean±s.d., n =3. (C) JC-1 assay showed the growth rates of the BPH-1 cells treated by control, rMIF, rMIF+celecoxib and celecoxib, respectively. Data are presented as mean±s.d., n =3. (D) JC-1 assay showed the growth rates of the PWR-1E cells treated by control, rMIF, rMIF+celecoxib and celecoxib, respectively. Data are presented as mean±s.d., n =3. * P <0.05. Statistical analyses were performed using ANOVA followed by Tukey's test for multiple comparison.

Article Snippet: One hundred thousand BPH-1 and PWR-1E cells were seeded into each well of a six-well plate, and rMIF (300-69, PeproTech, USA; 100 ng/ml), ISO-1 (S7732, Selleck, USA; 10 μM), celecoxib (S1261, Selleck, USA; 5 μM) or nothing was added into each well.

Techniques: CCK-8 Assay, Control, Comparison

Journal: Biology Open

Article Title: The role of macrophage migration inhibitory factor in promoting benign prostatic hyperplasia epithelial cell growth by modulating COX-2 and P53 signaling

doi: 10.1242/bio.053447

Figure Lengend Snippet: Flow cytometry test that COX-2 is a key factor of MIF promoted proliferation. (A) Flow cytometry showed cell cycle in BPH-1 cells treated by control, rMIF, rMIF+celecoxib and celecoxib, respectively. (B) Histogram for S+G2/M phase of the cell cycle results in BPH-1 cells. Data are presented as mean±s.d., n =3. (C) Flow cytometry showed cell cycle in PWR-1E cells treated by control, rMIF, rMIF+celecoxib and celecoxib, respectively. (D) Histogram for S+G2/M phase of the cell cycle results in PWR-1E cells. Data are presented as mean±s.d., n =3. * P <0.05. Statistical analyses were performed using ANOVA followed by Tukey's test for multiple comparison.

Article Snippet: One hundred thousand BPH-1 and PWR-1E cells were seeded into each well of a six-well plate, and rMIF (300-69, PeproTech, USA; 100 ng/ml), ISO-1 (S7732, Selleck, USA; 10 μM), celecoxib (S1261, Selleck, USA; 5 μM) or nothing was added into each well.

Techniques: Flow Cytometry, Control, Comparison

Journal: Biology Open

Article Title: The role of macrophage migration inhibitory factor in promoting benign prostatic hyperplasia epithelial cell growth by modulating COX-2 and P53 signaling

doi: 10.1242/bio.053447

Figure Lengend Snippet: Expression of signaling molecules COX-2 and P53 involved in the inhibition of COX-2. (A) Western blotting detected the expression of COX-2 and P53 in BPH-1 cells treated by control, rMIF and rMIF+celecoxib, respectively. (B) Western blot analysis: the expression of COX-2 and P53 in BPH-1 cells treated by control, rMIF and rMIF+celecoxib. Data are presented as mean±s.d., n =3. (C) Western blotting detected the expression of COX-2 and P53 in PWR-1E cells treated by control, rMIF and rMIF+celecoxib, respectively. (D) Western blot analysis: the expression of COX-2 and P53 in PWR-1E cells treated by control, rMIF and rMIF+celecoxib. Data are presented as mean±s.d., n =3. * P <0.05. Statistical analyses were performed using ANOVA followed by Tukey's test for multiple comparison.

Article Snippet: One hundred thousand BPH-1 and PWR-1E cells were seeded into each well of a six-well plate, and rMIF (300-69, PeproTech, USA; 100 ng/ml), ISO-1 (S7732, Selleck, USA; 10 μM), celecoxib (S1261, Selleck, USA; 5 μM) or nothing was added into each well.

Techniques: Expressing, Inhibition, Western Blot, Control, Comparison

Journal: BMC Microbiology

Article Title: Porphyromonas gingivalis ATCC 33277 promotes intercellular adhesion molecule-1 expression in endothelial cells and monocyte-endothelial cell adhesion through macrophage migration inhibitory factor

doi: 10.1186/s12866-018-1156-1

Figure Lengend Snippet: Macrophage migration-inhibitory factor (MIF) acted as a regulator in Porphyromonas gingivalis ( P. gingivalis )-induced intercellular adhesion molecule-1 (ICAM-1) expression. a Western blot analysis of ICAM-1 in EA.hy926 cells. b Quantitative analyses of the optical density relative to the internal reference protein GAPDH in Western blot analysis. c Quantitative Real-time PCR analysis of ICAM-1 mRNA. Infection with P. gingivalis ATCC 33277 (MOI = 100, 24 h) could induce the expression of ICAM-1 and ICAM-1 mRNA in EA.hy926 cells, this inductive effect of P. gingivalis was blocked by the MIF antagonist ISO-1. Sufficient exogenous recombinant MIF (rMIF) supplementation rescued the ICAM-1 expression. Data are presented as means ± SD from three independent experiments. * P < 0.01

Article Snippet: The EA.hy926 cells were pretreated with the MIF antagonist ISO-1 (25 μM; Cayman Chemical) or human rMIF (0.5 μg/mL; Cayman Chemica) plus ISO-1 for 1 h; then, the cells were infected with P. gingivalis ATCC 33277 at an MOI of 100 for 24 h. The whole cell protein of EA.hy926 cells was extracted, and Western blotting was performed.

Techniques: Migration, Expressing, Western Blot, Real-time Polymerase Chain Reaction, Infection, Recombinant

Journal: BMC Microbiology

Article Title: Porphyromonas gingivalis ATCC 33277 promotes intercellular adhesion molecule-1 expression in endothelial cells and monocyte-endothelial cell adhesion through macrophage migration inhibitory factor

doi: 10.1186/s12866-018-1156-1

Figure Lengend Snippet: Macrophage migration-inhibitory factor (MIF) regulated the increased monocyte adhesion to endothelial cells infected with Porphyromonas gingivalis ( P. gingivalis ) . a THP-1 cell adhesion to EA.hy926 cells was labeled by calcein-AM and visualized. Pictures are representative fields captured by flurencence microscope (upper line) or microscope (lower line) of three independent experiments (magnification × 100). b The histogram of the evaluation of adhered THP-1 cell assessed by cell count assay. Compared with uninfected cells, P. gingivalis ATCC 33277 infection (MOI = 100, 24 h) markedly increased THP-1 cell adhesion to endothelial cells ( P < 0.01). In contrast, cell adhesion was decreased in ISO-1-treated cells compared with those infected with P. gingivalis ATCC 33277 ( P < 0.01). And THP-1 cell adhesion to EA.hy926 cells was recovered by exogenous rMIF addition. * P < 0.01. Scale bar = 100 μm

Article Snippet: The EA.hy926 cells were pretreated with the MIF antagonist ISO-1 (25 μM; Cayman Chemical) or human rMIF (0.5 μg/mL; Cayman Chemica) plus ISO-1 for 1 h; then, the cells were infected with P. gingivalis ATCC 33277 at an MOI of 100 for 24 h. The whole cell protein of EA.hy926 cells was extracted, and Western blotting was performed.

Techniques: Migration, Infection, Labeling, Microscopy, Cell Counting