The preventive effect of suramin on the development of monocrotaline-induced pulmonary hypertension.

<div><p>Compared with vehicle administered alone, suramin treatment significantly prevented the development of pulmonary hypertension and right ventricular hypertrophy. <b>A</b>) Pulmonary arterial pressure measurements. <b>B</b>) Right ventricular hypertrophy as assessed by the RV/(LV+S) weight ratio. </p> <p><b>C</b>) Percentages of nonmuscular (NM), partially muscular (PM), fully muscular (FM), and completely obliterated (FM+) intra-acinar vessels. All values are the mean±SEM from at least 5 animals per group. </p> <p><b>D</b>-<b>O</b>) Histochemical and immunohistochemical analysis of rat lung tissue at 21 days after monocrotaline injection. Medial hypertrophy was associated with an increased number of proliferating vascular cells shown by immunohistochemistry for PCNA; PCNA-positive cells have dark nuclei. Suramin prevented the development of medial hypertrophy and the deposition of collagen around the pulmonary arteries. <b>D</b>-<b>F</b>) Hematoxylin-phloxine-saffron staining. <b>G</b>-<b>I</b>) Proliferation of pulmonary artery smooth muscle cells<b>. J</b>-<b>L</b>) Collagen deposition. <b>M</b>-<b>O</b>) Macrophages in the lung tissue. All values are the mean±SEM from at least 5 animals in each group. *P<0.05; **P<0.001; ***P<0.0001. The statistical analyses compared MCT-injected rats (no suramin treatment) vs. rats injected with saline instead of MCT or rats treated with suramin. Scale bar=25 µm in all sections.</p></div

Differential expression of phosphorylated growth factor receptor tyrosine kinases (RTKs) in PA-SMCs.

<p>Arrays were incubated with 150 µg of lysate prepared from untreated cells or cells treated with suramin (1000 µg/mL, 1 h) and incubated for 15 minutes with 10 ng/mL of PDGF, FGF2, or EGF. RTKs were analyzed with anti-phospho-RTK, while immunoglobulin (IgG) and "reference spots" were used as the negative and positive controls, respectively. n=4.</p

BrdU incorporation and changes in cell number in human pulmonary-artery smooth muscle cells in response to 10% fetal calf serum in the presence or absence of a range of concentrations of suramin (100 to 1000 µg/mL).

<p>n=4; *P<0.05; **P<0.001; ***P<0.0001.</p

The effect of suramin on growth factor-induced pulmonary-artery smooth muscle cell proliferation.

<p>BrdU incorporation and cell count data are shown for quiescent cells incubated with PDGF, FGF2, or EGF (10 ng/mL). Suramin (1000 µg/mL) was added 1 hour before growth factor addition. n=4; *P<0.05; **P<0.001; ***P<0.0001.</p

The structure and wall thickness of human pulmonary artery segments after 10 days of incubation in culture medium.

<p>The culture medium was either unsupplemented or supplemented with 10% FCS, suramin (1000 µg/mL) or masitinib (10^-5 M). (<b>A</b>) Hematoxylin-phloxine-saffron stain Scale bars: 25 µm (<b>B</b>) Masson trichrome stain Scale bars: 25 µm. (<b>C</b>) Double immunofluorescence staining performed with an anti-proliferating cell nuclear antigen (PCNA) antibody (red signal) and an anti-α-smooth muscle actin (α-SMA) antibody (green signal). DAPI nuclear staining is also shown (blue signal). Scale bars: 50 µm.</p

The effect of suramin on PDGF, FGF2, and EGF-induced ERK1/2 phosphorylation in PA-SMCs.

<p>Western blotting was used to assess ERK1/2 phosphorylation in PA-SMCs incubated with PDGF, FGF2, or EGF (10 ng/mL, 10 min) with or without suramin pretreatment (1000 µg/mL, 1 h). The results shown are typical findings from five independent experiments. The level of phosphorylated ERK1/2 was normalized against β-actin. n=4. *P<0.05 versus control; **P<0.001 versus control. The phospho/total ratio was expressed as the mean±SEM from five PA-SMC samples. </p

Additional file 1: Figure S1. of Osteopontin and thrombospondin-1 play opposite roles in promoting tumor aggressiveness of primary resected non-small cell lung cancer

(A). Graphical representation of the ELISA data for OPN serum levels in healthy donors (n = 20) and population study (n = 171). Figure S1 (B). Graphical representation of the ELISA data for TSP-1 serum levels in healthy donors (n = 20) and population study (n = 171). (DOCX 154 kb