Bio-distribution of adenovirus after a subconjunctival or intravenous injection in the rats.

<p>(A) Bioluminescence images of rat at day 1, 2, 7, 14, 28, 35 and 64 after a subconjunctival or intravenous injection of Ad-Luci (1x10¹⁰ GC/eye). (B) Luciferase activity was measured in unit of photons by bioluminescence analysis. Data are presented as the means ± SEM (n = 5).</p

The serum level of IL-1β and CINC-1 in the rats received a single injection of adenoviral vector via different administration routes.

<p>Serum level of interleukin-1β (IL-1β; A) and cytokine-induced neutrophil chemoattractant factor-1 (CINC-1; B) were measured at 2 hours and 7 days after an intravenous or subconjunctival injection of Ad-Luci (1x10¹⁰ GC/eye). Data are presented as the means ± SEM (*: p<0.05, ***: p<0.0001, n = 6). One-way ANOVA followed by a Tukey’s test.</p

Analysis of retinal function by electroretinography (ERG).

<p>Analysis of retinal function by electroretinography (ERG).</p

Local ocular distribution of adenovirus after a subconjunctival injection in the rats.

<p>Histological sections of the rat eyes injected with Ad-LacZ (1x10¹⁰ GC/eye) in the subconjunctiva. The blue staining (arrows) in the image indicate the expression of β-galactosidase. V: vitreous; S: sclera. Scale bar = 200 μm (left) and 50 μm (right).</p

Effect of liver function in the rats received a single injection of adenoviral vector via different administration routes.

<p>(A) Serum level of glutamic oxaloacetic transaminase (GOT) and glutamic pyruvic transaminase (GPT) were measured at 14 days after an intravenous or subconjunctival injection of Ad-Luci (1x10¹⁰ GC/eye). Data are presented as the means ± SEM (***: p<0.0001, n = 6). One-way ANOVA followed by a Tukey’s test. (B) Sections of liver tissue were stained for TUNEL (green) and DAPI (Blue) 14 days after injection of adenovirus through different administration route. Scale bar = 200 μm.</p

Evaluation of ocular distribution and local inflammatory response after a subconjunctival injection of adenoviral vector in the rats.

<p>Evaluation of ocular distribution and local inflammatory response after a subconjunctival injection of adenoviral vector in the rats.</p

Viral dosage and duration of adenovirus-mediated gene delivery after a subconjunctival injection in the rats.

<p>(A) Bioluminescence images of rat eyes at day 1, 2, 7, 14, 28, 35 and 64 after a subconjunctival injection of Ad-Luci (1x10⁹ and 1x10¹⁰ GC/eye). (B) Luciferase activity was measured in unit of photons by bioluminescence analysis. Data are presented as the means ± SEM (n = 5).</p

Up-Regulation of Hepatoma-Derived Growth Factor Facilities Tumor Progression in Malignant Melanoma

<div><p>Cutaneous malignant melanoma is the fastest increasing malignancy in humans. Hepatoma-derived growth factor (HDGF) is a novel growth factor identified from human hepatoma cell line. HDGF overexpression is correlated with poor prognosis in various types of cancer including melanoma. However, the underlying mechanism of HDGF overexpression in developing melanoma remains unclear. In this study, human melanoma cell lines (A375, A2058, MEL-RM and MM200) showed higher levels of HDGF gene expression, whereas human epidermal melanocytes (HEMn) expressed less. Exogenous application of HDGF stimulated colony formation and invasion of human melanoma cells. Moreover, HDGF overexpression stimulated the degree of invasion and colony formation of B16–F10 melanoma cells whereas HDGF knockdown exerted opposite effects <i>in vitro</i>. To evaluate the effects of HDGF on tumour growth and metastasis <i>in vivo,</i> syngeneic mouse melanoma and metastatic melanoma models were performed by manipulating the gene expression of HDGF in melanoma cells. It was found that mice injected with HDGF-overexpressing melanoma cells had greater tumour growth and higher metastatic capability. In contrast, mice implanted with HDGF-depleted melanoma cells exhibited reduced tumor burden and lung metastasis. Histological analysis of excised tumors revealed higher degree of cell proliferation and neovascularization in HDGF-overexpressing melanoma. The present study provides evidence that HDGF promotes tumor progression of melanoma and targeting HDGF may constitute a novel strategy for the treatment of melanoma.</p> </div

Expression level of HDGF and effects of HDGF on tumorgenicity of human melanoma cells <i>in vitro.</i>

<p>(A) The gene expression of HDGF in human epidermal melanocyte (HEMn) and melanoma cell lines (A375, A2058, MEL-RM and MM200) was measured by qRT-PCR. The relative gene expression level of HDGF was normalised to GAPDH. No statistical analysis was performed (melanocyte group is populated by only one type of cell line). (B) A375 and A2058 cells were infected with adenoviral vector at different MOI (100 or 200) then evaluated gene expression level of HDGF. (C) Representative images illustrating the effects of rHDGF (10 ng/mL) and Ad-HDGF shRNA (at 200 MOI) and Ad-GFP (at 200 MOI) on colony formation identified by crystal violet stains in A375 and A2058 cells. Quantitative measures of colony formation by counting the number of crystal violet positive cells. (D) Cells invaded through polycarbonate membrane (10 mm pore size) were stained with Giemsa. Representative photomicrographs of migrated cells through the Matrigel-coated filter were quantified in A375 and A2058 cells. All data are expressed as mean ± SEM from 3 experiments. *<i>P</i><0.05 compared to control, one-way ANOVA with post-doc analysis.</p

Effects of HDGF overexpression or gene silencing on metastatic capability of melanoma cells <i>in vitro</i> and <i>in vivo.</i>

<p>(A) After infection with adenovirus vectors (Ad-GFP, Ad-HDGF and Ad-HDGF shRNA), cells were subjected to cell invasion assay. Representative photomicrographs and the degree of migrated cells through Matrigel-coated filters. (B) Live images and the quantification of photon units for 14 days were shown (n = 10/each group). All data are expressed as mean ± SEM. *<i>P</i><0.05 compared to Ad-GFP-treated groups, one-way ANOVA with post-doc analysis.</p