BLI of untreated and treated tumor-bearing mice at different time points.

<p>Growth of lung tumors in untreated mice was characterized by initial tumor appearance in the neck region and subsequently continuous dissemination into the lungs (A). This growth pattern differed in mice which were treated with BC-819 where tumors either decreased in size or in completely vanished during treatment.</p

In-situ-hybridization for H19-activity in lung tumors.

<p>An intrabronchially induced lung tumor was stained for H.E. (A) and screened for H19-activity using ISH (B). In the given sample a strong expression of H19 was found, indicated by a brownish colorization of the examined tissue.</p

Histology of orthotopically induced lung tumors.

<p>Tumors were detectable as early as 10 days after intrabronchial implantation in the peribronchiolar region of the lungs (A). End stages of intrabronchially induced lung tumors were characterized by multiple, peribronchiolar and parenchymal localized tumors (B) and by tumor tissue within the luminal lung regions as well (C). Intravenous injection of A549-C8-luc, however, resulted in tumors predominately growing in the parenchyma of the lungs but not peribronchiolar or within the bronchi (D). Figure D shows parenchymal tumor tissue close to a bronchus, but not peribronchiolar. Alveolar tissue localized between the tumor and the bronchus however is condensed by the adjacent tumor nodule.</p

Survival of untreated and treated tumor-bearing mice.

<p>Mice bearing intrabronchially induced lung tumors showed a significant survival benefit (p = 0.01) when treated with BC-819 compared to animals which received no treatment (A). No survival benefit was observed in mice suffering from intravenously induced lung tumors, independent of treatment with BC-819 (B).</p

Effect of BC-819 on cell growth in different lung cancer cell lines.

<p>NCI-H460, NCI-H358 and A549 cells were transfected, using Lipofectamine 2000, with BC-819 and co-transfected with a plasmid, encoding for the reporter enzyme luciferase. As early as 24 h after transfection, luciferase activity (indirectly indicating cell growth) was reduced by at least >90% when 350 ng BC-819 was used (A). 48 hours later luciferase activity was decreased by more than 98%, except for the cell line NCI-H358. However, a luciferase decrease of more than 98% was observed in all cell lines when the amount of BC-819 was increased to 700 ng (B). The influence of BC-819 on bioluminescent A549-C8-luc was examined as well and revealed decreased luciferase and therefore reduced cell growth (more than 50%) as early as after 24 hours (C). Maximum inhibition of cell growth was reached by 48 hours after transfection, becoming apparent through more than 75% reduction in luciferase activity.</p