Figure 3.

Mito-HNK Inhibits Invasion and Brain Metastasis of Lung Cancer Cells

(A and C) The anti-invasive effects of Mito-HNK on NSCLC lines H2030-BrM3 and PC9-BrM3 were assessed via the Boyden chamber invasion assay after 48-hr treatment. Representative images are shown in (A) and the quantitative data are shown in (C, *P < 0.05, **P < 0.01, ***P < 0.001 vs.control).

(B and D) Representative images (B) and quantitative data (D) of the invasion assay for SCLC DMS-273 cells indicate that Mito-HNK is at least 100-fold more potent than HNK (*P < 0.05, **P < 0.01 vs.control).

(E) High-resolution echocardiography to visualize the needle position within the left cardiac ventricle, for injecting lung cancer cells in the in vivo experiments to establish brain metastases.

(F) Representative bioluminescence, GFP expression, and H&E staining images of brains taken from control, HNK, and Mito-HNK (3.75 μmol/kg each) mice.

(G and H) Quantitative data for the bioluminescence imaging of brain metastases over time. Mito-HNK treatment was started 1 day after injection of H2030-BrM3 cells (G, n = 6 for control, and n = 5 for Mito-HNK, observation duration = 27 days) or DMS-273 cells (H, n = 7 each, observation duration = 14 days). (G, inset) shows the LC-MS trace from a brain extract indicating the presence of Mito-HNK (65 pmol/g of whole brain tissue). Error bars indicate SE.