Comparison of B16-luc, LLC1-luc and TRAMP-C2-luc tumor cell dissemination versus microsphere dissemination (7 and 16 μm diameter) in osseous organs.

<p>(A) Only ~5% of tumor cells and microbeads disseminated to the spine. No significant difference was found between the groups. (B) Long bones revealed dissemination fractions between ~10–25%. TRAMP-C2-luc disseminated significantly higher to long bones compared to LLC1-luc cells (LLC1-luc vs. TRAMP-C2-luc: p = 0.0406). No other significant dissemination differences were found. (C) Low tumor cell and microbeads dissemination was found in the thorax (~5–10%). No significant difference was found between the groups. (D) Dissemination to the cranium was ~2–6%. No significant dissemination differences were found. Mean ± SD, (B16-luc n = 4, LLC1-luc n = 5, TRAMP-C2-luc n = 5, 7 μm beads n = 5, 16 μm beads n = 5) for all experiments shown, one way ANOVA with Bonferroni post hoc analysis.</p

B16-luc, LLC1-luc, TRAMP-C2-luc and microsphere dissemination (7 and 16 μm diameter) in soft tissues.

<p>(A) Individual lung dissemination of tumor cells varied between ~30–40%. In contrast, very few microbeads (independent of their size) were found in the lung. All cell types disseminated significantly higher to the lung compared to microbeads (B16-luc vs. 7 μm beads: p < 0.0001, B16-luc vs. 16 μm beads: p < 0.0001, LLC1-luc vs. 7 μm beads: p = 0.0025, LLC1-luc vs. 16 μm beads: p = 0.0039, TRAMP-C2-luc vs. 7 μm beads: p = 0.0024, TRAMP-C2-luc vs. 16 μm beads: p = 0.0037). (B) Tumor cell and microbeads dissemination to the brain was relatively low (~5%). LLC1-luc cells disseminated significantly higher to the brain compared to B16-luc. Cell sized (16 μm) microbeads also disseminated significantly higher to the brain compared to B16-luc and even TRAMP-C2-luc (B16-luc vs. LLC1-luc: p = 0.0288, B16-luc vs. 7 μm beads: p = 0.0386, B16-luc vs. 16 μm beads: p = 0.0012, TRAMP-C2-luc vs. 16 μm beads: p = 0.0019). (C) The kidneys’ dissemination fraction ranged from ~10 to 25%. Small (7 μm) microbeads disseminated significantly higher to the kidneys compared to B16-luc and LLC1-luc tumor cells (LLC1-luc vs. 7μm beads: p = 0.0327). (D) Microbeads dissemination was high in the liver (~20%), whereas cell dissemination varied by cell type. LLC1-luc cells disseminated significantly stronger to the liver compared to the other cells. Comparably high amounts of microbeads were found in the liver, 7 μm beads disseminated significantly higher to the liver compared to B16-luc and TRAMP-C2, 16 μm beads showed significantly higher dissemination compared to TRAMP-C2 (B16-luc vs. LLC1-luc: p = 0.0029, B16-luc vs. 7μm beads: p = 0.0116, B16-luc vs. 16μm beads: p = 0.0338, LLC1-luc vs. TRAMP-C2-luc: p < 0.0001, TRAMP-C2-luc vs. 7μm beads: p = 0.0003, TRAMP-C2-luc vs. 16μm beads: p = 0.0003). (E) The skin showed high dissemination of microbeads and TRAMP-C2-luc cells, with a significant effect compared to B16-luc cells (B16-luc vs. TRAMP-C2-luc: p = 0.0366, B16-luc vs. 7μm beads: p = 0.0082, B16-luc vs. 16μm beads: p = 0.0084). Mean ± SD, (B16-luc n = 4, LLC1-luc n = 5, TRAMP-C2-luc n = 5, 7 μm beads n = 5, 16 μm beads n = 5) for all experiments shown, one way ANOVA with Bonferroni post hoc analysis.</p

Characterization of firefly luciferase—eGFP infected B16-F1 melanoma, LLC1 lung carcinoma and TRAMP-C2 prostate carcinoma cells.

<p>(A) Cells were lentivirally transduced to express firefly luciferase, enhanced GFP and puromycin. Representative phase contrast and green fluorescence image of B16-luc, LLC1-luc and TRAMP-C2-luc cells (bar represents 100 μm). (B) Analysis of luciferase activity by spectrometer demonstrates relative light units (RLU) emission by lysed cells. Bars represent mean ± SD, Student’s two tailed t-test (B16-luc vs. B16: p = < 0.0001, n = 7, LLC1-luc vs. LLC1: p = < 0.0001, n = 6, TRAMP-C2-luc vs. TRAMP-C2: p = < 0.0001, n = 6). (C) Viability was measured by absorbance of MTT. The absorbance was significantly higher in B16-luc cells seeded at 10’000 cells per well compared to control B16 cells (p < 0.0001). There was no viability difference in LLC1-luc and TRAMP-C2-luc cells compared to LLC1 and TRMAP-C2 cells respectively. Mean ± SD, n = 5 for all experiments shown, two way ANOVA with Bonferroni post hoc analysis. (D) Wound healing 2D migration analysis shows no difference in B16-luc, LLC1-luc andTRAMP-C2-luc cell migration 24 hours post scratch. Mean ± SD, n = 3 for all experiments shown, two way ANOVA with Bonferroni post hoc analysis.</p