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Automated whole animal bio-imaging assay for human cancer dissemination.

Ghotra VP, He S, de Bont H, van der Ent W, Spaink HP, van de Water B, Snaar-Jagalska BE, Danen EH - PLoS ONE (2012)

Bottom Line: Findings in this model correlate with behavior in long-term rodent xenograft models for panels of poorly- versus highly malignant cell lines derived from breast, colorectal, and prostate cancer.Moreover, RNA interference establishes the metastasis-suppressor role for E-cadherin in this model.This automated quantitative whole animal bio-imaging assay can serve as a first-line in vivo screening step in the anti-cancer drug target discovery pipeline.

View Article: PubMed Central - PubMed

Affiliation: Division of Toxicology, Leiden/Amsterdam Center for Drug Research, Leiden University, Leiden, the Netherlands.

ABSTRACT
A quantitative bio-imaging platform is developed for analysis of human cancer dissemination in a short-term vertebrate xenotransplantation assay. Six days after implantation of cancer cells in zebrafish embryos, automated imaging in 96 well plates coupled to image analysis algorithms quantifies spreading throughout the host. Findings in this model correlate with behavior in long-term rodent xenograft models for panels of poorly- versus highly malignant cell lines derived from breast, colorectal, and prostate cancer. In addition, cancer cells with scattered mesenchymal characteristics show higher dissemination capacity than cell types with epithelial appearance. Moreover, RNA interference establishes the metastasis-suppressor role for E-cadherin in this model. This automated quantitative whole animal bio-imaging assay can serve as a first-line in vivo screening step in the anti-cancer drug target discovery pipeline.

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Related in: MedlinePlus

Determination of cancer cell dissemination kinetics.(A and B) LnCAP (A) or PC3 cells (B) were implanted and embryos were fixed at 2, 4, or 6 dpi for imaging (immunofluorescence images and automated image analysis (scatter plots)). Bottom row images (scale bar = 50 µm) show zoom-in of area marked by dotted line in top row images (scale bar = 100 µm). (C) CD at 2, 4 and 6 dpi for LnCAP (grey) and PC3-injected embryos (black) calculated from scatterplots in A and B, respectively. Statistical testing for difference between LnCAP and PC3 at different dpi is indicated. *p<0.05, ***p<0.001.
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pone-0031281-g004: Determination of cancer cell dissemination kinetics.(A and B) LnCAP (A) or PC3 cells (B) were implanted and embryos were fixed at 2, 4, or 6 dpi for imaging (immunofluorescence images and automated image analysis (scatter plots)). Bottom row images (scale bar = 50 µm) show zoom-in of area marked by dotted line in top row images (scale bar = 100 µm). (C) CD at 2, 4 and 6 dpi for LnCAP (grey) and PC3-injected embryos (black) calculated from scatterplots in A and B, respectively. Statistical testing for difference between LnCAP and PC3 at different dpi is indicated. *p<0.05, ***p<0.001.

Mentions: We performed experiments to determine the earliest time point that allowed robust discrimination between poorly aggressive and highly aggressive cell lines. Using LnCAP and PC3 as such an example for prostate cancer [14]–[19] we observed no difference at 2 dpi; MCD of PC3 could be distinguished from MCD of LnCAP at 4 dpi; and at 6 dpi MCD was markedly higher in PC3 compared to LnCAP with strong significance (Figure 4). Therefore, 6 dpi was chosen for analysis in all further experiments.


Automated whole animal bio-imaging assay for human cancer dissemination.

Ghotra VP, He S, de Bont H, van der Ent W, Spaink HP, van de Water B, Snaar-Jagalska BE, Danen EH - PLoS ONE (2012)

Determination of cancer cell dissemination kinetics.(A and B) LnCAP (A) or PC3 cells (B) were implanted and embryos were fixed at 2, 4, or 6 dpi for imaging (immunofluorescence images and automated image analysis (scatter plots)). Bottom row images (scale bar = 50 µm) show zoom-in of area marked by dotted line in top row images (scale bar = 100 µm). (C) CD at 2, 4 and 6 dpi for LnCAP (grey) and PC3-injected embryos (black) calculated from scatterplots in A and B, respectively. Statistical testing for difference between LnCAP and PC3 at different dpi is indicated. *p<0.05, ***p<0.001.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC3275564&req=5

pone-0031281-g004: Determination of cancer cell dissemination kinetics.(A and B) LnCAP (A) or PC3 cells (B) were implanted and embryos were fixed at 2, 4, or 6 dpi for imaging (immunofluorescence images and automated image analysis (scatter plots)). Bottom row images (scale bar = 50 µm) show zoom-in of area marked by dotted line in top row images (scale bar = 100 µm). (C) CD at 2, 4 and 6 dpi for LnCAP (grey) and PC3-injected embryos (black) calculated from scatterplots in A and B, respectively. Statistical testing for difference between LnCAP and PC3 at different dpi is indicated. *p<0.05, ***p<0.001.
Mentions: We performed experiments to determine the earliest time point that allowed robust discrimination between poorly aggressive and highly aggressive cell lines. Using LnCAP and PC3 as such an example for prostate cancer [14]–[19] we observed no difference at 2 dpi; MCD of PC3 could be distinguished from MCD of LnCAP at 4 dpi; and at 6 dpi MCD was markedly higher in PC3 compared to LnCAP with strong significance (Figure 4). Therefore, 6 dpi was chosen for analysis in all further experiments.

Bottom Line: Findings in this model correlate with behavior in long-term rodent xenograft models for panels of poorly- versus highly malignant cell lines derived from breast, colorectal, and prostate cancer.Moreover, RNA interference establishes the metastasis-suppressor role for E-cadherin in this model.This automated quantitative whole animal bio-imaging assay can serve as a first-line in vivo screening step in the anti-cancer drug target discovery pipeline.

View Article: PubMed Central - PubMed

Affiliation: Division of Toxicology, Leiden/Amsterdam Center for Drug Research, Leiden University, Leiden, the Netherlands.

ABSTRACT
A quantitative bio-imaging platform is developed for analysis of human cancer dissemination in a short-term vertebrate xenotransplantation assay. Six days after implantation of cancer cells in zebrafish embryos, automated imaging in 96 well plates coupled to image analysis algorithms quantifies spreading throughout the host. Findings in this model correlate with behavior in long-term rodent xenograft models for panels of poorly- versus highly malignant cell lines derived from breast, colorectal, and prostate cancer. In addition, cancer cells with scattered mesenchymal characteristics show higher dissemination capacity than cell types with epithelial appearance. Moreover, RNA interference establishes the metastasis-suppressor role for E-cadherin in this model. This automated quantitative whole animal bio-imaging assay can serve as a first-line in vivo screening step in the anti-cancer drug target discovery pipeline.

Show MeSH
Related in: MedlinePlus