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Application of GFP imaging in cancer.

Hoffman RM - Lab. Invest. (2015)

Bottom Line: Non-invasive imaging with fluorescent proteins enabled the dynamics of metastatic cancer to be followed in real time in individual animals.Recent applications of the technology described here include linking fluorescent proteins with cell-cycle-specific proteins such that the cells change color from red to green as they transit from G1 to S phases.With the macro- and micro-imaging technologies described here, essentially any in vivo process can be imaged, giving rise to the new field of in vivo cell biology using fluorescent proteins.

View Article: PubMed Central - PubMed

Affiliation: AntiCancer, Inc., Department of Surgery, University of California San Diego, San Diego, CA, USA.

ABSTRACT
Multicolored proteins have allowed the color-coding of cancer cells growing in vivo and enabled the distinction of host from tumor with single-cell resolution. Non-invasive imaging with fluorescent proteins enabled the dynamics of metastatic cancer to be followed in real time in individual animals. Non-invasive imaging of cancer cells expressing fluorescent proteins has allowed the real-time determination of efficacy of candidate antitumor and antimetastatic agents in mouse models. The use of fluorescent proteins to differentially label cancer cells in the nucleus and cytoplasm can visualize the nuclear-cytoplasmic dynamics of cancer cells in vivo including: mitosis, apoptosis, cell-cycle position, and differential behavior of nucleus and cytoplasm that occurs during cancer-cell deformation and extravasation. Recent applications of the technology described here include linking fluorescent proteins with cell-cycle-specific proteins such that the cells change color from red to green as they transit from G1 to S phases. With the macro- and micro-imaging technologies described here, essentially any in vivo process can be imaged, giving rise to the new field of in vivo cell biology using fluorescent proteins.

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Interactions (arrows) of host stromal GFP-expressing fibroblast cell (arrowhead) and Dunning RFP-expressing rodent prostate cancer cells in live tumor tissue. Scale bar, 20 μm.116
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Figure 9: Interactions (arrows) of host stromal GFP-expressing fibroblast cell (arrowhead) and Dunning RFP-expressing rodent prostate cancer cells in live tumor tissue. Scale bar, 20 μm.116

Mentions: The tumor microenvironment (TME) has an important influence on tumor progression. Six different implantation models were developed to image the TME using multiple colors of fluorescent proteins: I) RFP- or GFP-expressing HCT-116 human colon cancer cells were implanted subcutaneously in the CFP-expressing nude mice. CFP stromal cells from the subcutaneous TME were visualized interacting with the RFP- or GFP-expressing tumors. II) RFP-expressing HCT-116 cells were transplanted into the spleen of CFP nude mice, which resulted in experimental metastases formed in the liver. CFP stromal cells from the liver TME were visualized interacting with the RFP-expressing tumor. III) RFP-expressing HCT-116 cancer cells were injected in the tail vein of CFP-expressing nude mice, forming experimental metastases in the lung. CFP stromal cells from the lung were visualized interacting with the RFPexpressing tumor. IV) In order to visualize two different tumors in the TME, GFP-expressing and RFP-expressing HCT-116 cancer cells were co-implanted subcutaneously in CFP-expressing nude mice. A 3-color TME was formed subcutaneously in the CFP mouse, and CFP stromal cells were visualized interacting with the RFP and GFP-expressing tumors. V) In order to have two different colors of stromal cells, GFP-expressing HCT-116 cells were initially injected subcutaneously in RFP-expressing nude mice. After 14 days, the tumor, which consisted of GFP cancer cells and RFP stromal cells derived from the RFP nude mouse, was harvested and transplanted into the CFP nude mouse. CFP stromal cells invaded the growing transplanted tumor containing GFP cancer cells and RFP stroma. VI) MMT cells expressing GFP in the nucleus and RFP in the cytoplasm were implanted in the spleen of a CFP nude mouse. Cancer cells were imaged in the liver 3 days after cell injection. The dual-color dividing MMT cells and CFP hepatocytes, as well as CFP non-parenchymal cells of the liver were imaged interacting with the 2-color cancer cells. CFP-expressing host cancer-associated fibroblasts (CAFs) (Figure 9) were predominantly observed in the TME models developed in the CFP nude mouse (Figure 10).85


Application of GFP imaging in cancer.

Hoffman RM - Lab. Invest. (2015)

Interactions (arrows) of host stromal GFP-expressing fibroblast cell (arrowhead) and Dunning RFP-expressing rodent prostate cancer cells in live tumor tissue. Scale bar, 20 μm.116
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4383682&req=5

Figure 9: Interactions (arrows) of host stromal GFP-expressing fibroblast cell (arrowhead) and Dunning RFP-expressing rodent prostate cancer cells in live tumor tissue. Scale bar, 20 μm.116
Mentions: The tumor microenvironment (TME) has an important influence on tumor progression. Six different implantation models were developed to image the TME using multiple colors of fluorescent proteins: I) RFP- or GFP-expressing HCT-116 human colon cancer cells were implanted subcutaneously in the CFP-expressing nude mice. CFP stromal cells from the subcutaneous TME were visualized interacting with the RFP- or GFP-expressing tumors. II) RFP-expressing HCT-116 cells were transplanted into the spleen of CFP nude mice, which resulted in experimental metastases formed in the liver. CFP stromal cells from the liver TME were visualized interacting with the RFP-expressing tumor. III) RFP-expressing HCT-116 cancer cells were injected in the tail vein of CFP-expressing nude mice, forming experimental metastases in the lung. CFP stromal cells from the lung were visualized interacting with the RFPexpressing tumor. IV) In order to visualize two different tumors in the TME, GFP-expressing and RFP-expressing HCT-116 cancer cells were co-implanted subcutaneously in CFP-expressing nude mice. A 3-color TME was formed subcutaneously in the CFP mouse, and CFP stromal cells were visualized interacting with the RFP and GFP-expressing tumors. V) In order to have two different colors of stromal cells, GFP-expressing HCT-116 cells were initially injected subcutaneously in RFP-expressing nude mice. After 14 days, the tumor, which consisted of GFP cancer cells and RFP stromal cells derived from the RFP nude mouse, was harvested and transplanted into the CFP nude mouse. CFP stromal cells invaded the growing transplanted tumor containing GFP cancer cells and RFP stroma. VI) MMT cells expressing GFP in the nucleus and RFP in the cytoplasm were implanted in the spleen of a CFP nude mouse. Cancer cells were imaged in the liver 3 days after cell injection. The dual-color dividing MMT cells and CFP hepatocytes, as well as CFP non-parenchymal cells of the liver were imaged interacting with the 2-color cancer cells. CFP-expressing host cancer-associated fibroblasts (CAFs) (Figure 9) were predominantly observed in the TME models developed in the CFP nude mouse (Figure 10).85

Bottom Line: Non-invasive imaging with fluorescent proteins enabled the dynamics of metastatic cancer to be followed in real time in individual animals.Recent applications of the technology described here include linking fluorescent proteins with cell-cycle-specific proteins such that the cells change color from red to green as they transit from G1 to S phases.With the macro- and micro-imaging technologies described here, essentially any in vivo process can be imaged, giving rise to the new field of in vivo cell biology using fluorescent proteins.

View Article: PubMed Central - PubMed

Affiliation: AntiCancer, Inc., Department of Surgery, University of California San Diego, San Diego, CA, USA.

ABSTRACT
Multicolored proteins have allowed the color-coding of cancer cells growing in vivo and enabled the distinction of host from tumor with single-cell resolution. Non-invasive imaging with fluorescent proteins enabled the dynamics of metastatic cancer to be followed in real time in individual animals. Non-invasive imaging of cancer cells expressing fluorescent proteins has allowed the real-time determination of efficacy of candidate antitumor and antimetastatic agents in mouse models. The use of fluorescent proteins to differentially label cancer cells in the nucleus and cytoplasm can visualize the nuclear-cytoplasmic dynamics of cancer cells in vivo including: mitosis, apoptosis, cell-cycle position, and differential behavior of nucleus and cytoplasm that occurs during cancer-cell deformation and extravasation. Recent applications of the technology described here include linking fluorescent proteins with cell-cycle-specific proteins such that the cells change color from red to green as they transit from G1 to S phases. With the macro- and micro-imaging technologies described here, essentially any in vivo process can be imaged, giving rise to the new field of in vivo cell biology using fluorescent proteins.

Show MeSH
Related in: MedlinePlus