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In vivo lymphatic imaging of a human inflammatory breast cancer model.

Agollah GD, Wu G, Sevick-Muraca EM, Kwon S - J Cancer (2014)

Bottom Line: Inflammatory breast cancer (IBC) remains the most aggressive type of breast cancer with the greatest potential for metastasis and as a result, the highest mortality rate.Herein, we non-invasively and longitudinally imaged lymphatics in an animal model of IBC using near-infrared fluorescence (NIRF) imaging.We also observed increased and dilated fluorescent lymphatic vessels in the tumor periphery, which was confirmed by ex vivo immunohistochemical staining of lymphatic vessels.

View Article: PubMed Central - PubMed

Affiliation: 1. Center for Molecular Imaging, The Brown Foundation Institute of Molecular Medicine, The University of Texas Health Science Center, Houston, TX 77030; ; 2. The University of Texas Graduate School of Biomedical Sciences at Houston. The University of Texas MD Anderson Cancer Center, Houston, Texas 77030.

ABSTRACT

Background: Inflammatory breast cancer (IBC) remains the most aggressive type of breast cancer with the greatest potential for metastasis and as a result, the highest mortality rate. IBC cells invade and metastasize through dermal lymphatic vessels; however, it is unknown how lymphatic drainage patterns change during IBC growth and metastasis. Herein, we non-invasively and longitudinally imaged lymphatics in an animal model of IBC using near-infrared fluorescence (NIRF) imaging.

Materials and methods: Mice were imaged in vivo prior to, and up to 11 weeks after subcutaneous or orthotopic inoculation of human IBC SUM149 cells, which were stably transfected with infrared fluorescence protein (iRFP) gene reporter (SUM149-iRFP), following intradermal (i.d.) injection of indocyanine green (ICG).

Results: Fluorescence images showed well-defined lymphatic vessels prior to SUM149-iRFP inoculation. However, altered lymphatic drainage patterns including rerouting of lymphatic drainage were detected in mice with SUM149-iRFP, due to lymphatic obstruction of normal lymphatic drainages caused by tumor growth. In addition, we observed tortuous lymphatic vessels and extravasation of ICG-laden lymph in mice with SUM149-iRFP. We also observed increased and dilated fluorescent lymphatic vessels in the tumor periphery, which was confirmed by ex vivo immunohistochemical staining of lymphatic vessels.

Conclusions: Our pre-clinical studies demonstrate that non-invasive NIRF imaging can provide a method to assess changes in lymphatic drainage patterns during IBC growth and metastasis.

No MeSH data available.


Related in: MedlinePlus

(A) Bright field, iRFP fluorescence, and an overlay of SUM149 transfected with iRFP showing red cytoplasmic fluorescent signal. (B) FACS analysis showing >80% iRFP expression in SUM149. Scale, 200 µm.
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Figure 1: (A) Bright field, iRFP fluorescence, and an overlay of SUM149 transfected with iRFP showing red cytoplasmic fluorescent signal. (B) FACS analysis showing >80% iRFP expression in SUM149. Scale, 200 µm.

Mentions: SUM149 cells were transfected with piRFP plasmid to obtain fluorescent cells. After transfection, cells were selected with medium that contained 1200 μg/mL gentamicin (G418). Subclones with high iRFP-expression were isolated in 96-well plates by limiting dilution. iRFP expression was confirmed by immunofluorescence (Figure 1A) and FACs analysis (Figure 1B). There was no difference in the cell proliferation rates of parental cells and selected RFP transfectants, as determined by comparing their growth rate in monolayer culture (data not shown).


In vivo lymphatic imaging of a human inflammatory breast cancer model.

Agollah GD, Wu G, Sevick-Muraca EM, Kwon S - J Cancer (2014)

(A) Bright field, iRFP fluorescence, and an overlay of SUM149 transfected with iRFP showing red cytoplasmic fluorescent signal. (B) FACS analysis showing >80% iRFP expression in SUM149. Scale, 200 µm.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC4216802&req=5

Figure 1: (A) Bright field, iRFP fluorescence, and an overlay of SUM149 transfected with iRFP showing red cytoplasmic fluorescent signal. (B) FACS analysis showing >80% iRFP expression in SUM149. Scale, 200 µm.
Mentions: SUM149 cells were transfected with piRFP plasmid to obtain fluorescent cells. After transfection, cells were selected with medium that contained 1200 μg/mL gentamicin (G418). Subclones with high iRFP-expression were isolated in 96-well plates by limiting dilution. iRFP expression was confirmed by immunofluorescence (Figure 1A) and FACs analysis (Figure 1B). There was no difference in the cell proliferation rates of parental cells and selected RFP transfectants, as determined by comparing their growth rate in monolayer culture (data not shown).

Bottom Line: Inflammatory breast cancer (IBC) remains the most aggressive type of breast cancer with the greatest potential for metastasis and as a result, the highest mortality rate.Herein, we non-invasively and longitudinally imaged lymphatics in an animal model of IBC using near-infrared fluorescence (NIRF) imaging.We also observed increased and dilated fluorescent lymphatic vessels in the tumor periphery, which was confirmed by ex vivo immunohistochemical staining of lymphatic vessels.

View Article: PubMed Central - PubMed

Affiliation: 1. Center for Molecular Imaging, The Brown Foundation Institute of Molecular Medicine, The University of Texas Health Science Center, Houston, TX 77030; ; 2. The University of Texas Graduate School of Biomedical Sciences at Houston. The University of Texas MD Anderson Cancer Center, Houston, Texas 77030.

ABSTRACT

Background: Inflammatory breast cancer (IBC) remains the most aggressive type of breast cancer with the greatest potential for metastasis and as a result, the highest mortality rate. IBC cells invade and metastasize through dermal lymphatic vessels; however, it is unknown how lymphatic drainage patterns change during IBC growth and metastasis. Herein, we non-invasively and longitudinally imaged lymphatics in an animal model of IBC using near-infrared fluorescence (NIRF) imaging.

Materials and methods: Mice were imaged in vivo prior to, and up to 11 weeks after subcutaneous or orthotopic inoculation of human IBC SUM149 cells, which were stably transfected with infrared fluorescence protein (iRFP) gene reporter (SUM149-iRFP), following intradermal (i.d.) injection of indocyanine green (ICG).

Results: Fluorescence images showed well-defined lymphatic vessels prior to SUM149-iRFP inoculation. However, altered lymphatic drainage patterns including rerouting of lymphatic drainage were detected in mice with SUM149-iRFP, due to lymphatic obstruction of normal lymphatic drainages caused by tumor growth. In addition, we observed tortuous lymphatic vessels and extravasation of ICG-laden lymph in mice with SUM149-iRFP. We also observed increased and dilated fluorescent lymphatic vessels in the tumor periphery, which was confirmed by ex vivo immunohistochemical staining of lymphatic vessels.

Conclusions: Our pre-clinical studies demonstrate that non-invasive NIRF imaging can provide a method to assess changes in lymphatic drainage patterns during IBC growth and metastasis.

No MeSH data available.


Related in: MedlinePlus