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Novel mouse mammary cell lines for in vivo bioluminescence imaging (BLI) of bone metastasis.

Bolin C, Sutherland C, Tawara K, Moselhy J, Jorcyk CL - Biol Proced Online (2012)

Bottom Line: The 4 T1.2 luc3 cell line was found to closely model the sites of metastases seen in human patients including lung, liver, and bone.High osteolytic activity of the 4 T1.2 luc3 cells in vivo in the bone microenvironment was also detected.The engineered 4 T1.2 luc3 and 66c14 luc2 cell lines described in this study are valuable tools for studying the cellular events moderating the metastasis of breast tumor cells to bone.

View Article: PubMed Central - HTML - PubMed

Affiliation: Department of Biological Sciences, Boise State University, Boise, ID, USA. cjorcyk@boisestate.edu.

ABSTRACT

Background: Tumor cell lines that can be tracked in vivo during tumorigenesis and metastasis provide vital tools for studying the specific cellular mechanisms that mediate these processes as well as investigating therapeutic targets to inhibit them. The goal of this study was to engineer imageable mouse mammary tumor cell lines with discrete propensities to metastasize to bone in vivo. Two novel luciferase expressing cell lines were developed and characterized for use in the study of breast cancer metastasis to bone in a syngeneic mouse model.

Results: The 4 T1.2 luc3 and 66c14 luc2 cell lines were shown to have high levels of bioluminescence intensity in vitro and in vivo after orthotopic injection into mouse mammary fat pads. The 4 T1.2 luc3 cell line was found to closely model the sites of metastases seen in human patients including lung, liver, and bone. Specifically, 4 T1.2 luc3 cells demonstrated a high incidence of metastasis to spine, with an ex-vivo BLI intensity three orders of magnitude above the commercially available 4 T1 luc2 cells. 66c14 luc2 cells also demonstrated metastasis to spine, which was lower than that of 4 T1.2 luc3 cells but higher than 4 T1 luc2 cells, in addition to previously unreported metastases in the liver. High osteolytic activity of the 4 T1.2 luc3 cells in vivo in the bone microenvironment was also detected.

Conclusions: The engineered 4 T1.2 luc3 and 66c14 luc2 cell lines described in this study are valuable tools for studying the cellular events moderating the metastasis of breast tumor cells to bone.

No MeSH data available.


Related in: MedlinePlus

4T1.2 luc3 cells are osteolytic in vivo. (A)  1 × 104 4 T1.2 luc3 (left) or 4 T1 luc2 cells (right) were injected into the proximal end of the left tibia of a 6-week old female Balb/c mice and visualized after i.p. injection of 150 mg/kg D-Luciferin using an IVIS Spectrum BLI system (day 18 after tumor cell injection). Whole body ventral view shows luciferase activity from tumor cells in the left tibia, validated with ex-vivo imaging of the tibia. (B) micro-CT images of cross sectional slices (4000 × 4000 pixels) (i) at the proximal and (ii) distal ends of the tibia reveal extensive osteolytic bone degradation (osteolytic damage highlighted with white arrows) near the injection site. Micro-CT images scanned using a Microphotonics Skyscan 1172 CT at a 6μm resolution and reconstructed using ImageVis3D imaging software.
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Figure 7: 4T1.2 luc3 cells are osteolytic in vivo. (A) 1 × 104 4 T1.2 luc3 (left) or 4 T1 luc2 cells (right) were injected into the proximal end of the left tibia of a 6-week old female Balb/c mice and visualized after i.p. injection of 150 mg/kg D-Luciferin using an IVIS Spectrum BLI system (day 18 after tumor cell injection). Whole body ventral view shows luciferase activity from tumor cells in the left tibia, validated with ex-vivo imaging of the tibia. (B) micro-CT images of cross sectional slices (4000 × 4000 pixels) (i) at the proximal and (ii) distal ends of the tibia reveal extensive osteolytic bone degradation (osteolytic damage highlighted with white arrows) near the injection site. Micro-CT images scanned using a Microphotonics Skyscan 1172 CT at a 6μm resolution and reconstructed using ImageVis3D imaging software.

Mentions: To identify whether 4 T1.2 luc3 cells are osteolytic by comparison to the commercially available 4 T1 luc2 cells, each of the cell lines were injected into the tibia of 6-week old female Balb/c mice and analyzed for bone integrity using micro-CT. Whole body and ex vivo BLI analysis of mice injected with 4 T1.2 luc3 and 4 T1 luc2 cells verified the presence of tumor cells 18 days after injection into the tibia (Figure 7A). Micro-CT analysis showed significant osteolytic activity of both cell lines near the injection site at the proximal tibia (Figure 7B-i) and normal bone at the distal tibia (Figure 7B-ii). 66c14 luc2 cells were not analyzed for osteolytic potential in this study. Overall, these results suggest that 4 T1.2 luc3 cells are a useful model for studying osteolytic bone metastasis originating from a primary breast tumor.


Novel mouse mammary cell lines for in vivo bioluminescence imaging (BLI) of bone metastasis.

Bolin C, Sutherland C, Tawara K, Moselhy J, Jorcyk CL - Biol Proced Online (2012)

4T1.2 luc3 cells are osteolytic in vivo. (A)  1 × 104 4 T1.2 luc3 (left) or 4 T1 luc2 cells (right) were injected into the proximal end of the left tibia of a 6-week old female Balb/c mice and visualized after i.p. injection of 150 mg/kg D-Luciferin using an IVIS Spectrum BLI system (day 18 after tumor cell injection). Whole body ventral view shows luciferase activity from tumor cells in the left tibia, validated with ex-vivo imaging of the tibia. (B) micro-CT images of cross sectional slices (4000 × 4000 pixels) (i) at the proximal and (ii) distal ends of the tibia reveal extensive osteolytic bone degradation (osteolytic damage highlighted with white arrows) near the injection site. Micro-CT images scanned using a Microphotonics Skyscan 1172 CT at a 6μm resolution and reconstructed using ImageVis3D imaging software.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC3473320&req=5

Figure 7: 4T1.2 luc3 cells are osteolytic in vivo. (A) 1 × 104 4 T1.2 luc3 (left) or 4 T1 luc2 cells (right) were injected into the proximal end of the left tibia of a 6-week old female Balb/c mice and visualized after i.p. injection of 150 mg/kg D-Luciferin using an IVIS Spectrum BLI system (day 18 after tumor cell injection). Whole body ventral view shows luciferase activity from tumor cells in the left tibia, validated with ex-vivo imaging of the tibia. (B) micro-CT images of cross sectional slices (4000 × 4000 pixels) (i) at the proximal and (ii) distal ends of the tibia reveal extensive osteolytic bone degradation (osteolytic damage highlighted with white arrows) near the injection site. Micro-CT images scanned using a Microphotonics Skyscan 1172 CT at a 6μm resolution and reconstructed using ImageVis3D imaging software.
Mentions: To identify whether 4 T1.2 luc3 cells are osteolytic by comparison to the commercially available 4 T1 luc2 cells, each of the cell lines were injected into the tibia of 6-week old female Balb/c mice and analyzed for bone integrity using micro-CT. Whole body and ex vivo BLI analysis of mice injected with 4 T1.2 luc3 and 4 T1 luc2 cells verified the presence of tumor cells 18 days after injection into the tibia (Figure 7A). Micro-CT analysis showed significant osteolytic activity of both cell lines near the injection site at the proximal tibia (Figure 7B-i) and normal bone at the distal tibia (Figure 7B-ii). 66c14 luc2 cells were not analyzed for osteolytic potential in this study. Overall, these results suggest that 4 T1.2 luc3 cells are a useful model for studying osteolytic bone metastasis originating from a primary breast tumor.

Bottom Line: The 4 T1.2 luc3 cell line was found to closely model the sites of metastases seen in human patients including lung, liver, and bone.High osteolytic activity of the 4 T1.2 luc3 cells in vivo in the bone microenvironment was also detected.The engineered 4 T1.2 luc3 and 66c14 luc2 cell lines described in this study are valuable tools for studying the cellular events moderating the metastasis of breast tumor cells to bone.

View Article: PubMed Central - HTML - PubMed

Affiliation: Department of Biological Sciences, Boise State University, Boise, ID, USA. cjorcyk@boisestate.edu.

ABSTRACT

Background: Tumor cell lines that can be tracked in vivo during tumorigenesis and metastasis provide vital tools for studying the specific cellular mechanisms that mediate these processes as well as investigating therapeutic targets to inhibit them. The goal of this study was to engineer imageable mouse mammary tumor cell lines with discrete propensities to metastasize to bone in vivo. Two novel luciferase expressing cell lines were developed and characterized for use in the study of breast cancer metastasis to bone in a syngeneic mouse model.

Results: The 4 T1.2 luc3 and 66c14 luc2 cell lines were shown to have high levels of bioluminescence intensity in vitro and in vivo after orthotopic injection into mouse mammary fat pads. The 4 T1.2 luc3 cell line was found to closely model the sites of metastases seen in human patients including lung, liver, and bone. Specifically, 4 T1.2 luc3 cells demonstrated a high incidence of metastasis to spine, with an ex-vivo BLI intensity three orders of magnitude above the commercially available 4 T1 luc2 cells. 66c14 luc2 cells also demonstrated metastasis to spine, which was lower than that of 4 T1.2 luc3 cells but higher than 4 T1 luc2 cells, in addition to previously unreported metastases in the liver. High osteolytic activity of the 4 T1.2 luc3 cells in vivo in the bone microenvironment was also detected.

Conclusions: The engineered 4 T1.2 luc3 and 66c14 luc2 cell lines described in this study are valuable tools for studying the cellular events moderating the metastasis of breast tumor cells to bone.

No MeSH data available.


Related in: MedlinePlus