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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

Metastasis of 66c14 luc2, 4 T1.2 luc3, and 4 T1 luc2 cells in vivo . Representative (A) whole animal and (B)ex vivo  organs from mice injected with the 66c14 luc2 (left), 4 T1.2 luc3 (middle), or commercially available 4 T1 luc2 (right) cells, 4–5 weeks after injection are shown. Primary tumor bioluminescence intensities were visualized in animals injected with all cell types in both dorsal and ventral views. Bioluminescence was also detectable in the thoracic region of the 66c14 luc2 and 4 T1.2 luc3 animals correlating with ex vivo  bioluminescence from metastatic lesions detected in lung and spine. No bioluminescence representing metastatic lesions was detected in mice injected with 4 T1 luc2 cells. The bioluminescence intensity of the primary tumor from the 4 T1 luc2 mouse represented was very high and likely prohibited secondary sites of metastasis from being visualized. The BLI scale next to the whole body image of the 4 T1 luc2 injected mouse (highlighted in red) reflects this difference. All other images (highlighted in yellow) are presented with a consistent BLI scale. Overall, the 4 T1.2 luc3 cells showed the highest levels of BLI from metastases both in vivo  and ex-vivo.
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Figure 5: Metastasis of 66c14 luc2, 4 T1.2 luc3, and 4 T1 luc2 cells in vivo . Representative (A) whole animal and (B)ex vivo organs from mice injected with the 66c14 luc2 (left), 4 T1.2 luc3 (middle), or commercially available 4 T1 luc2 (right) cells, 4–5 weeks after injection are shown. Primary tumor bioluminescence intensities were visualized in animals injected with all cell types in both dorsal and ventral views. Bioluminescence was also detectable in the thoracic region of the 66c14 luc2 and 4 T1.2 luc3 animals correlating with ex vivo bioluminescence from metastatic lesions detected in lung and spine. No bioluminescence representing metastatic lesions was detected in mice injected with 4 T1 luc2 cells. The bioluminescence intensity of the primary tumor from the 4 T1 luc2 mouse represented was very high and likely prohibited secondary sites of metastasis from being visualized. The BLI scale next to the whole body image of the 4 T1 luc2 injected mouse (highlighted in red) reflects this difference. All other images (highlighted in yellow) are presented with a consistent BLI scale. Overall, the 4 T1.2 luc3 cells showed the highest levels of BLI from metastases both in vivo and ex-vivo.

Mentions: In the dorsal and ventral whole body BLI images, the 66c14 luc2 and 4 T1.2 luc3 cells were visible in the primary tumors and numerous secondary metastatic regions in vivo using a BLI intensity imaging scale on the order of 106 photons/sec/cm2 (highlighted with a yellow box in Figure 5A). On the other hand, in whole body BLI images of mice injected with the commercially available 4 T1 luc2 cells only primary tumors were visible. The high bioluminescence produced by the 4 T1 luc2 primary tumor was imaged using a scale on the order of 108 photons/sec/cm2 (highlighted with a red box in Figure 5A), and likely prohibited secondary sites of metastases to be visualized.


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)

Metastasis of 66c14 luc2, 4 T1.2 luc3, and 4 T1 luc2 cells in vivo . Representative (A) whole animal and (B)ex vivo  organs from mice injected with the 66c14 luc2 (left), 4 T1.2 luc3 (middle), or commercially available 4 T1 luc2 (right) cells, 4–5 weeks after injection are shown. Primary tumor bioluminescence intensities were visualized in animals injected with all cell types in both dorsal and ventral views. Bioluminescence was also detectable in the thoracic region of the 66c14 luc2 and 4 T1.2 luc3 animals correlating with ex vivo  bioluminescence from metastatic lesions detected in lung and spine. No bioluminescence representing metastatic lesions was detected in mice injected with 4 T1 luc2 cells. The bioluminescence intensity of the primary tumor from the 4 T1 luc2 mouse represented was very high and likely prohibited secondary sites of metastasis from being visualized. The BLI scale next to the whole body image of the 4 T1 luc2 injected mouse (highlighted in red) reflects this difference. All other images (highlighted in yellow) are presented with a consistent BLI scale. Overall, the 4 T1.2 luc3 cells showed the highest levels of BLI from metastases both in vivo  and ex-vivo.
© Copyright Policy - open-access
Related In: Results  -  Collection

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

Figure 5: Metastasis of 66c14 luc2, 4 T1.2 luc3, and 4 T1 luc2 cells in vivo . Representative (A) whole animal and (B)ex vivo organs from mice injected with the 66c14 luc2 (left), 4 T1.2 luc3 (middle), or commercially available 4 T1 luc2 (right) cells, 4–5 weeks after injection are shown. Primary tumor bioluminescence intensities were visualized in animals injected with all cell types in both dorsal and ventral views. Bioluminescence was also detectable in the thoracic region of the 66c14 luc2 and 4 T1.2 luc3 animals correlating with ex vivo bioluminescence from metastatic lesions detected in lung and spine. No bioluminescence representing metastatic lesions was detected in mice injected with 4 T1 luc2 cells. The bioluminescence intensity of the primary tumor from the 4 T1 luc2 mouse represented was very high and likely prohibited secondary sites of metastasis from being visualized. The BLI scale next to the whole body image of the 4 T1 luc2 injected mouse (highlighted in red) reflects this difference. All other images (highlighted in yellow) are presented with a consistent BLI scale. Overall, the 4 T1.2 luc3 cells showed the highest levels of BLI from metastases both in vivo and ex-vivo.
Mentions: In the dorsal and ventral whole body BLI images, the 66c14 luc2 and 4 T1.2 luc3 cells were visible in the primary tumors and numerous secondary metastatic regions in vivo using a BLI intensity imaging scale on the order of 106 photons/sec/cm2 (highlighted with a yellow box in Figure 5A). On the other hand, in whole body BLI images of mice injected with the commercially available 4 T1 luc2 cells only primary tumors were visible. The high bioluminescence produced by the 4 T1 luc2 primary tumor was imaged using a scale on the order of 108 photons/sec/cm2 (highlighted with a red box in Figure 5A), and likely prohibited secondary sites of metastases to be visualized.

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