Limits...
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

Tumor growth of 66c14 luc and 4 T1.2 luc cells injected orthotopically into Balb/c mice. 66c14 luc and 4 T1.2 luc cell lines were injected into the mammary fat pad of Balb/c mice and tumor volume was measured over 36 days. (A) 66c14 luc2 tumor growth (green/square) was most consistent with parental 66c14Tumor growth (red/circle) and (B) 4 T1.2 luc3 tumor growth (green/square) was most consistent with parental 4 T1.2 (red/circle) tumor growth. Data is expressed as tumor volume (mm3) (mean ± std dev; n = 3).
© Copyright Policy - open-access
Related In: Results  -  Collection

License
getmorefigures.php?uid=PMC3473320&req=5

Figure 2: Tumor growth of 66c14 luc and 4 T1.2 luc cells injected orthotopically into Balb/c mice. 66c14 luc and 4 T1.2 luc cell lines were injected into the mammary fat pad of Balb/c mice and tumor volume was measured over 36 days. (A) 66c14 luc2 tumor growth (green/square) was most consistent with parental 66c14Tumor growth (red/circle) and (B) 4 T1.2 luc3 tumor growth (green/square) was most consistent with parental 4 T1.2 (red/circle) tumor growth. Data is expressed as tumor volume (mm3) (mean ± std dev; n = 3).

Mentions: To confirm that the luc-expressing cell lines grow in vivo in a manner comparable to the parental cells, tumor growth was monitored. 1 × 105 cells (parental 66c14, 66c14 luc2, 66c14 luc3, 66c14 luc4, and 66c14 luc5 or parental 4 T1.2, 4 T1.2 luc1, 4 T1.2 luc2, 4 T1.2 luc3, and 4 T1.2 luc4) were injected into the mammary fat pad of female Balb/c mice (n = 3 per cell line) (Figure 2). Tumor size was measured using calipers twice a week for up to 36 days and expressed as tumor volume (mm3) = (length × width2/2) [11]. Mice injected with 66c14 luc2 cells (Figure 2A; green) displayed a tumor growth pattern similar to that of mice injected with parental 66c14 cells (Figure 2A; red). In vivo analysis of 66c14 luc3 cells demonstrated a small reduction in maximum tumor volume compared to parental 66c14 cells but did not produce bioluminescence in vivo (Figure 2A, Figure 3). Mice injected with 66c14 luc4 or luc5 cells (the cells with high BLI intensities in vitro) had severely reduced tumor growth in vivo (Figure 2A, Figure 3). Injection of 4 T1.2 luc3 cells (the cells with the highest BLI intensity in vitro; Figure 2B; green) resulted in a tumor growth pattern similar to that of parental 4 T1.2 cells (Figure 2B; red). 4 T1.2 luc1, luc2, and luc4 cell injections all resulted in reduced tumor progression in vivo (Figure 2B, Figure 3). Overall, 66c14 luc2 and 4 T1.2 luc3 cell lines demonstrated tumor progression most consistent with parental cell lines.


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)

Tumor growth of 66c14 luc and 4 T1.2 luc cells injected orthotopically into Balb/c mice. 66c14 luc and 4 T1.2 luc cell lines were injected into the mammary fat pad of Balb/c mice and tumor volume was measured over 36 days. (A) 66c14 luc2 tumor growth (green/square) was most consistent with parental 66c14Tumor growth (red/circle) and (B) 4 T1.2 luc3 tumor growth (green/square) was most consistent with parental 4 T1.2 (red/circle) tumor growth. Data is expressed as tumor volume (mm3) (mean ± std dev; n = 3).
© Copyright Policy - open-access
Related In: Results  -  Collection

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

Figure 2: Tumor growth of 66c14 luc and 4 T1.2 luc cells injected orthotopically into Balb/c mice. 66c14 luc and 4 T1.2 luc cell lines were injected into the mammary fat pad of Balb/c mice and tumor volume was measured over 36 days. (A) 66c14 luc2 tumor growth (green/square) was most consistent with parental 66c14Tumor growth (red/circle) and (B) 4 T1.2 luc3 tumor growth (green/square) was most consistent with parental 4 T1.2 (red/circle) tumor growth. Data is expressed as tumor volume (mm3) (mean ± std dev; n = 3).
Mentions: To confirm that the luc-expressing cell lines grow in vivo in a manner comparable to the parental cells, tumor growth was monitored. 1 × 105 cells (parental 66c14, 66c14 luc2, 66c14 luc3, 66c14 luc4, and 66c14 luc5 or parental 4 T1.2, 4 T1.2 luc1, 4 T1.2 luc2, 4 T1.2 luc3, and 4 T1.2 luc4) were injected into the mammary fat pad of female Balb/c mice (n = 3 per cell line) (Figure 2). Tumor size was measured using calipers twice a week for up to 36 days and expressed as tumor volume (mm3) = (length × width2/2) [11]. Mice injected with 66c14 luc2 cells (Figure 2A; green) displayed a tumor growth pattern similar to that of mice injected with parental 66c14 cells (Figure 2A; red). In vivo analysis of 66c14 luc3 cells demonstrated a small reduction in maximum tumor volume compared to parental 66c14 cells but did not produce bioluminescence in vivo (Figure 2A, Figure 3). Mice injected with 66c14 luc4 or luc5 cells (the cells with high BLI intensities in vitro) had severely reduced tumor growth in vivo (Figure 2A, Figure 3). Injection of 4 T1.2 luc3 cells (the cells with the highest BLI intensity in vitro; Figure 2B; green) resulted in a tumor growth pattern similar to that of parental 4 T1.2 cells (Figure 2B; red). 4 T1.2 luc1, luc2, and luc4 cell injections all resulted in reduced tumor progression in vivo (Figure 2B, Figure 3). Overall, 66c14 luc2 and 4 T1.2 luc3 cell lines demonstrated tumor progression most consistent with parental cell lines.

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