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MDA-MB-231 breast cancer cells overexpressing single VEGF isoforms display distinct colonisation characteristics

View Article: PubMed Central - PubMed

ABSTRACT

Background:: Vascular endothelial growth factor (VEGF) is a multifunctional cytokine that has important roles in angiogenesis. Our knowledge of the significance of VEGF isoforms in human cancer remains incomplete.

Methods:: Bioluminescence imaging and transcriptomic analysis were used to study the colonisation capacity of the human breast cancer cells MDA-MB-231 controlling or overexpressing the VEGF165 or VEGF189 isoform (named cV-B, V165-B and V189-B, respectively) in nude mice.

Results:: When injected into the bloodstream, V189-B cells induced less metastasis in the lungs and bone than V165-B and cV-B control cells, consistent with longer survival of these mice and delay in tumour uptake in the mice injected with a V189-B clone. Histological analysis confirmed that there were less αSMA-positive cells in the lungs of the mice injected with V189-B. In vitro V189-B cells decreased both cell invasion and survival. Using transcriptomic analysis, we identified a subset of 18 genes expressed differentially between V189 and V165 cell lines and in 120 human breast tumours. V165 was associated with poor prognosis, whereas V189 was not, suggesting a complex regulation by VEGF isoforms. Our results showed a negative correlation between the expression pattern of VEGF189 and the levels of expression of seven genes that influence metastasis.

Conclusion:: Our findings provide the first evidence that VEGF isoforms have different effects on breast cancer cell line colonisation in vivo.

No MeSH data available.


Related in: MedlinePlus

VEGF189 overexpression in MDA-MB-231 breast cancer cells decreases the number of mice with colonisation sites in vivo and delays colonisation in the bone and lungs. (A) Tumour formation for the various bioluminescent clones injected into mice (n=10). Left panel: number of mice with colonisation sites. Right panel: number of colonisation sites per mouse. (B) Number of mice with colonisation present in the bone and lungs after injection with the various clones (V165, V189 and cV). At the indicated times, the total numbers of mice with metastasis in the bone (legs) and lungs were counted in each group. *PV189vs PC-V and PV165<0.05 and **PV189vs PC-V<0.05. (C) Haematoxylin- and eosin-stained sections of breast cancer tumour in lung tissue (T). Different patterns of colonisation are shown: more or less invasive (middle and right, respectively). Original objective magnification, × 16 (top), × 40 (bottom), scale bars are 500 and 200 μm, respectively.
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fig1: VEGF189 overexpression in MDA-MB-231 breast cancer cells decreases the number of mice with colonisation sites in vivo and delays colonisation in the bone and lungs. (A) Tumour formation for the various bioluminescent clones injected into mice (n=10). Left panel: number of mice with colonisation sites. Right panel: number of colonisation sites per mouse. (B) Number of mice with colonisation present in the bone and lungs after injection with the various clones (V165, V189 and cV). At the indicated times, the total numbers of mice with metastasis in the bone (legs) and lungs were counted in each group. *PV189vs PC-V and PV165<0.05 and **PV189vs PC-V<0.05. (C) Haematoxylin- and eosin-stained sections of breast cancer tumour in lung tissue (T). Different patterns of colonisation are shown: more or less invasive (middle and right, respectively). Original objective magnification, × 16 (top), × 40 (bottom), scale bars are 500 and 200 μm, respectively.

Mentions: Bioluminescent breast cancer cell lines can be used for a rapid and sensitive in vivo detection of cell dissemination throughout the body of immune-deficient mice (Supplementary Data 1A and B; Jenkins et al, 2005; Minn et al, 2005; Abdelkarim et al, 2009). Bioluminescent V189-B and V165-B clones and cV-B control cells were injected into the left heart ventricle of female nude mice (n=10 per group). Fourteen days later, the bioluminescent tumour cells were clearly detectable and their numbers increased over a period of 6–10 weeks. In contrast to cV and V165 clones, V189 resulted in a significantly longer time for tumour colonies to appear in the mice (Figure 1A, left panel). The number of colonisation sites appeared in a smaller proportion after the injection of VEGF189-overexpressing cells than after injection of the controls or VEGF165-overexpressing cells (V189-B vs cV orV165-B) (Figure 1A; right panel; Supplementary Data 2A). After 16 days, only 10% of the mice receiving injections of the V189-B clone had developed tumour colonised sites, against 50% of the mice receiving injections of cV-B and V165-B cells (Figure 1A, left panel). After 38 days, almost all the mice (90–100%) receiving cV-B or V165-B clones injections had developed tumour sites, vs 70% of the mice injected with V189-B clones (Figure 1A, left). V189-B-injected mice had a significantly smaller total number of colonisation sites than those injected with the V165-B and cV-B clones after 16 days (P=0.04) and this difference is lower and only significant for V189 vs cV-B cells (P=0.037) after 38 days of the time course, underlying the delaying effect of VEGF189 overexpression (Figure 1A, right panel). Moreover, the lower colonisation observed with the V189 clone was associated with greater mouse survival (Supplementary Data 2B).


MDA-MB-231 breast cancer cells overexpressing single VEGF isoforms display distinct colonisation characteristics
VEGF189 overexpression in MDA-MB-231 breast cancer cells decreases the number of mice with colonisation sites in vivo and delays colonisation in the bone and lungs. (A) Tumour formation for the various bioluminescent clones injected into mice (n=10). Left panel: number of mice with colonisation sites. Right panel: number of colonisation sites per mouse. (B) Number of mice with colonisation present in the bone and lungs after injection with the various clones (V165, V189 and cV). At the indicated times, the total numbers of mice with metastasis in the bone (legs) and lungs were counted in each group. *PV189vs PC-V and PV165<0.05 and **PV189vs PC-V<0.05. (C) Haematoxylin- and eosin-stained sections of breast cancer tumour in lung tissue (T). Different patterns of colonisation are shown: more or less invasive (middle and right, respectively). Original objective magnification, × 16 (top), × 40 (bottom), scale bars are 500 and 200 μm, respectively.
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fig1: VEGF189 overexpression in MDA-MB-231 breast cancer cells decreases the number of mice with colonisation sites in vivo and delays colonisation in the bone and lungs. (A) Tumour formation for the various bioluminescent clones injected into mice (n=10). Left panel: number of mice with colonisation sites. Right panel: number of colonisation sites per mouse. (B) Number of mice with colonisation present in the bone and lungs after injection with the various clones (V165, V189 and cV). At the indicated times, the total numbers of mice with metastasis in the bone (legs) and lungs were counted in each group. *PV189vs PC-V and PV165<0.05 and **PV189vs PC-V<0.05. (C) Haematoxylin- and eosin-stained sections of breast cancer tumour in lung tissue (T). Different patterns of colonisation are shown: more or less invasive (middle and right, respectively). Original objective magnification, × 16 (top), × 40 (bottom), scale bars are 500 and 200 μm, respectively.
Mentions: Bioluminescent breast cancer cell lines can be used for a rapid and sensitive in vivo detection of cell dissemination throughout the body of immune-deficient mice (Supplementary Data 1A and B; Jenkins et al, 2005; Minn et al, 2005; Abdelkarim et al, 2009). Bioluminescent V189-B and V165-B clones and cV-B control cells were injected into the left heart ventricle of female nude mice (n=10 per group). Fourteen days later, the bioluminescent tumour cells were clearly detectable and their numbers increased over a period of 6–10 weeks. In contrast to cV and V165 clones, V189 resulted in a significantly longer time for tumour colonies to appear in the mice (Figure 1A, left panel). The number of colonisation sites appeared in a smaller proportion after the injection of VEGF189-overexpressing cells than after injection of the controls or VEGF165-overexpressing cells (V189-B vs cV orV165-B) (Figure 1A; right panel; Supplementary Data 2A). After 16 days, only 10% of the mice receiving injections of the V189-B clone had developed tumour colonised sites, against 50% of the mice receiving injections of cV-B and V165-B cells (Figure 1A, left panel). After 38 days, almost all the mice (90–100%) receiving cV-B or V165-B clones injections had developed tumour sites, vs 70% of the mice injected with V189-B clones (Figure 1A, left). V189-B-injected mice had a significantly smaller total number of colonisation sites than those injected with the V165-B and cV-B clones after 16 days (P=0.04) and this difference is lower and only significant for V189 vs cV-B cells (P=0.037) after 38 days of the time course, underlying the delaying effect of VEGF189 overexpression (Figure 1A, right panel). Moreover, the lower colonisation observed with the V189 clone was associated with greater mouse survival (Supplementary Data 2B).

View Article: PubMed Central - PubMed

ABSTRACT

Background:: Vascular endothelial growth factor (VEGF) is a multifunctional cytokine that has important roles in angiogenesis. Our knowledge of the significance of VEGF isoforms in human cancer remains incomplete.

Methods:: Bioluminescence imaging and transcriptomic analysis were used to study the colonisation capacity of the human breast cancer cells MDA-MB-231 controlling or overexpressing the VEGF165 or VEGF189 isoform (named cV-B, V165-B and V189-B, respectively) in nude mice.

Results:: When injected into the bloodstream, V189-B cells induced less metastasis in the lungs and bone than V165-B and cV-B control cells, consistent with longer survival of these mice and delay in tumour uptake in the mice injected with a V189-B clone. Histological analysis confirmed that there were less &alpha;SMA-positive cells in the lungs of the mice injected with V189-B. In vitro V189-B cells decreased both cell invasion and survival. Using transcriptomic analysis, we identified a subset of 18 genes expressed differentially between V189 and V165 cell lines and in 120 human breast tumours. V165 was associated with poor prognosis, whereas V189 was not, suggesting a complex regulation by VEGF isoforms. Our results showed a negative correlation between the expression pattern of VEGF189 and the levels of expression of seven genes that influence metastasis.

Conclusion:: Our findings provide the first evidence that VEGF isoforms have different effects on breast cancer cell line colonisation in vivo.

No MeSH data available.


Related in: MedlinePlus