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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.


Statistical analysis of mRNA levels. (A) Spearman's rank correlation test for the genes of interest and the proportion of total VEGF-A in the form of VEGF189. Differences were considered significant if P<0.05. (B) Survival curves with an optimal cutoff point. Survival was estimated by the Kaplan–Meier method, and groups were compared in log-rank tests. The optimal cutoff point was calculated for each gene and used to distinguish two groups as described in Materials and methods.
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fig5: Statistical analysis of mRNA levels. (A) Spearman's rank correlation test for the genes of interest and the proportion of total VEGF-A in the form of VEGF189. Differences were considered significant if P<0.05. (B) Survival curves with an optimal cutoff point. Survival was estimated by the Kaplan–Meier method, and groups were compared in log-rank tests. The optimal cutoff point was calculated for each gene and used to distinguish two groups as described in Materials and methods.

Mentions: We then carried out Spearman's rank correlation tests to assess the extent to which the expression of the 19 genes identified (18+TNC) was correlated with that of VEGF189/VEGFA ratio (Figure 5A). We found a statistically negative correlation between the expression pattern of VEGF189 and the levels of expression of seven genes: CARS (r=−0.381, P=0.000035), CPNE3 (r=−0.226, P=0.012), GINS2 (r=−0.273, P=0.0027), GPRC5A (r=−0.219, P=0.016), HIST1H2BK (r=−0.338, P=0.00023), TBC1D8 (r=−0.181, P=0.045) and ZDHHC13 (r=−0.270, P=0.003). Levels of VEGF165 expression were found to be correlated with the levels of expression of CTSL-1, GGH, HIST1H2BK, HRASLS, SFT2D2, TNC and MMP1 (data not shown).


MDA-MB-231 breast cancer cells overexpressing single VEGF isoforms display distinct colonisation characteristics
Statistical analysis of mRNA levels. (A) Spearman's rank correlation test for the genes of interest and the proportion of total VEGF-A in the form of VEGF189. Differences were considered significant if P<0.05. (B) Survival curves with an optimal cutoff point. Survival was estimated by the Kaplan–Meier method, and groups were compared in log-rank tests. The optimal cutoff point was calculated for each gene and used to distinguish two groups as described in Materials and methods.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4559830&req=5

fig5: Statistical analysis of mRNA levels. (A) Spearman's rank correlation test for the genes of interest and the proportion of total VEGF-A in the form of VEGF189. Differences were considered significant if P<0.05. (B) Survival curves with an optimal cutoff point. Survival was estimated by the Kaplan–Meier method, and groups were compared in log-rank tests. The optimal cutoff point was calculated for each gene and used to distinguish two groups as described in Materials and methods.
Mentions: We then carried out Spearman's rank correlation tests to assess the extent to which the expression of the 19 genes identified (18+TNC) was correlated with that of VEGF189/VEGFA ratio (Figure 5A). We found a statistically negative correlation between the expression pattern of VEGF189 and the levels of expression of seven genes: CARS (r=−0.381, P=0.000035), CPNE3 (r=−0.226, P=0.012), GINS2 (r=−0.273, P=0.0027), GPRC5A (r=−0.219, P=0.016), HIST1H2BK (r=−0.338, P=0.00023), TBC1D8 (r=−0.181, P=0.045) and ZDHHC13 (r=−0.270, P=0.003). Levels of VEGF165 expression were found to be correlated with the levels of expression of CTSL-1, GGH, HIST1H2BK, HRASLS, SFT2D2, TNC and MMP1 (data not shown).

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.