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Pleiotropic Anti-Angiogenic and Anti-Oncogenic Activities of the Novel Mithralog Demycarosyl-3D-ß-D-Digitoxosyl-Mithramycin SK (EC-8042).

Fernández-Guizán A, López-Soto A, Acebes-Huerta A, Huergo-Zapico L, Villa-Álvarez M, Núñez LE, Morís F, Gonzalez S - PLoS ONE (2015)

Bottom Line: Accordingly, DIG-MSK also exhibited potent anti-angiogenic activity on microvascular endothelial cells.Likewise, it significantly inhibited the gene expression of VEGFR1, VEGFR2, FGFR, PDGFB and PDGFRA and, additionally, it induced the expression of the anti-angiogenic factors angiostatin and tunstatin.These effects correlated with a pro-apoptotic effect on proliferating microvascular endothelial cells and the inhibition of the formation of endothelial capillary structures.

View Article: PubMed Central - PubMed

Affiliation: Department of Functional Biology, IUOPA, Universidad de Oviedo, Oviedo, Spain.

ABSTRACT
Demycarosyl-3D-ß-D-digitoxosyl-mithramycin SK (DIG-MSK) is a recently isolated analogue of mithramycin A (MTA) that showed differences with MTA in the DNA binding strength and selectivity. These differences correlated with a better therapeutic index and less toxicity in animal studies. Herein, we show that DIG-MSK displays a potent anti-tumor activity against different types of cancer cell lines, ovarian tumor cells being particularly sensitive to this drug. Of relevance, DIG-MSK exerts low toxicity on fibroblasts and peripheral blood mononuclear cells, this toxicity being significantly lower than that of MTA. In correlation with its antitumor activity, DIG-MSK strongly inhibited Sp1-mediated transcription and endogenous Sp1 mRNA expression, which correlated with the inhibition of the expression of key Sp1-regulated genes involved in tumorigenesis, including VEGFA, BCL2L1 (Bcl-XL), hTERT, BRCA2, MYC and SRC in several ovarian cells. Significantly, DIG-MSK was a stronger inhibitor of VEGFA expression than MTA. Accordingly, DIG-MSK also exhibited potent anti-angiogenic activity on microvascular endothelial cells. Likewise, it significantly inhibited the gene expression of VEGFR1, VEGFR2, FGFR, PDGFB and PDGFRA and, additionally, it induced the expression of the anti-angiogenic factors angiostatin and tunstatin. These effects correlated with a pro-apoptotic effect on proliferating microvascular endothelial cells and the inhibition of the formation of endothelial capillary structures. Overall, the pleiotropic activity of DIG-MSK in inhibiting key oncogenic and angiogenic pathways, together with its low toxicity profile, highlight the therapeutic potential of this new drug.

No MeSH data available.


Related in: MedlinePlus

Analysis of the expression of key anti-angiogenic genes in ECs treated with MTA or DIG-MSK.qRT-PCR analysis of the expression of several key anti-angiogenic genes in the presence of 200 nM MTA or DIG-MSK compared to untreated ECs. Data represent the mean ± SEM of Ct values obtained from at least three qRT-PCR independent experiments made by duplicate in HUVEC (a) or HMEC-1 (b) endothelial cells. The relative mRNA expression was obtained by comparison of the expression profiles of untreated cells (DMSO) versus treated ones (*p<0.05; Mann-Whitney U test).
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pone.0140786.g008: Analysis of the expression of key anti-angiogenic genes in ECs treated with MTA or DIG-MSK.qRT-PCR analysis of the expression of several key anti-angiogenic genes in the presence of 200 nM MTA or DIG-MSK compared to untreated ECs. Data represent the mean ± SEM of Ct values obtained from at least three qRT-PCR independent experiments made by duplicate in HUVEC (a) or HMEC-1 (b) endothelial cells. The relative mRNA expression was obtained by comparison of the expression profiles of untreated cells (DMSO) versus treated ones (*p<0.05; Mann-Whitney U test).

Mentions: To gain further insight into the mechanism underpinning the suppression of capillary formation by DIG-MSK, the impact of this drug on the transcription of key angiogenic genes (VEGFA, VEGFB, VEGFC, VEGFD, VEGFR1, VEGFR2, VEGFR3, FGF, FGFR, PDGFA, PDGFB, PDGFRA, PDGFRB, EGF and HIF1A) in ECs was analyzed by qRT-PCR. As shown in Fig 7A, a significant reduction in VEGFR1 and VEGFR2 gene expression in HUVEC cells (ranging from 40 to 50% of inhibition). A down-regulation of VEGFR2, FGFR and PDGFRA gene expression was observed with both analogues in HMEC-1 cells. DIG-MSK also inhibited VEGFR1 and PDGFB expression in these cells (Fig 7A and 7B). Contrarily, MTA significantly up-regulated the angiogenic factor PDGFA in HUVEC cells. Similarly, their effect on the transcript levels of key anti-angiogenic genes encoding angiostatin, tunstatin, endostatin and thrombospondin-1 was analyzed (Fig 8). Angiostatin gene expression was up-regulated by both analogues and tunstatin was induced by DIG-MSK in HUVEC cells. Conversely, a reduction of endostatin expression was observed in HUVEC cells treated with both drugs.


Pleiotropic Anti-Angiogenic and Anti-Oncogenic Activities of the Novel Mithralog Demycarosyl-3D-ß-D-Digitoxosyl-Mithramycin SK (EC-8042).

Fernández-Guizán A, López-Soto A, Acebes-Huerta A, Huergo-Zapico L, Villa-Álvarez M, Núñez LE, Morís F, Gonzalez S - PLoS ONE (2015)

Analysis of the expression of key anti-angiogenic genes in ECs treated with MTA or DIG-MSK.qRT-PCR analysis of the expression of several key anti-angiogenic genes in the presence of 200 nM MTA or DIG-MSK compared to untreated ECs. Data represent the mean ± SEM of Ct values obtained from at least three qRT-PCR independent experiments made by duplicate in HUVEC (a) or HMEC-1 (b) endothelial cells. The relative mRNA expression was obtained by comparison of the expression profiles of untreated cells (DMSO) versus treated ones (*p<0.05; Mann-Whitney U test).
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4633274&req=5

pone.0140786.g008: Analysis of the expression of key anti-angiogenic genes in ECs treated with MTA or DIG-MSK.qRT-PCR analysis of the expression of several key anti-angiogenic genes in the presence of 200 nM MTA or DIG-MSK compared to untreated ECs. Data represent the mean ± SEM of Ct values obtained from at least three qRT-PCR independent experiments made by duplicate in HUVEC (a) or HMEC-1 (b) endothelial cells. The relative mRNA expression was obtained by comparison of the expression profiles of untreated cells (DMSO) versus treated ones (*p<0.05; Mann-Whitney U test).
Mentions: To gain further insight into the mechanism underpinning the suppression of capillary formation by DIG-MSK, the impact of this drug on the transcription of key angiogenic genes (VEGFA, VEGFB, VEGFC, VEGFD, VEGFR1, VEGFR2, VEGFR3, FGF, FGFR, PDGFA, PDGFB, PDGFRA, PDGFRB, EGF and HIF1A) in ECs was analyzed by qRT-PCR. As shown in Fig 7A, a significant reduction in VEGFR1 and VEGFR2 gene expression in HUVEC cells (ranging from 40 to 50% of inhibition). A down-regulation of VEGFR2, FGFR and PDGFRA gene expression was observed with both analogues in HMEC-1 cells. DIG-MSK also inhibited VEGFR1 and PDGFB expression in these cells (Fig 7A and 7B). Contrarily, MTA significantly up-regulated the angiogenic factor PDGFA in HUVEC cells. Similarly, their effect on the transcript levels of key anti-angiogenic genes encoding angiostatin, tunstatin, endostatin and thrombospondin-1 was analyzed (Fig 8). Angiostatin gene expression was up-regulated by both analogues and tunstatin was induced by DIG-MSK in HUVEC cells. Conversely, a reduction of endostatin expression was observed in HUVEC cells treated with both drugs.

Bottom Line: Accordingly, DIG-MSK also exhibited potent anti-angiogenic activity on microvascular endothelial cells.Likewise, it significantly inhibited the gene expression of VEGFR1, VEGFR2, FGFR, PDGFB and PDGFRA and, additionally, it induced the expression of the anti-angiogenic factors angiostatin and tunstatin.These effects correlated with a pro-apoptotic effect on proliferating microvascular endothelial cells and the inhibition of the formation of endothelial capillary structures.

View Article: PubMed Central - PubMed

Affiliation: Department of Functional Biology, IUOPA, Universidad de Oviedo, Oviedo, Spain.

ABSTRACT
Demycarosyl-3D-ß-D-digitoxosyl-mithramycin SK (DIG-MSK) is a recently isolated analogue of mithramycin A (MTA) that showed differences with MTA in the DNA binding strength and selectivity. These differences correlated with a better therapeutic index and less toxicity in animal studies. Herein, we show that DIG-MSK displays a potent anti-tumor activity against different types of cancer cell lines, ovarian tumor cells being particularly sensitive to this drug. Of relevance, DIG-MSK exerts low toxicity on fibroblasts and peripheral blood mononuclear cells, this toxicity being significantly lower than that of MTA. In correlation with its antitumor activity, DIG-MSK strongly inhibited Sp1-mediated transcription and endogenous Sp1 mRNA expression, which correlated with the inhibition of the expression of key Sp1-regulated genes involved in tumorigenesis, including VEGFA, BCL2L1 (Bcl-XL), hTERT, BRCA2, MYC and SRC in several ovarian cells. Significantly, DIG-MSK was a stronger inhibitor of VEGFA expression than MTA. Accordingly, DIG-MSK also exhibited potent anti-angiogenic activity on microvascular endothelial cells. Likewise, it significantly inhibited the gene expression of VEGFR1, VEGFR2, FGFR, PDGFB and PDGFRA and, additionally, it induced the expression of the anti-angiogenic factors angiostatin and tunstatin. These effects correlated with a pro-apoptotic effect on proliferating microvascular endothelial cells and the inhibition of the formation of endothelial capillary structures. Overall, the pleiotropic activity of DIG-MSK in inhibiting key oncogenic and angiogenic pathways, together with its low toxicity profile, highlight the therapeutic potential of this new drug.

No MeSH data available.


Related in: MedlinePlus