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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

Cell cycle distribution and pro-apoptotic effect on microvascular endothelial cells upon exposure to MTA and DIG-MSK.a) HUVEC and HMEC-1 cells treated with MTA (200 nM) or DIG-MSK (200 nM) or DMSO were stained with propidium iodide (PI) and the cell cycle distribution was analyzed by flow cytometry. A representative cytometric profile of HUVEC cells is shown. b) Cell death was analyzed by flow cytometry in ECs (HUVEC and HMEC-1 cells) treated with 200 nM MTA, DIG-MSK or in untreated cells after staining them with Annexin-V and 7-AAD. The bars represent the mean ± SEM of the specific cell death. At least three independent experiments were analyzed (*p<0.05; Mann-Whitney U test).
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pone.0140786.g005: Cell cycle distribution and pro-apoptotic effect on microvascular endothelial cells upon exposure to MTA and DIG-MSK.a) HUVEC and HMEC-1 cells treated with MTA (200 nM) or DIG-MSK (200 nM) or DMSO were stained with propidium iodide (PI) and the cell cycle distribution was analyzed by flow cytometry. A representative cytometric profile of HUVEC cells is shown. b) Cell death was analyzed by flow cytometry in ECs (HUVEC and HMEC-1 cells) treated with 200 nM MTA, DIG-MSK or in untreated cells after staining them with Annexin-V and 7-AAD. The bars represent the mean ± SEM of the specific cell death. At least three independent experiments were analyzed (*p<0.05; Mann-Whitney U test).

Mentions: To further analyze the anti-angiogenic properties of DIG-MSK, its effect on microvascular endothelial cells (ECs) was analyzed. No significant effect was observed on cell cycle progression in HUVEC or HMEC-1 cells (Fig 5A). Likewise, no EC migration was not significantly affected by DIG-MSK, as determined by the wound/scratch-healing assay (not shown). However, DIG-MSK treatment induced significantly lower levels of cell death in HUVEC cells than MTA, with no marked differences observed in HMEC-1 cells (Fig 5B).


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)

Cell cycle distribution and pro-apoptotic effect on microvascular endothelial cells upon exposure to MTA and DIG-MSK.a) HUVEC and HMEC-1 cells treated with MTA (200 nM) or DIG-MSK (200 nM) or DMSO were stained with propidium iodide (PI) and the cell cycle distribution was analyzed by flow cytometry. A representative cytometric profile of HUVEC cells is shown. b) Cell death was analyzed by flow cytometry in ECs (HUVEC and HMEC-1 cells) treated with 200 nM MTA, DIG-MSK or in untreated cells after staining them with Annexin-V and 7-AAD. The bars represent the mean ± SEM of the specific cell death. At least three independent experiments were analyzed (*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.g005: Cell cycle distribution and pro-apoptotic effect on microvascular endothelial cells upon exposure to MTA and DIG-MSK.a) HUVEC and HMEC-1 cells treated with MTA (200 nM) or DIG-MSK (200 nM) or DMSO were stained with propidium iodide (PI) and the cell cycle distribution was analyzed by flow cytometry. A representative cytometric profile of HUVEC cells is shown. b) Cell death was analyzed by flow cytometry in ECs (HUVEC and HMEC-1 cells) treated with 200 nM MTA, DIG-MSK or in untreated cells after staining them with Annexin-V and 7-AAD. The bars represent the mean ± SEM of the specific cell death. At least three independent experiments were analyzed (*p<0.05; Mann-Whitney U test).
Mentions: To further analyze the anti-angiogenic properties of DIG-MSK, its effect on microvascular endothelial cells (ECs) was analyzed. No significant effect was observed on cell cycle progression in HUVEC or HMEC-1 cells (Fig 5A). Likewise, no EC migration was not significantly affected by DIG-MSK, as determined by the wound/scratch-healing assay (not shown). However, DIG-MSK treatment induced significantly lower levels of cell death in HUVEC cells than MTA, with no marked differences observed in HMEC-1 cells (Fig 5B).

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