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Genome-wide modulation of gene transcription in ovarian carcinoma cells by a new mithramycin analogue.

Vizcaíno C, Núñez LE, Morís F, Portugal J - PLoS ONE (2014)

Bottom Line: Nanomolar concentrations of DIG-MSK abrogated the expression of genes involved in a variety of cell processes including transcription regulation and tumor development, which resulted in cell death.The effect of DIG-MSK in the control of gene expression by other transcription factors was also explored.DIG-MSK affected several biological processes and molecular functions related to transcription and its cellular regulation in A2780 cells, including transcription factor activity.

View Article: PubMed Central - PubMed

Affiliation: Instituto de Biología Molecular de Barcelona, Consejo Superior de Investigaciones Cientificas, Barcelona, Spain.

ABSTRACT
Ovarian cancer has a poor prognosis due to intrinsic or acquired resistance to some cytotoxic drugs, raising the interest in new DNA-binding agents such as mithramycin analogues as potential chemotherapeutic agents in gynecological cancer. Using a genome-wide approach, we have analyzed gene expression in A2780 human ovarian carcinoma cells treated with the novel mithramycin analogue DIG-MSK (demycarosyl-3D-β-D-digitoxosyl-mithramycin SK) that binds to C+G-rich DNA sequences. Nanomolar concentrations of DIG-MSK abrogated the expression of genes involved in a variety of cell processes including transcription regulation and tumor development, which resulted in cell death. Some of those genes have been associated with cell proliferation and poor prognosis in ovarian cancer. Sp1 transcription factor regulated most of the genes that were down-regulated by the drug, as well as the up-regulation of other genes mainly involved in response to cell stress. The effect of DIG-MSK in the control of gene expression by other transcription factors was also explored. Some of them, such as CREB, E2F and EGR1, also recognize C/G-rich regions in gene promoters, which encompass potential DIG-MSK binding sites. DIG-MSK affected several biological processes and molecular functions related to transcription and its cellular regulation in A2780 cells, including transcription factor activity. This new compound might be a promising drug for the treatment of ovarian cancer.

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Analysis of gene expression in A2780 cells treated with DIG-MSK.(A) Venn diagrams representing genes affected, either up-regulated or down-regulated in microarray analysis by treatments with 8 nM or 80 nM DIG-MSK (1.5-fold changes, p<0.05). Numbers inside the intersections correspond to genes influenced by both drug concentrations. (B) Quantitative real-time PCR measurements of a set of genes selected among those differentially expressed in A2780 cells treated with DIG-MSK. Compared to untreated cells, the expression of all these genes changed significantly upon treatment (p<0.05). Histograms represent mean ± SD from 3 independent experiments (**p<0.01, Student’s t-test comparison between treatments with either 8 nM or 80 nM DIG-MSK).
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pone-0104687-g003: Analysis of gene expression in A2780 cells treated with DIG-MSK.(A) Venn diagrams representing genes affected, either up-regulated or down-regulated in microarray analysis by treatments with 8 nM or 80 nM DIG-MSK (1.5-fold changes, p<0.05). Numbers inside the intersections correspond to genes influenced by both drug concentrations. (B) Quantitative real-time PCR measurements of a set of genes selected among those differentially expressed in A2780 cells treated with DIG-MSK. Compared to untreated cells, the expression of all these genes changed significantly upon treatment (p<0.05). Histograms represent mean ± SD from 3 independent experiments (**p<0.01, Student’s t-test comparison between treatments with either 8 nM or 80 nM DIG-MSK).

Mentions: We analyzed the effects of 8 and 80 nM DIG-MSK on gene expression in A2780 human ovarian cancer cells after 24-h treatments. These sub-lethal concentrations were selected from the analysis of cell proliferation (Fig. 2) and viability by Trypan blue exclusion by living cells. The supporting microarray data have been submitted to the GEO repository (http://www.ncbi.nlm.nih.gov/geo/query/acc.cgi?acc=GSE46926). After 24-h continuous treatments, 8 nM DIG-MSK affected the expression of 667 genes (1.5-fold; p<0.05), of which 160 were down-regulated (Fig. 3A). At the same threshold, 80 nM DIG-MSK affected 4889 transcripts, of which 2503 were down-regulated. Most of the genes showing altered expression were down-regulated by the higher concentration. Besides, 105 genes showed down-regulated gene expression at both concentrations (intersection in the Venn diagrams in Fig. 3A).


Genome-wide modulation of gene transcription in ovarian carcinoma cells by a new mithramycin analogue.

Vizcaíno C, Núñez LE, Morís F, Portugal J - PLoS ONE (2014)

Analysis of gene expression in A2780 cells treated with DIG-MSK.(A) Venn diagrams representing genes affected, either up-regulated or down-regulated in microarray analysis by treatments with 8 nM or 80 nM DIG-MSK (1.5-fold changes, p<0.05). Numbers inside the intersections correspond to genes influenced by both drug concentrations. (B) Quantitative real-time PCR measurements of a set of genes selected among those differentially expressed in A2780 cells treated with DIG-MSK. Compared to untreated cells, the expression of all these genes changed significantly upon treatment (p<0.05). Histograms represent mean ± SD from 3 independent experiments (**p<0.01, Student’s t-test comparison between treatments with either 8 nM or 80 nM DIG-MSK).
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4128730&req=5

pone-0104687-g003: Analysis of gene expression in A2780 cells treated with DIG-MSK.(A) Venn diagrams representing genes affected, either up-regulated or down-regulated in microarray analysis by treatments with 8 nM or 80 nM DIG-MSK (1.5-fold changes, p<0.05). Numbers inside the intersections correspond to genes influenced by both drug concentrations. (B) Quantitative real-time PCR measurements of a set of genes selected among those differentially expressed in A2780 cells treated with DIG-MSK. Compared to untreated cells, the expression of all these genes changed significantly upon treatment (p<0.05). Histograms represent mean ± SD from 3 independent experiments (**p<0.01, Student’s t-test comparison between treatments with either 8 nM or 80 nM DIG-MSK).
Mentions: We analyzed the effects of 8 and 80 nM DIG-MSK on gene expression in A2780 human ovarian cancer cells after 24-h treatments. These sub-lethal concentrations were selected from the analysis of cell proliferation (Fig. 2) and viability by Trypan blue exclusion by living cells. The supporting microarray data have been submitted to the GEO repository (http://www.ncbi.nlm.nih.gov/geo/query/acc.cgi?acc=GSE46926). After 24-h continuous treatments, 8 nM DIG-MSK affected the expression of 667 genes (1.5-fold; p<0.05), of which 160 were down-regulated (Fig. 3A). At the same threshold, 80 nM DIG-MSK affected 4889 transcripts, of which 2503 were down-regulated. Most of the genes showing altered expression were down-regulated by the higher concentration. Besides, 105 genes showed down-regulated gene expression at both concentrations (intersection in the Venn diagrams in Fig. 3A).

Bottom Line: Nanomolar concentrations of DIG-MSK abrogated the expression of genes involved in a variety of cell processes including transcription regulation and tumor development, which resulted in cell death.The effect of DIG-MSK in the control of gene expression by other transcription factors was also explored.DIG-MSK affected several biological processes and molecular functions related to transcription and its cellular regulation in A2780 cells, including transcription factor activity.

View Article: PubMed Central - PubMed

Affiliation: Instituto de Biología Molecular de Barcelona, Consejo Superior de Investigaciones Cientificas, Barcelona, Spain.

ABSTRACT
Ovarian cancer has a poor prognosis due to intrinsic or acquired resistance to some cytotoxic drugs, raising the interest in new DNA-binding agents such as mithramycin analogues as potential chemotherapeutic agents in gynecological cancer. Using a genome-wide approach, we have analyzed gene expression in A2780 human ovarian carcinoma cells treated with the novel mithramycin analogue DIG-MSK (demycarosyl-3D-β-D-digitoxosyl-mithramycin SK) that binds to C+G-rich DNA sequences. Nanomolar concentrations of DIG-MSK abrogated the expression of genes involved in a variety of cell processes including transcription regulation and tumor development, which resulted in cell death. Some of those genes have been associated with cell proliferation and poor prognosis in ovarian cancer. Sp1 transcription factor regulated most of the genes that were down-regulated by the drug, as well as the up-regulation of other genes mainly involved in response to cell stress. The effect of DIG-MSK in the control of gene expression by other transcription factors was also explored. Some of them, such as CREB, E2F and EGR1, also recognize C/G-rich regions in gene promoters, which encompass potential DIG-MSK binding sites. DIG-MSK affected several biological processes and molecular functions related to transcription and its cellular regulation in A2780 cells, including transcription factor activity. This new compound might be a promising drug for the treatment of ovarian cancer.

Show MeSH
Related in: MedlinePlus