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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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Related in: MedlinePlus

Chromatin immunoprecipitation analysis of Sp1 binding to the promoters of XIAP, CRABP1, MDK and KCNMA1 genes in A2780 cells in the presence/absence of 80 nM DIG-MSK.ChIP was performed using an anti-Sp1 specific antibody. A DNA fragment that does not contain Sp1-binding sites was also immunoprecipitated as a negative control, as well as an unspecific immunoprecipitation using IgG (Mock). DNA in both the input and in the immunoprecipitated fractions was quantified by qRT-PCR. Data (mean ± SD from 3 independent experiments) are shown as the enrichment of input DNA in the immunoprecipitated fractions (**p<0.01; Student’s t-test).
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pone-0104687-g005: Chromatin immunoprecipitation analysis of Sp1 binding to the promoters of XIAP, CRABP1, MDK and KCNMA1 genes in A2780 cells in the presence/absence of 80 nM DIG-MSK.ChIP was performed using an anti-Sp1 specific antibody. A DNA fragment that does not contain Sp1-binding sites was also immunoprecipitated as a negative control, as well as an unspecific immunoprecipitation using IgG (Mock). DNA in both the input and in the immunoprecipitated fractions was quantified by qRT-PCR. Data (mean ± SD from 3 independent experiments) are shown as the enrichment of input DNA in the immunoprecipitated fractions (**p<0.01; Student’s t-test).

Mentions: Chromatin immunoprecipitation (ChIP) was used to measure Sp1 occupancy at the promoters of XIAP, CRABP1, MDK and KCNMA1 genes, which were repressed by the treatments with 80 nM DIG-MSK. All of them are involved in the development of ovarian carcinoma [6]. A negative control was also included, which consisted of a down-stream region of the housekeeping GAPDH gene that lacks Sp1-binding sites. DIG-MSK decreased Sp1 binding to those promoters, with a superior effect on KCNMA1 (Fig. 5).


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)

Chromatin immunoprecipitation analysis of Sp1 binding to the promoters of XIAP, CRABP1, MDK and KCNMA1 genes in A2780 cells in the presence/absence of 80 nM DIG-MSK.ChIP was performed using an anti-Sp1 specific antibody. A DNA fragment that does not contain Sp1-binding sites was also immunoprecipitated as a negative control, as well as an unspecific immunoprecipitation using IgG (Mock). DNA in both the input and in the immunoprecipitated fractions was quantified by qRT-PCR. Data (mean ± SD from 3 independent experiments) are shown as the enrichment of input DNA in the immunoprecipitated fractions (**p<0.01; Student’s t-test).
© Copyright Policy
Related In: Results  -  Collection

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

pone-0104687-g005: Chromatin immunoprecipitation analysis of Sp1 binding to the promoters of XIAP, CRABP1, MDK and KCNMA1 genes in A2780 cells in the presence/absence of 80 nM DIG-MSK.ChIP was performed using an anti-Sp1 specific antibody. A DNA fragment that does not contain Sp1-binding sites was also immunoprecipitated as a negative control, as well as an unspecific immunoprecipitation using IgG (Mock). DNA in both the input and in the immunoprecipitated fractions was quantified by qRT-PCR. Data (mean ± SD from 3 independent experiments) are shown as the enrichment of input DNA in the immunoprecipitated fractions (**p<0.01; Student’s t-test).
Mentions: Chromatin immunoprecipitation (ChIP) was used to measure Sp1 occupancy at the promoters of XIAP, CRABP1, MDK and KCNMA1 genes, which were repressed by the treatments with 80 nM DIG-MSK. All of them are involved in the development of ovarian carcinoma [6]. A negative control was also included, which consisted of a down-stream region of the housekeeping GAPDH gene that lacks Sp1-binding sites. DIG-MSK decreased Sp1 binding to those promoters, with a superior effect on KCNMA1 (Fig. 5).

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