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Gemcitabine eliminates double minute chromosomes from human ovarian cancer cells.

Yu L, Zhao Y, Quan C, Ji W, Zhu J, Huang Y, Guan R, Sun D, Jin Y, Meng X, Zhang C, Yu Y, Bai J, Sun W, Fu S - PLoS ONE (2013)

Bottom Line: Amplified genes present on the double minute chromosomes are decreased at the DNA level upon gemcitabine treatment.Cells treated with gemcitabine also showed decreased cell growth, colony formation, and invasion.Together, our results suggest that gemcitabine is effective in decreasing double minute chromosomes and this affects the biology of ovarian cancer cells.

View Article: PubMed Central - PubMed

Affiliation: Laboratory of Medical Genetics, Harbin Medical University, Harbin, China.

ABSTRACT
Double minute chromosomes are cytogenetic manifestations of gene amplification frequently seen in cancer cells. Genes amplified on double minute chromosomes include oncogenes and multi-drug resistant genes. These genes encode proteins which contribute to cancer formation, cancer progression, and development of resistance to drugs used in cancer treatment. Elimination of double minute chromosomes, and therefore genes amplified on them, is an effective way to decrease the malignancy of cancer cells. We investigated the effectiveness of a cancer drug, gemcitabine, on the loss of double minute chromosomes from the ovarian cancer cell line UACC-1598. Gemcitabine is able to decrease the number of double minute chromosomes in cells at a 7500X lower concentration than the commonly used cancer drug hydroxyurea. Amplified genes present on the double minute chromosomes are decreased at the DNA level upon gemcitabine treatment. Gemcitabine, even at a low nanomolar concentration, is able to cause DNA damage. The selective incorporation of double minutes chromatin and γ-H2AX signals into micronuclei provides a strong link between DNA damage and the loss of double minute chromosomes from gemcitabine treated cells. Cells treated with gemcitabine also showed decreased cell growth, colony formation, and invasion. Together, our results suggest that gemcitabine is effective in decreasing double minute chromosomes and this affects the biology of ovarian cancer cells.

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The amplification of genes present on DMs is decreased in UACC-1598 cells grown in the presence of HU and GEM by real-time PCR analysis.The amplification level of genes EIF5A2, MCL1, and MYCN was analyzed by real-time PCR. The amplification level of each gene in compound treated cells is compared to control cells (DMSO for HU treated, and Ctrl. for GEM treated), and the mean relative amplification level ± SD is graphed. *denotes a P value of 0.01 to 0.05 and **denotes a P value of 0.001 to 0.01, ***denotes P<0.001.
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pone-0071988-g003: The amplification of genes present on DMs is decreased in UACC-1598 cells grown in the presence of HU and GEM by real-time PCR analysis.The amplification level of genes EIF5A2, MCL1, and MYCN was analyzed by real-time PCR. The amplification level of each gene in compound treated cells is compared to control cells (DMSO for HU treated, and Ctrl. for GEM treated), and the mean relative amplification level ± SD is graphed. *denotes a P value of 0.01 to 0.05 and **denotes a P value of 0.001 to 0.01, ***denotes P<0.001.

Mentions: Furthermore, we also confirmed our results by performing real-time PCR to analyze the amplification of EIF5A2, MYCN, and MCL1 in cells grown in the presence of HU or GEM (Figure 3). We saw a statistically significant decrease in the amplification of EIF5A2 and MCL1 genes when cells were grown in the presence of either HU or GEM. For cells treated with HU, a decrease in relative DNA amplification level of 1.151±0.213 to 0.658±0.092 was observed for EIF5A2, and a decrease in amplification level of 0.860±0.122 to 0.422±0.106 was observed for MCL1. For cells treated with GEM, a decrease in amplification level of 0.930±0.094 to 0.383±0.005 was observed for EIF5A2, and a decrease in amplification level of 1.242±0.343 to 0.388±0.048 was observed for MCL1. However, we did not detect a statistically significant decrease in the amplification of MYCN in either HU or GEM treated cells. Similar results were also acquired for UACC-1598-4 cells, although with obscure change of MCL1 (Figure S1C).


Gemcitabine eliminates double minute chromosomes from human ovarian cancer cells.

Yu L, Zhao Y, Quan C, Ji W, Zhu J, Huang Y, Guan R, Sun D, Jin Y, Meng X, Zhang C, Yu Y, Bai J, Sun W, Fu S - PLoS ONE (2013)

The amplification of genes present on DMs is decreased in UACC-1598 cells grown in the presence of HU and GEM by real-time PCR analysis.The amplification level of genes EIF5A2, MCL1, and MYCN was analyzed by real-time PCR. The amplification level of each gene in compound treated cells is compared to control cells (DMSO for HU treated, and Ctrl. for GEM treated), and the mean relative amplification level ± SD is graphed. *denotes a P value of 0.01 to 0.05 and **denotes a P value of 0.001 to 0.01, ***denotes P<0.001.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC3750019&req=5

pone-0071988-g003: The amplification of genes present on DMs is decreased in UACC-1598 cells grown in the presence of HU and GEM by real-time PCR analysis.The amplification level of genes EIF5A2, MCL1, and MYCN was analyzed by real-time PCR. The amplification level of each gene in compound treated cells is compared to control cells (DMSO for HU treated, and Ctrl. for GEM treated), and the mean relative amplification level ± SD is graphed. *denotes a P value of 0.01 to 0.05 and **denotes a P value of 0.001 to 0.01, ***denotes P<0.001.
Mentions: Furthermore, we also confirmed our results by performing real-time PCR to analyze the amplification of EIF5A2, MYCN, and MCL1 in cells grown in the presence of HU or GEM (Figure 3). We saw a statistically significant decrease in the amplification of EIF5A2 and MCL1 genes when cells were grown in the presence of either HU or GEM. For cells treated with HU, a decrease in relative DNA amplification level of 1.151±0.213 to 0.658±0.092 was observed for EIF5A2, and a decrease in amplification level of 0.860±0.122 to 0.422±0.106 was observed for MCL1. For cells treated with GEM, a decrease in amplification level of 0.930±0.094 to 0.383±0.005 was observed for EIF5A2, and a decrease in amplification level of 1.242±0.343 to 0.388±0.048 was observed for MCL1. However, we did not detect a statistically significant decrease in the amplification of MYCN in either HU or GEM treated cells. Similar results were also acquired for UACC-1598-4 cells, although with obscure change of MCL1 (Figure S1C).

Bottom Line: Amplified genes present on the double minute chromosomes are decreased at the DNA level upon gemcitabine treatment.Cells treated with gemcitabine also showed decreased cell growth, colony formation, and invasion.Together, our results suggest that gemcitabine is effective in decreasing double minute chromosomes and this affects the biology of ovarian cancer cells.

View Article: PubMed Central - PubMed

Affiliation: Laboratory of Medical Genetics, Harbin Medical University, Harbin, China.

ABSTRACT
Double minute chromosomes are cytogenetic manifestations of gene amplification frequently seen in cancer cells. Genes amplified on double minute chromosomes include oncogenes and multi-drug resistant genes. These genes encode proteins which contribute to cancer formation, cancer progression, and development of resistance to drugs used in cancer treatment. Elimination of double minute chromosomes, and therefore genes amplified on them, is an effective way to decrease the malignancy of cancer cells. We investigated the effectiveness of a cancer drug, gemcitabine, on the loss of double minute chromosomes from the ovarian cancer cell line UACC-1598. Gemcitabine is able to decrease the number of double minute chromosomes in cells at a 7500X lower concentration than the commonly used cancer drug hydroxyurea. Amplified genes present on the double minute chromosomes are decreased at the DNA level upon gemcitabine treatment. Gemcitabine, even at a low nanomolar concentration, is able to cause DNA damage. The selective incorporation of double minutes chromatin and γ-H2AX signals into micronuclei provides a strong link between DNA damage and the loss of double minute chromosomes from gemcitabine treated cells. Cells treated with gemcitabine also showed decreased cell growth, colony formation, and invasion. Together, our results suggest that gemcitabine is effective in decreasing double minute chromosomes and this affects the biology of ovarian cancer cells.

Show MeSH
Related in: MedlinePlus