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IGROV1 miRNA expression analysis after in vivo locoregional treatment with CpG-ODN

ABSTRACT: We reported that peri-tumoral CpG-ODN treatment, probably activating TLR9-expressing cells present in the tumor microenvironment, sensitized cancer cells to DNA-damaging chemotherapy (Cancer Res 2011 Oct 15;71(20):6382-90). Here, we investigated whether this treatment induces a modulation of miRNAs and their involvement in chemotherapy sensitivity. Twenty miRNAs were found differentially expressed in tumors from CpG-ODN-treated mice versus controls. Evaluation of the role of miR-424, miR-340 and miR-302b on cisplatin sensitivity revealed that ectopic expression of miR-302b (up-modulated in our array) in IGROV1 cells significantly improved cisplatin activity. The identification of miRNAs able to modify sensitivity to chemotherapy treatment will provide an experimental base for its future possible use as a target or tool of specific therapies. IGROV1 human ovarian carcinoma cells were adapted to growth intraperitoneally (i.p.) and maintained by serial i.p. passages of ascitic cells into healthy mice. Mice were injected i.p. with 2.5 x 10^6 ascitic cells in 0.2 ml of saline and treated starting 10-11 days later, when mice showed evident and established ascites, with CpG-ODN [phosphorothioated ODN1826 (5'-TCCATGACGTTCCTGACGTT-3')] delivered i.p. at a dose of 20 µg/mouse for 3 consecutive days, control mice received saline (4 mice/group). 24 hours after the last treatment, ascites-bearing mice were sacrificed. Tumor cells adherent to peritoneal wall were collected and immediately frozen in liquid nitrogen until RNA extraction.

ORGANISM(S): Homo sapiens

SUBMITTER: Loris De Cecco

PROVIDER: E-GEOD-41783 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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Gene expression profiling of IGROV1 cells after in vitro treatment with ascitic fluid from human ovarian cancer bearing mice

Project description:CpG-ODN demonstrated anti-tumor activity in IGROV-1 ascites tumor-bearing mice. To evaluate if soluble molecules present in ascitic fluid, presumably released by innate immune cells via TLR9 stimulation, is directly involved with gene expression modulation a microarray experiment was performed. IGROV-1 human ovarian carcinoma cells were adapted to growth intraperitoneally (i.p.) and maintained by serial i.p. passages of ascitic cells into healthy mice as described. Mice were injected i.p. with 2.5 x 106 ascitic cells in 0.2 ml of saline and treated starting 10-11 days later, when mice showed evident and established ascites, with CpG-ODN [phosphorothioated ODN1826 (5’-TCCATGACGTTCCTGACGTT-3’), TriLink Biotechnologies (San Diego, CA, USA)]. Delivered i.p. at a dose of 20 µg/mouse for 3 consecutive days or 1 time, control mice received saline (3 mice/group). Twenty-four hours after the last treatment with saline or CpG-ODN, ascites-bearing mice were sacrificed by cervical dislocation. Ascitic fluid was collected using a heparinized syringe and the volume recorded. The fluid was transferred to a centrifuge tube maintained on ice. After centrifugation, supernatant was removed and stored at -80°C for in vitro experiments. The ovarian cancer cell line IGROV-1 was obtained from the ATCC (Rockville, MD). Cells were routinely maintained in RPMI medium 1640 (Sigma, St. Louis, MO) supplemented with 10% FCS (Sigma) and 2 mM glutamine (Cambrex, East Rutherford, NJ). Cells were maintained at 37°C in a water-saturated atmosphere of 5% CO2 in air. 1x106 IGROV-1 cells were seeded in 6-well plates and, after seeding, cells were treated with 2 ml of culture medium in the presence of ascites from saline-treated mice or CpG-ODN-treated mice (ratio medium:ascites, 1:1) for 24 hours. At the end of treatment, cells were collected and RNA extracted.

2011-04-01 | E-GEOD-23440 | biostudies-arrayexpress

IGROV1 gene expression analysis after in vivo locoregional treatment with CpG-ODN

Project description:Toll-like receptor 9 synthetic agonist oligodeoxynucleotides expressing CpG motifs are currently evaluated for their anti-tumor activity, mainly in association with DNA-damaging drugs. Microarray expression analyses of genes implicates in DNA repair on tumor cells from mice treated with CpG-OD, revealed a down-regulation in tumor cells. These findings provide the first time evidence that immune cells upon TLR9 engagement can sensitize cancer cells to DNA damaging chemotherapy. IGROV-1 human ovarian carcinoma cells were adapted to growth intraperitoneally (i.p.) and maintained by serial i.p. passages of ascitic cells into healthy mice as described. Mice were injected i.p. with 2.5 x 106 ascitic cells in 0.2 ml of saline and treated starting 10-11 days later, when mice showed evident and established ascites, with CpG-ODN [phosphorothioated ODN1826 (5’-TCCATGACGTTCCTGACGTT-3’), TriLink Biotechnologies (San Diego, CA, USA)]. delivered i.p. at a dose of 20 µg/mouse for 3 consecutive days or 1 time, control mice received saline (4 mice/group). 24 hours after the last treatment with saline or CpG-ODN, ascites-bearing mice were sacrificed by cervical dislocation. Tumor cells adhered to peritoneal wall were collected and extraction immediately frozen in liquid nitrogen until RNA extraction.

2011-04-01 | E-GEOD-23441 | biostudies-arrayexpress

Gene expression profile of IGROV1 cells after treatment of saline or CpG-ODN

Project description:This SuperSeries is composed of the following subset Series: GSE23440: Gene expression profiling of IGROV1 cells after in vitro treatment with ascitic fluid from human ovarian cancer bearing mice GSE23441: IGROV1 gene expression analysis after in vivo locoregional treatment with CpG-ODN GSE23442: Gene expression profile of IGROV1 cells after in vitro treatment with CpG-ODN Refer to individual Series

2011-04-01 | E-GEOD-23491 | biostudies-arrayexpress

IGROV1 miRNA expression analysis after in vivo locoregional treatment with CpG-ODN

Project description:We reported that peri-tumoral CpG-ODN treatment, probably activating TLR9-expressing cells present in the tumor microenvironment, sensitized cancer cells to DNA-damaging chemotherapy (Cancer Res 2011 Oct 15;71(20):6382-90). Here, we investigated whether this treatment induces a modulation of miRNAs and their involvement in chemotherapy sensitivity. Twenty miRNAs were found differentially expressed in tumors from CpG-ODN-treated mice versus controls. Evaluation of the role of miR-424, miR-340 and miR-302b on cisplatin sensitivity revealed that ectopic expression of miR-302b (up-modulated in our array) in IGROV1 cells significantly improved cisplatin activity. The identification of miRNAs able to modify sensitivity to chemotherapy treatment will provide an experimental base for its future possible use as a target or tool of specific therapies.

2013-03-31 | GSE41783 | GEO

Gene expression profiling of IGROV1 cells after in vitro treatment with ascitic fluid from human ovarian cancer bearing mice

2011-04-01 | GSE23440 | GEO

Gene expression profile of IGROV1 cells after in vitro treatment with CpG-ODN

2011-04-01 | GSE23442 | GEO

Effect of malignant ascites before and after therapy on the proteome of ovarian cancer cells

Project description:Ovarian cancer is characterized by transcoelomic metastasis into the peritoneal cavity. The peritoneal malignant ascites is enriched with ovarian cancer cells and a small amount of tumor-associated immune cells which create a unique microenvironment actively contributing to progression of the disease. However, it remains unclear how cancer cells communicate to its local environment under the influence of chemotherapy. To address this issue, we performed LC-MS/MS analyses of several primary cultures of ovarian cancer cells from chemonaive malignant ascites incubated for 3 days with autologous pre- or post-chemotherapy ascitic fluids. Enrichment analysis identified prominent upregulation of proteins associated with pathways of DNA repair and cell cycle regulation in tumor cells incubated with ascitic fluids after chemotherapy. Consistently with these data, ascites after therapy increased the resistance of ovarian cancer cells to cisplatin. These findings demonstrate that under stress condition cancer cells can secrete signaling molecules into the extracellular space and contribute to the emergence of tumor chemoresistance during short time period.

2024-05-29 | PXD045647 | Pride

Transcriptomic profiles of ovarian cancer cells incubated with autologous ascitic fluids from patients before and after chemotherapy

Project description:Ovarian cancer is characterized by transcoelomic metastasis into the peritoneal cavity. The peritoneal malignant ascites is enriched with ovarian cancer cells and a small amount of tumor-associated immune cells which create a unique microenvironment actively contributing to progression of the disease. However, it is remain unclear how chemonaive and post-chemotherapy ovarian cancer ascitic fluids influence on cancer cells. To address this issue, we performed RNAseq analysis of primary cultures of ovarian cancer cells incubated for 3 days in the presence of ascites from the same patients before and after chemotherapy. We found that ascites after therapy causes a significant changes in transcriptomic profiles of cancer cells, and these changes are similar in samples obtained from all patients (n=4). Enrichment analysis of differentially expressed genes in tumor cells incubated with ascites after chemotherapy identified prominent up-regulation of genes associated with DNA repair, mitotic cell cycle regulation, and cell cycle checkpoints. These findings demonstrate how ascitic fluids persisted after chemotherapy can contribute to the emergence of tumor chemoresistance during short time period.

2024-04-24 | GSE241909 | GEO

Gene expression profile of IGROV1 cells after in vitro treatment with CpG-ODN

Project description:CpG-ODN is a potent immuno-stimulatory molecule. In order to exclude a direct effect of murine CpG-ODN on IGROV1 human ovarian cancer cell line a gene expression experiment was performed. The ovarian cancer cell line IGROV-1 was obtained from the ATCC (Rockville, MD). Cells were routinely maintained in RPMI medium 1640 (Sigma, St. Louis, MO) supplemented with 10% FCS (Sigma) and 2 mM glutamine (Cambrex, East Rutherford, NJ). Cells were maintained at 37°C in a water-saturated atmosphere of 5% CO2 in air. 1x106 IGROV-1 cells were seeded in 6-well plates and, after seeding, cells were treated with 10uM of CpG-ODN in culture medium [phosphorothioated ODN1826 (5’-TCCATGACGTTCCTGACGTT-3’), TriLink Biotechnologies (San Diego, CA, USA)] for 24 hours. At the end of treatment, cells were collected and RNA extracted.

2011-04-01 | E-GEOD-23442 | biostudies-arrayexpress

Suppression of TLR9-dependent gene expression in vivo mediated by TTAGGG motifs

Project description:Immunostimulatory CpG ODN trigger an innate immune response characterized by the rapid production of pro-inflammatory cytokines and chemokines, an effect that persists for days to weeks in vivo. Previous studies established that gene up-regulation is maximal 3 hr after CpG ODN treatment of normal mice {Klaschik et al. JLB 2009}. The immunostimulatory activity of CpG ODN is blocked by the addition of immunosuppressive ODN expressing multiple TTAGGG motifs. To clarify the mechanism underlying this suppression, changes in gene expresssion were evaluated by microarray in mice treated with 400 ìg of CpG ODN + 400 ìg of suppressive ODN.

2011-06-01 | GSE22156 | GEO

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