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| Status |
Public on Apr 01, 2011 |
| Title |
Gene expression profiling of IGROV1 cells after in vitro treatment with ascitic fluid from human ovarian cancer bearing mice |
| Organism |
Homo sapiens |
| Experiment type |
Expression profiling by array
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| Summary |
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.
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| Overall design |
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.
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| Contributor(s) |
De Cecco L, Balsari A, Sommariva M |
| Citation(s) |
21878529 |
| Submission date |
Aug 05, 2010 |
| Last update date |
Aug 16, 2018 |
| Contact name |
Loris De Cecco |
| E-mail(s) |
loris.dececco@istitutotumori.mi.it
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| Organization name |
IRCSS Istituto Nazionale Tumori
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| Street address |
via Venezian 1
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| City |
Milan |
| ZIP/Postal code |
20133 |
| Country |
Italy |
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| Platforms (1) |
| GPL6947 |
Illumina HumanHT-12 V3.0 expression beadchip |
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| Samples (6)
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| This SubSeries is part of SuperSeries: |
| GSE23491 |
Gene expression profile of IGROV1 cells after treatment of saline or CpG-ODN |
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| Relations |
| BioProject |
PRJNA133329 |
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