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Status |
Public on Dec 05, 2011 |
Title |
High IGFBP2 Expression Correlates with Tumor Severity in Pediatric Rhabdomyosarcoma. |
Organism |
Homo sapiens |
Experiment type |
Expression profiling by array
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Summary |
Rhabdomyosarcoma (RMS) is the most common childhood sarcoma and is identified as either the embryonal or alveolar (ARMS) subtype. In approximately 75% of cases, ARMSs are characterized by specific chromosomal translocations that involve PAX and FKHR genes. ARMS gene expression signatures vary, depending on the presence or absence of the translocations. Insulin-like growth factor-binding protein 2 (IGFBP2) is strongly overexpressed in translocation-negative RMS. Because IGFBP2 is associated with tumorigenesis, we investigated its functional role in RMS. An analysis of IGFBP2 distribution in RMS cell lines revealed a strong accumulation in the Golgi complex, in which morphological characteristics appeared peculiarly modified. After silencing IGFBP2 expression, our microarray analysis revealed mostly cell cycle and actin cytoskeleton gene modulations. In parallel, IGFBP2-silenced cells showed reduced cell cycle and rates of invasion and decreased seeding in the lungs after tail vein injections in immunodeficient mice. An analysis of IGFBP2 mRNA and protein localization in human tumors showed abnormal protein accumulation in the Golgi complex, mostly in PAX/FKHR-negative RMS. Moreover, an analysis of patients with RMS revealed the presence of conspicuous circulating levels of IGFBP2 proteins in children with highly aggressive RMS tumors. Taken together, our data provide evidence that IGFBP2 contributes to tumor progression and that it could be used as a marker to better classify clinical and biological risks in RMS.
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Overall design |
Three independent silencing experiments on RH36, an embryonal rhabdomyosarcoma cell line, were performed to study the role of IGFBP2 oncogene in this childhood tumor. Each of 3 biological replicates has its own control which is represented by cell culture treated with a non-targeting siRNA (siCONTROL). 3 controls were used to produce a “control pool” that has been used as reference in all microarray experiments. Each microarray experiment consists of a competitive hybridization between test and reference RNA (e.g. silenced cells vs. control), labelled with Cy3 or Cy5. Two microarray experiments were performed for each of three silencing experiment (technical replicate). Each slide was scanned and the data was normalized and analyzed.
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Contributor(s) |
Tombolan L, Casara S, De Pittà C, Orso F, Zin A, Guzzardo V, Romualdi C, Alaggio R, Taverna D, Rosolen A, Lanfranchi G |
Citation(s) |
21924226 |
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Submission date |
Jul 27, 2010 |
Last update date |
Mar 22, 2012 |
Contact name |
Gerolamo Lanfranchi |
E-mail(s) |
stefano.cagnin@unipd.it
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Phone |
+39-0498276219
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Organization name |
University of Padova
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Department |
CRIBI - Biotechnology Center and Biology Department
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Lab |
Functional Genomics Lab
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Street address |
Via U. Bassi, 58/B
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City |
Padova |
ZIP/Postal code |
35131 |
Country |
Italy |
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Platforms (1) |
GPL2136 |
Micro-CRIBI Human Oligo Array (Operon V2.0) |
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Samples (6)
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GSM570808 |
IGFBP2 silencing - experiment 1A |
GSM570809 |
IGFBP2 silencing - experiment 1B (Technical replicate) |
GSM570827 |
IGFBP2 silencing - experiment 2A |
GSM570828 |
IGFBP2 silencing - experiment 2B (Technical replicate) |
GSM570829 |
IGFBP2 silencing - experiment 3A |
GSM570830 |
IGFBP2 silencing - experiment 3B (Technical replicate) |
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Relations |
BioProject |
PRJNA131545 |
Supplementary file |
Size |
Download |
File type/resource |
GSE23196_RAW.tar |
32.9 Mb |
(http)(custom) |
TAR (of TXT) |
Processed data included within Sample table |
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