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| Status |
Public on Nov 27, 2012 |
| Title |
Downregulation of the HOPX gene decreases metastatic activity in a chicken sarcoma cell line model and identifies genes associated with metastasis |
| Organism |
Gallus gallus |
| Experiment type |
Expression profiling by array
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| Summary |
Metastatic progression is the leading cause of cancer mortality yet we have an incomplete view of the genetic events governing this process. An investigation was undertaken to explore the role of homeodemain only protein X (HOPX) in metastatic propensity and to identify other genes that may participate in metastasis development. The transcription factor HOPX was assessed for its possible involvement in metastasis formation using a knock-down induced by plasmid-delivered shRNAs. We used our original model system of chicken v-src-transformed tumour cell line PR9692 and its subclone (PR9692-E9) that have lost the ability to induce metastases after inoculation into syngeneic chickens without any significant change in primary tumour formation. We found that also a PR9692 cell line with decreased expression of HOPX gene (PR9692-shHOPX) lost its metastatic capacity in vivo (in chickens) and displayed a reduced cell migration in vitro. We compared the gene expression profiles of control (PR9692-shMOCK) and PR9692-shHOPX cells using oligonucleotide microarrays, assuming that genes with differential expression might be associated with metastasis. The data were compared with a previous study showing differences in gene expression between the PR9692 and PR9692-E9 cells. Bioinformatics was applied to identify gene expression patterns associated with metastasis. 234 genes were identified to show at least 2-fold change in both pairs of cell lines. The results were validated with real-time quantitative RT-PCR and the differential expression was confirmed for several genes. We were also able to demonstrate a significant change at protein level in case of three selected genes (NCAM, FOXG1, ITGA4). shRNA mediated knockdown of one of the identified HOPX regulated genes (integrin alpha 4) in the PR9692 cell line itself showed a marked inhibition of metastasis formation.
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| Overall design |
Comparison of expression profiles obtained from metastatic PR9692-shMOCK and non-metastatic PR9692-shHOPX cells. Two biological replicates were analyzed for each group.
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| Contributor(s) |
Kovarova D, Plachy J, Trejbalova K, Cermak V, Kosla J, Hejnar J |
| Citation(s) |
23938949 |
| Submission date |
Nov 26, 2012 |
| Last update date |
Nov 06, 2013 |
| Contact name |
Vladimir Cermak |
| E-mail(s) |
cermak@img.cas.cz
|
| Organization name |
Institute of Molecular Genetics, Czech Academy of Sciences
|
| Lab |
Molecular Virology
|
| Street address |
Videnska 1083
|
| City |
Prague |
| ZIP/Postal code |
14220 |
| Country |
Czech Republic |
| |
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| Platforms (1) |
| GPL3213 |
[Chicken] Affymetrix Chicken Genome Array |
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| Samples (4)
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| GSM1041352 |
PR9692 expressing control shRNA, biological rep1 |
| GSM1041353 |
PR9692 expressing control shRNA, biological rep2 |
| GSM1041354 |
PR9692 expressing anti-HOPX shRNA, biological rep1 |
| GSM1041355 |
PR9692 expressing anti-HOPX shRNA, biological rep2 |
|
| Relations |
| BioProject |
PRJNA182186 |
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