| Budget Amount *help |
¥3,300,000 (Direct Cost: ¥3,300,000)
Fiscal Year 2005: ¥600,000 (Direct Cost: ¥600,000)
Fiscal Year 2004: ¥1,300,000 (Direct Cost: ¥1,300,000)
Fiscal Year 2003: ¥1,400,000 (Direct Cost: ¥1,400,000)
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| Research Abstract |
The discovery of RNA interference (RNAi) in eukaryotic cells is a major breakthrough in the recent progress of the molecular and cellular biology. RNAi machineries exert biological functions in gene regulation, genome defense, chromatin architecture and dynamics. Today, RNAi has had an enormous impact on the development of novel disease models in animals. In addition, small interfering RNAs (siRNAs), the trigger molecules for RNA silencing, are likely to become invaluable tools for the treatment of various diseases.
All of known human myxoid and round cell liposarcomas (MLS/RCLS) are associated with chromosomal translocations. These chromosomal translocations lead to gene fusions that encode chimeric oncoproteins consisting of an N terminus contributed by one of two related genes, TLS (also known as FUS) or EWS, and a C terminus contributed by the CHOP (also called GADD153) gene.
In this project, using the RNA interference method, I show that TLS-CHOP-targeting small interfering RNAs (siRNA) depleted TLS-CHOP protein and caused growth inhibition in vitro. However, we could not detect growth inhibition effects on mouse models. On the other hand, TLS-CHOP works as a tumor specific transcription factor. Therefore, I am trying to identify the novel therapeutic target molecle using microarray analysis.
In addition, I have generated monoclonal antibodies against the unique peptide sequence of TLS/EWS-CHOP oncoproteins. Immunohistochemical analysis using one of the antibodies was more sensitive than the nested RT-PCR for detection of the TLS-CHOP transcripts or its gene products in paraffin-embedded tissue samples. Thus, I expect that the antibody has a great advantage for molecular diagnosis of MLS/RCLS.
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