| Budget Amount *help |
¥3,000,000 (Direct Cost: ¥3,000,000)
Fiscal Year 2004: ¥1,200,000 (Direct Cost: ¥1,200,000)
Fiscal Year 2003: ¥1,800,000 (Direct Cost: ¥1,800,000)
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| Research Abstract |
We have previously established 33 bone marrow stromal cell lines from SV40-T antigen transgenic mice. Of these, 27 clones supported erythroid colony formation, while 6 clones did not. The objective of this study is to identify the molecules which determine these erythroid colony forming activities.
We compared the gene expression profiling by DNA microarray between cell lines which support eaythmpoiesis (E+;TBR9,184,31-2) and cell lines which do not (E-; TBR17,33,511). Among the differentially expressed genes, we selected candidate genes, with the results of quantitative RT-PCR, examined the effect of small interfering RNA (siRNA) and the addition of exogenous proteins on the erythroid colony formation.
Out of 7226 genes examined, 138 and 282 genes were upregulated and downregulated in E+ by 3-fold or more, respectively. We have selected one of the upregulated genes, tenascin-C (TN-C), as a candidate. The expressions of TN-C in E+ were all higher than the three E- cell lines with a mean of 3.6-fold. The number of erythroad colonies in the presence of TN-C siRNA was significantly lower than that of control siRNA in TBR9 (20.7±6.3 versus 4.7±4.8 colonies ; p=0.01) and in TBR184 (13.3±5.3 versus 0.3±0.5;p=0.02). Moreover, the addition of exogenous TN-C enhanced the number of erythmid colonies in TBR184 (13.3±3.5 versus 20.0±2.0;p=0.04) and in TBR31-2 (7.5±3.1 versus 13.5±2.6;p=0.03).
These results suggest that TN-C is responsible for detemining the stromal cell-dependent erythropoiesis.
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