| Budget Amount *help |
¥4,010,000 (Direct Cost: ¥3,500,000、Indirect Cost: ¥510,000)
Fiscal Year 2007: ¥2,210,000 (Direct Cost: ¥1,700,000、Indirect Cost: ¥510,000)
Fiscal Year 2006: ¥1,800,000 (Direct Cost: ¥1,800,000)
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| Research Abstract |
During culture, a-catenin-deficient DLD1 cells detach from the substrate, float in the medium, and then reattach to the substratum. Expression of α-catenin suppress the detachment of the cells. We did following experiments in order to know the reason why the expression of α-catenin results in suppression of the cell detachment. 1) Although DNA micro array analysis revealed differential expression of some matrix metalloproteinase genes, real-time PCR analysis, however, did not confirm the results. 2) Addition of inhibitors for the metalloproteinases did not suppress the detachment of the cells. 3) The expression of integrins or the extracellular matrix components did not changed after α-catenin expression. Therefore we thought that the α-catenin induced changes inside of the cells instead of outside of the cells. The actin filaments are involved not only in the maintenance of the cell morphology but also in cell motility and growth. 4) Analysis with reagents that affect actin cytoskeleton suggested possible involvement of RhoGTPase. Therefore we introduced the dominant-negative Rho, the constitutively active Rho, or the dominant-negative form of RhoGAP to find the role of Rho. The expression of the constructs, however, did not suppress the detachment of the cells. 5) Next, we identified the region of α-catenin that is required for the suppression of the cell detachment. We found that the amino-terminal region of 400 amino acid residues or the carboxy-terminal region of 300 amino acid residues is required for the activity. The research along this line is now in progress.
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