Inhibition of mRNA expression and serum-induced proliferation induced by oligonucleotides directed against rat A-Raf and C-Raf in rat A10 smooth muscle cells
| ISIS | Tm | IC50value | |
|---|---|---|---|
| mRNA expression | Proliferation | ||
| nm | |||
| C-Raf: | |||
| 11061 (active C-Raf ODN) | 60.5 | 55 ± 20 | 216 ± 39 |
| 13490 (1 base mismatch control) | 45.8 | 350 ± 45 | 251 ± 53 |
| 13491 (2 base mismatch control) | 42.2 | >500 | 240 ± 25 |
| 13492 (3 base mismatch control) | 33.8 | >500 | 408 ± 34 |
| 13493 (4 base mismatch control) | 25.0 | >500 | >500 |
| 13494 (5 base mismatch control) | — | >500 | >500 |
| 13495 (6 base mismatch control) | — | >500 | >500 |
| A-Raf: | |||
| 9069 (active A-Raf ODN) | N.D. | 125 ± 25 | 273 ± 34 |
| 11898 (scrambled control) | N.D. | >500 | >750 |
| 11900 (scrambled control) | N.D. | >500 | >750 |
Rat A10 SMCs were treated with the indicated ODNs for 4 hr followed by replacement of media. After a 24-hr incubation, the effects of on mRNA expression and serum-induced proliferation were assessed as described under Experimental Procedures. Thermal melting temperatures (Tm ) were determined for the indicated phosphorothioate ODNs targeted to ISIS 11061 complementary 20-mer RNA as described under Experimental Procedures. See Table 1 for ODN sequences. IC50 values were determined by visual inspection of the normalized graphs. Values shown are mean ± standard error for three to four experiments. N.D., not determined.