FIG. 8.

EGF-induced tumorigenicity in nude mice and inhibition of growth on soft agar by antisense oligonucleotides. (A) A549 cells (5 × 10⁶) were injected into nude mice and analyzed for their ability to develop tumors in animals treated daily with saline solution (control) (open bar) or 100, 200, or 300 μg of rhEGF/kg (solid bars). After 8 days, visible tumors were scored as the ratio of the number of tumors over the number of sites injected (indicated above the bars). The asterisk indicates that the tumorigenicity increase at the 100-μg/kg/day dose is significant, with a P value of <0.021 (χ² test). (B) A549 cells were transfected with the indicated unmodified oligonucleotides (0.4 μM or 0.2 plus 0.2 μM [for the combination]). Eighteen hours after transfection, the cells were transferred to 0.3% agarose containing 0.8 or 0.4 μM (for the combination) the indicated oligonucleotides. The cells were stained with p-iodotetrazolium violet and analyzed with IPLabSpectrum software. The error bars indicate the standard deviation. (C) Morphologies of A549 colonies treated with 0.1 μM rhEGF (right) or untreated (left) and transfected with the indicated oligonucleotides.