Altered epidermal growth factor signal transduction in activated Ha-ras-transformed human keratinocytes

Epidermal growth factor (EGF) can stimulate proliferation and 92 kDa gelatinase/matrix metalloproteinase (MMP-9) expression. The induction of MMP-9 is not only pathologically significant for invasion and metastasis, but also serves as a semiquantitative measure of EGF signal transduction. In order to examine the role of mutated ras p21 in EGF signal transduction, an activated Ha-ras-transformed human keratinocyte cell line was developed and characterized. Overexpression of the mutated Ha-ras p21 in these cells was demonstrated. Our results showed that EGF induced 92 kDa MMP-9 secretion was doubled in the ras-transformed keratinocytes in comparison to the parent cells. The karyotype, the expression of EGF receptor (EGFR) and transforming growth factor (TGF) alpha at the mRNA level remained unchanged. These results suggest that the presence of high levels of mutated ras p21 may be responsible for the aberrant EGF signal transduction and contributes to transformation. In addition, a reduction of TGF beta expression at mRNA level by 70% was found in the activated Ha-ras-transformed keratinocytes when compared to the parent cells.