Tumor inhibitory activity of anti-ras ribozymes delivered by retroviral gene transfer.

This study reports the successful introduction into tumor cells of a ribozyme directed against an oncogene using a retroviral gene delivery system. A hammerhead ribozyme that selectively targets and cleaves the activated Ha-ras (V12Ras) oncogene was delivered using a retroviral vector and expressed under the control of a transfer RNA promoter in V12Ras-transformed 3T3 murine fibroblast and rat colon epithelial cells. The expression of V12Ras messenger RNA and V12Ras P21 protein was reduced by up to 100% and 75%, respectively, in cells transduced with functional ribozyme. Reductions in V12Ras expression correlated with the stable expression of the functional ribozyme in vivo and decreased tumorigenicity in nude mice. Ribozyme constructs containing identical antisense flanking regions, but a mutant catalytic center, did not attenuate V12Ras expression or decrease the tumorigenicity of transduced tumor cells. These data support the potential therapeutic role of antioncogene ribozymes.