Retroviral recombinants pZip-raf and pZip-myc were constructed to examine the role-of the-c-raf-1 and c-myc proto-oncogene in lung carcinogenesis. Immortalized human bronchial epithelial cells (BEAS-2B) transfected with pZip-raf DNA ar pZip-raf DNA gave rise to undifferentiated carcinomas (raf/myc tumors) when tested in athymic nude mice, whereas c-myc or c-raf transfected cells are nontumorigenic. The raf/myc tumors expressed markers of small cell lung carcinomas i.e., neuron-specific enolase and neurosecretory granules. In addition, BEAS-2B cells transformed with the c-raf and c-myc proto-oncogenes, as well as derived tumor cell lines acquired HLA class II antigen expression. The c-raf-1 gene has been identified as the predominant transforming gene of three radiation-resistant head and neck cancer cell lines in the NIH 3T3 transfection assay (SQ-20B, JSQ-3, SCC- 35). NIH 3T3 cells transformed with SQ-20B DNA also became radiation-resistant, suggesting a correlation between the presence of c-raf sequences and the radiation-resistant phenotype. Inhibition of the c-raf function by introduction of anti-sense raf transcribing plasmids into the SQ-20B cell line reverted not only the tumorigenic phenotype but also reduced the radiation resistance. As a consequence of these experiments, the construction of an inducible promoter system for anti-sense sequences in human cells was undertaken.
