Combination of Tumor Necrosis Factor Alpha and Interferon Alpha Induces Apoptotic Cell Death through a c-myc–Dependent Pathway in p53 Mutant H226br Non–Small-Cell Lung Cancer Cell Line

We investigated the role of wild-type p53 and c-myc activity in apoptosis induced by a combination of natural human tumor necrosis factor alpha (TNF-α) and natural human interferon alpha (IFN-α). Studies were performed with two human non–small-cell lung cancer cell lines, H226b, which has wild-type p53, and H226br, which has a mutant p53. The combination of IFN-α and TNF-α significantly inhibited cell growth and induced apoptotic cell death of both H226b and H226br, compared with IFN-α or TNF-α alone. Treatment with one or both cytokines did not affect the expression level of p53 in both cell lines. These results suggest that the combination of IFN-α/TNF-α induces apoptotic cell death through a p53- independent pathway. The c-myc oncogene is known to be involved in apoptosis induced by TNF. Antisense c-myc oligonucleotides have been reported to modulate cell growth or apoptosis in several cell lines. Antisense oligodeoxynucleotides were added to the culture of H226br cells before the addition of IFN-α/TNF-α. Antisense c-myc inhibited IFN-α/TNF-α cytotoxicity and apoptotic cell death. In conclusion, this study provides support for the speculation that TNF-α/IFN-α induce apoptosis through a c-myc–dependent pathway rather than a p53-dependent pathway.