A search for nuclear RNA lost in azo dye-induced transplantable hepatoma

Studies were conducted to characterize the nuclear RNA (nRNA) species that were present in rat liver but absent in the hepatoma. Nuclear RNA was compared between Donryu rat liver and AH136B hepatoma, an azo dye-induced transplantable cell line, by DNA-RNA competitive hybridization. The hepatoma lacked 13-14% of nRNA according to measurements of radioactivity of the hybridized 32P-labeled liver nRNA, and this loss was shown to be due to the failure to transcribe such RNA rather than to the deletion of the relevant DNA in the genome. Characterization of the lost RNA was first attempted by fractionating liver nRNA by density gradient sedimentation and polyacrylamide gel electrophoresis. A comparison of the additive effects of the fractionated RNA's in the competitive hybridization indicated that the pertinent RNA was present in the large RNA molecules (greater than 14S), not in the low molecular weight RNA's. Then poly(A) nRNA was found to show a strong additive effect in the competitive hybridization while nucleolar RNA showed little additive effect, indicating that the pertinent RNA was present in the heterogeneous nRNA, not in the ribosomal precursor. Further characterization was done by fractionating DNA with regard to the repetition in the genome. A comparison of the competitive hybridizations on the fractionated DNA's showed that the loss occurred mostly in RNA transcribed from highly repetitive DNA. In conclusion, the RNA species lost in the hepatoma were components of heterogeneous nRNA transcribed from highly repetitive DNA.