Selective inhibition of human ribosomal gene transcription by the morpholinyl anthracyclines cyanomorpholinyl- and morpholinyldoxorubicin.
Upon incubation of cultured mammalian cells with the new anthracycline analogues cyanomorpholinyldoxorubicin and morpholinyldoxorubicin, nucleoli irreversibly segregate into their substructures which form individual portions of the nucleolar mass and characteristic electron-dense components adjacent to the nucleolonema; these changes in nucleolar ultrastructure are similar to those produced by actinomycin D (AMD). In the present study we have examined the effects of anthracycline analogues on RNA synthesis, localization of RNA polymerase I in situ, and activity of RNA polymerases in vitro, and compared these effects with those of the parent compound doxorubicin (DOX) and AMD. The results show that, following treatment with cyanomorpholinyldoxorubicin, morpholinyldoxorubicin, and AMD, but not DOX, RNA polymerase I-containing transcription complexes were reduced, reflecting the transcriptional activity of the rRNA genes. The residual RNA polymerase-containing entities were redistributed into cap-like aggregates at the nucleolar periphery. Within 30 min of exposure to cyanomorpholinyldoxorubicin, morpholinyldoxorubicin, and AMD, but not DOX, a 75-90% inhibition of RNA polymerase I activity in situ and in vitro was observed. At this early time there was no significant inhibition of nucleoplasmic RNA labeling in situ or RNA polymerases II and III activities in vitro. At later times following reincubation in drug-free medium, inhibition of all three polymerases was observed. Impairment of RNA synthesis appeared to result from drug interaction with the DNA template rather than an interaction with RNA polymerase I itself. We conclude that the morpholinyl derivatives of DOX are preferential inhibitors of ribosomal gene transcription and that they may have a mechanism of action similar to that of AMD on rRNA synthesis.
Wassermann
K
K
Department of Medical Oncology, University of Texas M. D. Anderson Hospital and Tumor Institute, Houston 77030.
Newman
R A
RA
Davis
F M
FM
Mullins
T D
TD
Rose
K M
KM
D003609:Dactinomycin / Q000494:pharmacology; D004317:Doxorubicin / Q000031:analogs & derivatives* / Q000494:pharmacology; D005455:Fluorescent Antibody Technique; D006367:HeLa Cells / Q000201:enzymology; D006801:Humans; D008854:Microscopy, Electron; D012318:RNA Polymerase I / Q000378:metabolism; D012270:Ribosomes / Q000187:drug effects*; D014158:Transcription, Genetic / Q000187:drug effects*
D003609:Dactinomycin; D004317:Doxorubicin; C048396:NSC 354646; C038555:3'-deamino-3'-(3-cyano-4-morpholinyl)doxorubicin; D012318:RNA Polymerase I