Centrosomal amplification and aneuploidy induced by the antiretroviral drug AZT in hamster and human cells.
The centrosome directs chromosomal migration by a complex process of tubulin-chromatin binding. In this contribution centrosomal abnormalities, including centrosomal amplification, were explored in Chinese hamster ovary (CHO) and normal human mammary epithelial cells (NHMECs) exposed to the antiretroviral drug zidovudine (3'-azido-3'-deoxythymidine, AZT). Centrosomal amplification/fragmentation was observed in both cell types and kinetochore positive micronuclei were found in AZT-exposed CHO cells in correlation with dose. Normal human mammary epithelial cell (NMHEC) strain M99005, previously identified as a strain that incorporates high levels of AZT into DNA (high incorporator, HI), showed greater centrosomal amplification when compared with a second strain, NHMEC M98040, which did not incorporate AZT into DNA (low incorporator, LI). Additionally, an abnormal tubulin distribution was observed in AZT-exposed HI cells bearing multiple centrosomes. Immunofluorescent staining of human cells with Aurora A, a kinase involved in the maturation of the centrosome, confirmed the induction of centrosomal amplification and revealed multipolar mitotic figures. Flow cytometric studies revealed that cells bearing abnormal numbers of centrosomes and abnormal tubulin distribution had similar S-phase percentages suggesting that cells bearing unbalanced chromosomal segregation could divide. Therefore, AZT induces genomic instability and clastogenicity as well as alterations in proteins involved in centrosomal activation, all of which may contribute to the carcinogenic properties of this compound.
Laboratory of Cancer Biology and Genetics, CCR, NCI, NIH, Bethesda, MD 20892, USA.
Jennifer P
JP
Borojerdi
Jessica
J
Ming
Catherine
C
Cooch
Yvona
Y
Ward
Cristina
C
Semino-Mora
Mia
M
Yu
Hannan M
HM
Braun
Barbara J
BJ
Taylor
Miriam C
MC
Poirier
Ofelia A
OA
Olivero
D024422:Aneugens / Q000493:pharmacokinetics / Q000633:toxicity*; D000782:Aneuploidy; D000818:Animals; D064127:Aurora Kinases; D001940:Breast / Q000166:cytology / Q000187:drug effects / Q000378:metabolism; D016466:CHO Cells; D002453:Cell Cycle / Q000187:drug effects; D002460:Cell Line; D018385:Centrosome / Q000187:drug effects* / Q000378:metabolism / Q000648:ultrastructure; D006224:Cricetinae; D003412:Cricetulus; D018736:DNA Adducts / Q000378:metabolism; D004847:Epithelial Cells / Q000187:drug effects / Q000378:metabolism / Q000648:ultrastructure; D006801:Humans; D048629:Micronuclei, Chromosome-Defective / Q000139:chemically induced; D046529:Microscopy, Electron, Transmission; D017346:Protein Serine-Threonine Kinases / Q000378:metabolism; D014404:Tubulin / Q000378:metabolism; D015215:Zidovudine / Q000493:pharmacokinetics / Q000633:toxicity*
D024422:Aneugens; D018736:DNA Adducts; D014404:Tubulin; D015215:Zidovudine; D064127:Aurora Kinases; D017346:Protein Serine-Threonine Kinases