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Retinoic acid-resistant HL-60 cells exclusively contain mutant retinoic
acid receptor-alpha
YP Li, F Said and RE Gallagher
Department of Oncology, Montefiore Medical Center, Albert Einstein Cancer
Center, Bronx, NY 10467.
Sequence analysis of the retinoic acid receptor-alpha (RAR alpha) gene from
a subline of HL-60 cells (RA-res) stably resistant to all-trans retinoic
acid (RA) disclosed a single-base change in codon number 411, the same C to
T transition previously reported in an independently selected HL-60 RA
resistant clone by Robertson et al (Blood 80:1885, 1992). This mutation
eliminates a FokI restriction endonuclease site. Using primers framing this
mutation in exon 9 of the RAR alpha gene, we showed that polymerase chain
reaction products amplified from either mRNA or genomic DNA templates from
the RA-res subline were completely resistant to FokI digestion whereas
those from wild-type (wt) HL-60 cells could be digested to completion. The
lack of a normal allele in the RA-res cells was confirmed by mixing
experiments and hybridization analyses. Southern blot analysis of DNA from
the RA-res and wt cells versus control placental DNA indicated that the RAR
alpha gene is not haploid. The independent isolation of the same RAR alpha
mutation in different laboratories suggests either that the mutation exits
in a small subpopulation in the wt line or that this is a mutational "hot
spot." Furthermore, the results indicate that if a dominant negative mode
of resistance is involved in the RA-res subline, this must involve
interference with the function of heterologous receptor proteins such as
the retinoid X receptors. The lack of any normal RAR alpha in this subline
may facilitate studies of the mode of action of retinoids.
Volume 83,
Issue 11,
pp. 3298-3302,
06/01/1994
Copyright © 1994 by The American Society of Hematology
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