ACUTE PROMYELOCYTIC LEUKEMIA-CELLS RESISTANT TO RETINOIC ACID SHOW FURTHER PERTURBATION OF THE RAR-ALPHA SIGNAL-TRANSDUCTION SYSTEM

Acute promyelocytic leukaemia (APL) cell lines resistant to all-trans retinoic acid (ATRA) have been previously derived from the NB4 cell line, and characterized as having lost the expression of the intact pml/RAR alpha fusion protein. To confirm the association between ATRA-resistance and alteration in the fusion protein at the clonal level, 16 clones were generated from ATRA-resistant APL cell lines. All clones show immunological (HLA class I and II, CD11b and c, CD13 and 33), molecular and growth features similar to the parental cell lines. To investigate whether the irradiation protocol used tb generate the previously reported retinoic acid-resistant NB4.306 cell line induced additional alterations that could render these cells able to escape the anti-proliferative effect of retinoic acid (ATRA), an additional ATRA-resistant APL cell line, [NB4.007/6], was generated, under the selective pressure of ATRA, from the NB4 cell line without previous radiation. This cell line shows resistance to the antiproliferative and differentiating action of ATRA. The NB4.007/6 cell line contains the t(15;17) chromosome translocation, shows the usual pml/RAR alpha hybrid DNA but expresses no detectable amount of the usual pml/RAR alpha protein in Western blot analysis, similarly to the NB4.306 cell line. Finally, the relative resistance to ATRA of NB4.306 and NB4.007/6 was evaluated by comparing the phenotypic (CD11b) changes induced by ATRA in these two lines with those induced in the parental, ATRA-sensitive, NB4 cell line. It is estimated that NB4.306 and NB4.007/6 are about 300 and 70 times less sensitive to ATRA than the original NB4 cell line.