Expression of Bcl-2-related genes in normal and AML progenitors: changes induced by chemotherapy and retinoic acid

The expression of Bcl-2 family members was examined in normal and leukemic hematopoietic cells. Immature hematopoietic progenitor cells (CD34(-)/33(-)/13(-)) did not express Bcl-2 but Bcl-X-L, the majority of CD34 cells expressed Bcl-2, Bcl-X-L and BAD, and normal promyelocytes (CD34(-)/33(+)) lacked expression of both Bcl-2 and Bcl-X-L, while leukemic CD34(+) progenitors and promyelocytes expressed these anti-apoptotic proteins. In AML, Bcl-2 expression was higher on CD34(+) than on all AML cells, however, expression of Bcl-2 or Bcl-X-L did not predict achievement of complete remission, Surprisingly, low Bcl-2 content was associated with poor survival in a group of patients with poor prognosis cytogenetics. The anti-apoptotic BAD protein was found to be expressed in AML, but was phosphorylated in 41/42 samples. Phosphorylation was found at both sites, Ser 112 and Ser 136, During induction chemotherapy, Bcl-2 levels of CD34 cells increased significantly. In the context of evidence for small numbers of leukemic CD34(+) cells expressing very high levels of Bcl-2 prior to therapy, this finding is interpreted as a survival advantage of Bcl-2 overexpressing progenitors and rapid elimination of cells with low Bcl-2 Bcl-2 and Bcl-X-L, were both expressed in minimal residual disease cells. Downregulation of Bcl-2 mRNA and protein was observed by ATRA and the combination of Ara-C, followed by ATRA, resulted in markedly increased cytotoxicity in HL-60 cells, as compared to Ara-C alone or ATRA followed by Ara-C, Implications of these findings for the development of new therapeutic strategies for AML are discussed.