Drug resistance testing of acute myeloid leukemia in adults using the MTT assay

We want to evaluate the MTT in vitro assay for newly diagnosed AML in adults, in particular its prognostic significance.

MTT tests were performed according to Pieters et al., Blood 76: 2327, 1990. Up to 18 cytostatic drugs were tested in a wide range of concentrations over 4 days and compared with controls. If necessary, blasts were enriched > 80% by negative selection with dynabeads (Kaspers et al., Br. J. Cancer 70: 1047, 1994). Percent S-phase was measured at begin of assay and after 4 days.

Technical success rate was 80-85% and the assay took about one day of work. IC50 values as a measure for drug resistance were highly reproducible with an interassay CV (range) of 28% (13-47) between days. Median cell viability of controls after 4 days was 80% (44-97);% S-phase at begin of assay was 3% (0.1-15) and after 4 days 9% (1-39). Leukemic blasts from 57 adult AML patients were prospectively tested in vitro at diagnosis. Large interindividual differences in drug resistance were observed (over 1000 fold on average). There was a trend for greater IC50 with higher cytogenetic risk and nonresponders to induction. Other clinical prognostic factors such as patient age, initial WBC, FAB subtype, prior MDS and % S-phase did not show a relationship with the in vitro data. There was a high correlation between the topoisomerase II related drugs regarding in vitro resistance.

In vitro sensitivity tests like the MTT assay may provide a valuable tool for prediction of individual therapy outcome in combination with cytogenetics. However, longer follow-up is required.