In vitro study of the combination gemcitabine + fludarabine on freshly isolated chronic lymphocytic leukemia cells.
OBJECTIVE
Fludarabine has shown a definite clinical activity in B-cell chronic lymphocytic leukemia (CLL). If the effects of this drug could be potentiated, it could be useful in order to obtain complete remissions. In this study we evaluated the effects of the combination of fludarabine and gemcitabine, a deoxycytidine analog that has shown both in vitro and in vivo activity against a variety of solid tumors.


METHODS
CLL cells from 10 patients were cultured in vitro in the presence of fludarabine (0.5-1,000 microg/mL) and gemcitabine (0.1-5,000 microg/mL), both alone and in different combinations. Cytotoxic activity was tested by the XTT colorimetric assay. Furthermore we evaluated BCL-2 protein expression and, subsequently, the induction of apoptosis at baseline and after exposing cells to different concentrations of fludarabine and gemcitabine.


RESULTS
The IC(50) of fludarabine and gemcitabine on CLL cells was 550 and 1,100 microg/mL, respectively, in our series of samples; the cytotoxicity of either drug was not influenced by the percentage of BCL-2 positive cells in the same sample. The addition of gemcitabine increased fludarabine-induced cytotoxicity; however, isobologram analysis of the data showed synergism only when lower doses of gemcitabine were combined to fludarabine. Induction of apoptosis reflected this pattern of activity.


CONCLUSIONS
Gemcitabine was able to increase the activity of fludarabine only when low doses of the former were employed. As both compounds incorporate into DNA blocking chain elongation, our results could be explained by the drugs interfering at that level. The possibility of potentiating the effects of fludarabine with low doses of gemcitabine renders this combination promising in view of an in vivo use.
Tosi
P
P
Istituto di Ematologia ed Oncologia Medica "L. e A. Seràgnoli", Policlinico S. Orsola, via Massarenti, 9, 40138 Bologna, Italy ptosi@med.unibo.it
Pellacani
A
A
Zinzani
P L
PL
Magagnoli
M
M
Visani
G
G
Tura
S
S
D000964:Antimetabolites, Antineoplastic / Q000494:pharmacology*; D017209:Apoptosis / Q000187:drug effects; D004273:DNA, Neoplasm / Q000378:metabolism; D003841:Deoxycytidine / Q000031:analogs & derivatives* / Q000494:pharmacology; D004357:Drug Synergism; D015973:Gene Expression Regulation, Leukemic / Q000187:drug effects; D019254:Genes, bcl-2; D006801:Humans; D015451:Leukemia, Lymphocytic, Chronic, B-Cell / Q000473:pathology*; D009363:Neoplasm Proteins / Q000096:biosynthesis / Q000235:genetics; D014411:Neoplastic Stem Cells / Q000187:drug effects*; D019253:Proto-Oncogene Proteins c-bcl-2 / Q000096:biosynthesis; D014740:Vidarabine / Q000031:analogs & derivatives* / Q000494:pharmacology; D000093542:Gemcitabine
D000964:Antimetabolites, Antineoplastic; D004273:DNA, Neoplasm; D009363:Neoplasm Proteins; D019253:Proto-Oncogene Proteins c-bcl-2; D003841:Deoxycytidine; D014740:Vidarabine; C024352:fludarabine; D000093542:Gemcitabine