Chemotherapeutic drugs sensitize pre-B ALL cells for CD95- and cytotoxic T-lymphocyte-mediated apoptosis

Abstract

The CD95 (APO-1/Fas) system plays an important role in lymphocyte homeostasis and contributes to anticancer drug-induced apoptosis in some tumor cells. Most childhood B-lineage ALL cells are constitutively resistant towards CD95-induced death. We report here that chemotherapeutic drugs, such as doxorubicin, cytarabine, methotrexate and 6-mercaptopurine, sensitize CD95-resistant pre-B-ALL cell lines for CD95- and lymphokine-activated killer (LAK)-induced cell death. Enhanced susceptibility in drug-treated cells was found to be associated with increased expression of CD95 mRNA and surface expression of CD95 protein, as well as loss of Bcl-x_L protein and disturbance of mitochondrial function. Low level activation of caspases and cleavage of poly(ADP-ribose) polymerase following CD95 triggering was strongly increased in drug pre-treated cells. Furthermore, drug pre-treated cells could be rescued from CD95-mediated apoptosis by blocking the CD95-signaling pathway with a FADD-dominant negative expression construct. Our data suggest that chemotherapeutic drugs may sensitize pre-B ALL cells by at least two mechanisms: (1) by increasing CD95 expression; and (2) by lowering the threshold for apoptotic signals. These findings may explain the effectiveness of low-dose chemotherapy and indicate an active role for key molecules of apoptosis and the immune system during chemotherapy of leukemia.