INSERM U268; Hôpital Paul Brousse; 94800, Villejuif, France

^aAuthor for correspondence: INSERM U268, Hôpital Paul-Brousse, 14 avenue Paul Vaillant Couturier, 94807 Villejuif cedex, France. tel: (33) 01 45 59 53 09; fax: (33) 01 45 59 53 29; e-mail: jrl@infobiogen.fr

Edited by A. Columbano

Friend murine erythroleukaemia (F-MEL) cells are a useful model for studying the processes that regulate erythroid differentiation since exposure of these cells to chemical inducers (DMSO or HMBA) results in commitment to terminal cell division and synthesis of haemoglobin. This study examined the relationship between differentiation and apoptosis in DMSO sensitive and resistant F-MEL cells. Clear apoptosis was not observed in DMSO-treated sensitive F-MEL (strain 745A) cells during the induction of differentiation. In contrast, DMSO-induced 745A cells exhibited delayed apoptosis compared to uninduced cells. Since the Bcl-2 family members play a major role in the control of apoptosis and/or differentiation, we determined their expression before and after DMSO or HMBA treatment. Neither untreated nor chemically-induced 745A cells expressed the Bcl-2 protein. The levels of Bax and Bad proteins remained relatively constant during DMSO-induced differentiation. DMSO or HMBA treatment of 745A cells induced a marked increase of Bcl-X_L expression during the late phase of differentiation which persisted even when the cells began to die. This upregulation of Bcl-X_L was independent of cell density but was correlated with cell arrest in G0/G1. DMSO treatment induced a similar delay of apoptosis and enhancement of Bcl-X_L expression in F-MEL (strain TFP10) cells which fail to synthesize haemoglobin in the presence of DMSO. Dexamethasone, which blocks DMSO-induced differentiation of F-MEL cells, prevented the induction of Bcl-X_L. Inhibitors such as imidazole or succinylacetone, which inhibit haemoglobin synthesis but not commitment to terminal cell division, did not suppress Bcl-X_L induction in DMSO-induced cells. Taken together, these results indicate that DMSO treatment of F-MEL cells induces a marked increase in Bcl-X_L expression suggesting a role for this anti-apoptotic protein in the process of erythroid differentiation in F-MEL cells. Moreover, induction of Bcl-X_L during this process seems to be associated with loss of proliferative capacity rather than with haemoglobin synthesis.

apoptosis; Bcl-X_L; erythroid differentiation; Friend erythroleukaemia; haemoglobin synthesis