The feasibility of treating solid tumours with differentiation therapy using antineoplastic drugs is currently being investigated, but the emergence of multidrug resistance remains the major limitation to this therapeutic approach. A rhabdomyosarcoma cell line resistant to actinomycin D (RD-DAC) has been used as an in vitro model to investigate, with light and electron microscopy, the degree of differentiation in multidrug-resistant cells. The parental cell line (RD), derived from a human embryonic-type rhabdomyosarcoma, is undifferentiated, with no evidence of specific ultrastructural markers. Examination of resistant cells by transmission electron microscopy revealed myofilaments arranged parallel to the long axis of the cell, which was considered clear evidence of myogenic differentiation. These observations suggest that actinomycin D, the drug of choice in the treatment of rhabdomyosarcoma, induces differentiation in the cell line RD. It is postulated that multidrug resistance can interfere with cellular differentiation.