The expansion of chemotherapy raised concerns about the health and safety of hospital personnel. Very little is known about the conditions of handling of chemotherapeutic agents by healthcare workers in Greece and possible adverse effects related to their safety practices, as well as the safety policies adopted by the Greek hospitals. A self-evaluation questionnaire was completed by 353 healthcare workers involved with the use of chemotherapeutic drugs in 24 Greek hospitals and the answers were statistically analysed. The majority of the healthcare workers are aware of the dangers of their work, although they had received limited training and medical surveillance. A significant percentage of them does not use personal protective equipment or use it inadequately. The safety design of their workplace is rather poor. Different health problems have been experienced, deriving from the respiratory, central nervous system, reproductive, gastrointestinal and musculoskeletal system. The improvement of safety training and procedures as well as medical surveillance seems to be a vital priority of hospital administration in Greece, in order to comply with the European guidelines and for the prevention of occupational diseases and environmental pollution.