Background Testicular cancer (TC) is the most common malignancy and the third leading cause of cancer deaths in male adolescents and young adults. Early detection of testicular cancer can be achieved by testicular self-examination (TSE). To date, however, no population-based studies examined TC knowledge and practice of TSE among Polish men. The objective of the study was to assess the level of TC knowledge and awareness as well as the altitude and practice towards TSE among Polish male high school and medical students.Methods A questionnaire survey was conducted in Poland with a representative sample of 1077 male students: 335 from high school and 742 medical students. An original questionnaire was designed as a multiple-choice question type and evaluated the participants’ knowledge and awareness of TC, opinion and attitudes towards performing TSE.Results The knowledge about TC epidemiology and the awareness of risk factors responsible for the TC was low. The knowledge of the signs and symptoms of TC was significantly higher among the medical students, compared to the high school students. The level of education was associated with the awareness of methods for early detection and symptoms of TC. A satisfactory level of awareness of the TSE practice was exhibited only by the medical students. The main reason for not performing self-examination was the lack of knowledge and practical skills.Conclusions The study reflects the deficiencies in health education among high school and medical students. The deficits of knowledge of TC in young men should motivate the education policy makers in Poland to implement education in the field of TC issues more widely in high schools. Moreover, cancer prevention modules and/or teaching methods should be improved in medical schools.