Objectives: Breast cancer is the leading cause of death among Saudi females. Knowledge and awareness regarding breast cancer would improve patients’ quality of life through early detection of the disease. This study aimed to measure female university students’ knowledge and practice regarding breast cancer. Subjects and Methods: A cross‑sectional research design was adopted in this study. A prevalidated questionnaire was developed and distributed to 900 female students using nonprobability convenience sampling technique. A total of 826 valid questionnaires were successfully obtained. Female students were met in the public areas of the females’ campus. All data were analyzed using SPSS version 22 at alpha value of 0.05. Results: Majority of respondents showed low level of knowledge regarding breast cancer. The Internet was the main source of knowledge 53.5%. Moderate knowledge regarding breast cancer symptoms were observed by more than 50% of respondents. Knowledge regarding risk factors was low by majority of respondents >50%. In addition, only 28% of respondents practice breast self‑examination (BSE), whereas main barrier for not practicing BSE was the lack of knowledge on how to perform BSE 57.5%. Conclusion: Students’ knowledge and practice toward breast cancer was inadequate. There is a need for national campaigns for the public and university female students to improve their knowledge regarding breast cancer and encourage them for practicing BSE.