Background: It is well established that a healthy lifestyle is of benefit in the prevention of diseases such as cancer and promotion of well-being. Therefore, the objective of this study was to determine lifestyle practice and associated factors among university students in Malaysia. Materials and Methods: A cross sectional study was conducted over six months from November 2011 until May 2012 among the students from the Management and Science University. This study was approved by its ethical committee, the students being explained the objective and invited to participate. A consent form was signed by all study participants. Questionnaire was distributed randomly to the students of the five faculties through their lecturers in different faculty. For univariate analysis t-test and ANOVA test were performed. Multiple linear regression used for multivariate analysis using SPSS 13.0. Results: A total number of 1100 students participated with a mean age of $22.1{pm}2.21$ (SD) years. The majority were 22 years or younger (56.3%), female (54%), Malay (61.5%), single (92.3%), with family monthly income ${geq}5000$ Ringgit Malaysia (41.2%). Regarding lifestyle, about were 31.6% smokers, 75.6% never drank alcohol and 53.7% never exercised. Multivariate analysis showed that age, sex, race, parent marital status, participant marital status, type of faculty, living status, smoking status, exercise, residency, brushing teeth, fiber intake and avoid fatty food significantly influenced the practice of drinking alcohol among university students (p=0.006, p=0.042, p<0.001, p=0.003, p=0.002, p<0.001, p<0.001, p<0.001, p<0.001, p<0.001, p<0.001, p=0.003, p<0.001; respectively). It similarly showed that sex, race, parent marital status, participant marital status, monthly family income, exercise, residency, brushing teeth and fiber intake significantly influenced the practice of sun protection (p<0.001, p<0.001, p<0.001, p<0.001, p<0.001, p=0.017, p<0.001, p<0.001, p<0.001; respectively) and that age, sex, parent marital status, participant marital status, type of faculty, living status, exercise, taking non-prescribed medication, brushing the teeth, coffee consumption and fiber intake were significantly influenced the practice of fruits consumption (p=0.008, p<0.001, p<0.001, p<0.001, p<0.001, p<0.001, p<0.001, p<0.001, p=0.002, p<0.001, P<0.001; respectively). Conclusions: This study showed a poor practice of healthy lifestyle among university students. Therefore universities should emphasize a healthy lifestyle in all faculties as a required subject. Socio-demographic characteristics significantly influenced practice and thus should be considered when planning preventive measures among university students. Frequent campaigns and educational seminars are to be encouraged.