Purpose: To determine the dose-response associations of sedentary behaviour with cardiovascular diseases (CVD), cancer, and all-cause mortality, and to examine whether the sedentary-associated all-cause mortality risk was affected by appearance of diabetes and hypertension, physical activity, and body mass index (BMI). Design: We carried out a systematic review and meta-analysis to search Medline, SportDiscus, and Web of Science for eligible studies. Settings: Prospective cohort studies that reported sedentary time and CVD, cancer, and mortality incidents. Measurements: Two authors independently extracted data based on predefined criteria. The effect estimates were evaluated by hazard ratios (HRs) with 95% confidences (CIs). Results: Twenty-four studies met the inclusion criteria. Sitting time showed dose-response associations with CVD, cancer, and all-cause mortality, with each 1-hour increment of sitting time daily accounting for HRs 1.04 (95% CIs 1.02-1.07), 1.01 (1.00-1.02), and 1.03 (1.02-1.03), respectively. The link between sitting time and CVD and all-cause mortality was non-linear (p(non-linear) < 0.0001). The relationship between TV viewing and CVD and all-cause mortality was dose-dependent, with HRs 1.07 (1.06-1.09) and 1.04 (1.01-1.06) for per 1-hour increment of TV time every day, respectively. The regression was curved (p(non-linear) < 0.0001). When the analysis was stratified by the percentage of diabetes and hypertension, BMI values, and physical activity levels, we found that higher BMI and a greater percentage of diabetes and hypertension further increased all-cause mortality risk in the most sedentary populations, whereas higher physical activity levels decreased it. Conclusion: Sitting time and TV viewing significantly increased cardiovascular, cancer, and mortality risk; the associations were dose-dependent. More importantly, sedentary behaviour in combination with chronic diseases or high BMI increased all-cause mortality risk whereas physical activity was likely to alleviate the adverse associations.