Tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) is a multi-functional cytokine, which is involved in the pathophysiological processes of cardiovascular and metabolic diseases. Previously, we demonstrated that TRAIL stimulated lipid uptake and foam cell formation in macrophages in vitro. Several clinical studies have suggested that the serum concentration of TRAIL may be increased in humans with elevated blood cholesterol; however, the current data appear to be inconclusive in this regard. In the present study, we examined the relationships between the serum TRAIL concentration and cholesterol levels in 352 generally healthy subjects undergoing the routine annual health check. We showed that there were significant correlations between TRAIL concentration and levels of total and low-density lipoprotein cholesterols. The level of TRAIL was significantly elevated in subjects with hypercholesterolemia, although this relationship might be also associated with changes of other metabolic factors. Moreover, we showed that the level of blood cholesterol was significantly higher in subjects in the upper quartile of serum TRAIL. In conclusion, our data demonstrate that the serum TRAIL concentration is elevated in people with hypercholesterolemia.