Recently, we demonstrated that biomechanical stress induces apoptosis of vascular smooth muscle cells (SMCs) (Mayr et al., FASEB J. 2000; 15:261.270). In this article we investigated the molecular mechanisms of mechanical stress‐induced apoptosis. When SMCs were subjected to cyclic strain, tumor‐suppressor p53 was activated as evidenced by gel mobility shift assays and Western blot analyses. p53 activation was largely attenuated if SMCs were pretreated with SB202190, a specific p38MAPK inhibitor, or were stably transfected with dominant negative rac, an upstream signal transducer of p38MAPK pathways. Kinase assays provided direct evidence that p38MAPKs phosphorylated p53 within 30 min of cyclic strain. Additionally, mechanical stress resulted in oxidative DNA damage as detected by the presence of 8‐oxoguanine. Treatment with the antioxidant U‐74389G abrogated p53 activation. p53 activation was followed by expression and mitochondrial translocation of the proapoptotic protein Bax. Likewise, mechanical stress resulted in up‐regulation of anti‐apoptotic Bcl‐2 proteins, including Bcl‐2 and Bcl‐xL. However, a marked loss of mitochondrial membrane potential occurred in wild‐type, but not in p53−/−, SMCs. The latter lost their ability to express Bax and showed no apoptosis in response to cyclic strain. Taken together, our data provide the first evidence that SMC apoptosis induced by mechanical stress is p53‐dependent.