In our previous study, bradykinin (BK) exerts its mitogenic effect through Ras/Raf/MEK/MAPK pathway in vascular smooth muscle cells (VSMCs). In addition to this pathway, the non‐receptor tyrosine kinases (Src), EGF receptor (EGFR), and phosphatidylinositol 3‐kinase (PI3‐K) have been implicated in linking a variety of G‐protein coupled receptors to MAPK cascades. Here, we investigated whether these different mechanisms participating in BK‐induced activation of p42/p44 MAPK and cell proliferation in VSMCs. We initially observed that BK‐ and EGF‐dependent activation of Src, EGFR, Akt, and p42/p44 MAPK and [³H]thymidine incorporation were mediated by Src and EGFR, because the Src inhibitor PP1 and EGFR kinase inhibitor AG1478 abrogated BK‐ and EGF‐dependent effects. Inhibition of PI3‐K by LY294002 attenuated BK‐induced Akt and p42/p44 MAPK phosphorylation and [³H]thymidine incorporation, but had no effect on EGFR phosphorylation, suggesting that EGFR may be an upstream component of PI3‐K/Akt and MAPK in these responses. This hypothesis was supported by the tranfection with dominant negative plasmids of p85 and Akt which significantly attenuated BK‐induced Akt and p42/p44 MAPK phosphorylation. Pretreatment with U0126 (a MEK1/2 inhibitor) attenuated the p42/p44 MAPK phosphorylation and [³H]thymidine incorporation stimulated by BK, but had no effect on Akt activation. Moreover, BK‐induced transactivation of EGFR and cell proliferation was blocked by matrix metalloproteinase inhibitor GM6001. These results suggest that, in VSMCs, the mechanism of BK‐stimulated activation of p42/p44 MAPK and cell proliferation was mediated, at least in part, through activation of Src family kinases, EGFR transactivation, and PI3‐K/Akt. Copyright © 2004 Wiley‐Liss, Inc.