Bovine arterial endothelial cells were stably transfected with the human wild-type (wt) HSP27 or a mutant gene (mu) encoding a nonphosphorylatable form of the protein. At early passage both cultural and cellular morphology were similar, although the vacuole content in wtHSP27 was much higher than muHSP27 cells. As the cultures aged, wtHSP27 cells became large, polymorphic, highly vacuolated, and reached senescence before muHSP27 transfected cultures, which remained small and polygonal with few detectable vacuoles. Vector control cells showed an intermediate phenotype. Tritiated thymidine incorporation studies were performed with multiple wtHSP27 and muHSP27 clones and the results compared with 11 vector control clones. The results showed an average increase in growth rate for the wtHSP27 cells of 3.0 +/- 0.6 times. The growth rate of eight muHSP27 clones showed a slight decrease. Estradiol treatment of endothelial cells resulted in an increase in both bovine and human HSP27, with peak expression at 100 nM. Treatment of the vector-transfected cells with 100 nM estradiol resulted in a 1.44 +/- 0.18 fold increase in growth rate, which was blocked by expression of muHSP27. These data demonstrate a role for HSP27 in controlling the growth rate of endothelial cells in an estrogen-responsive manner.