Objective: To evaluate the antitumor activity of recombinant SEA for therapy of B16 melanoma established in C57BL/6 mice. Methods: C57BL/6 mice with melanoma were treated with the purified rSEA. The tumors were isolated and weighted. Results: Tumor growth was apparently inhibited by rSEA at high, middle, and low doses intraperitoneally, whose inhibition ratio were 79.3%, 75.6 % and 73.8% respectively. rSEA treatment in situ could inhibit tumor growth more effectively(90.6%). Further study showed that numerous CD8+ and CD4+ T cell were infiltrated in tumor tissues, which were consistent with tumor growth inhibition induced by rSEA. Conclusions: rSEA could inhibit tumor growth effectively, especially the treatment in situ. This study paves the way for tumor immunotherapy with targeted SEA.