OBJECTIVE To investigate the effect of complex amino acid imbalance on the growth of tumor in tumor-bearing (TB) rats. METHODS Sprague-Dawley (SD) rats underwent jejunostomy for nutritional support. A suspension of Walker-256 carcinosarcoma cells was subcutaneously inoculated. TB rats were randomly divided into groups A, B, C and D according to the formula of amino acids in enteral nutritional solutions, respectively. TB rats received jejunal feedings supplemented with balanced amino acids (group A), methionine-depleted amino acids (group B), valine-depleted amino acids (group C) and methionine- and valine-depleted complex amino acid imbalance (group D) for 10 days. Tumor volume, inhibitory rates of tumor, cell cycle and life span of TB rats were investigated. RESULTS The G0/G1 ratio of tumor cells in group D (80.5 +/- 9.0)% was higher than that in groups A, B and C which was 67.0 +/- 5.1%, 78.9 +/- 8.5%, 69.2 +/- 6.2%, respectively (P<0.05). The ratio of S/G2M and PI in group D were lower than those in groups A, B and C. The inhibitory rate of tumor in groups B, C and D was 37.2%, 33.3% and 43.9%, respectively (P<0.05). The life span of TB rats in group D was significantly longer than that in groups B, C, and A. CONCLUSIONS Methionine/valine-depleted amino acid imbalance can inhibit tumor growth. Complex amino acids of methionine and valine depleted imbalance have stronger inhibitory effects on tumor growth.