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Objective To investigate the relationships of T→C mutation of 3264 site in tRNAleu region of mitochondrial DNA (mtDNA) with tumorigenesis and p53 mutation. Methods The whole DNA was extracted from carcinoma and adjacent tissues of 28 patients with liver carcinoma and 35 patients with colon cancer. Then tRNAleu region of mtDNA and p53 gene were amplified by PCR and sequenced. Results No T→C mutation was found in 3264 site, but 16 cases of mutations in the other sites were discovered in the carcinoma tissues. No T→C mutation was found in 3264 site, but 3 cases of mutations were discovered in the adjacent tissues .Seventeen cases of p53 gene mutation were found in the carcinoma tissues; 9 cases of p53 mutation were found in patients with colon carcinoma , including 6 cases of tRNAleu mutation and 3 cases of non-tRNAleu mutation (P<0.05); and 8 cases of p53 mutation were found in patients with liver cancer , including 5 cases of tRNAleu mutation and 3 cases of non-tRNAleu mutation (P<0.05). Conclusions The T→C mutation in 3264 site of tRNAleu region can′t be considered to have relationship with tumorigenesis. The mutations in other sites of tRNAleu region are due to p53 mutation, so the gene loses its protection, and thus the mutation occurs. [ABSTRACT FROM AUTHOR]