To study drug resistance mechanism of gastrointestinal tumor, doxorubicin resistant cell line of human gastric cancer, we developed BGC-823/DOX, by increasing doses of doxorubicin. We completed chromosomal analysis, double time of cell grow, cell cycle distributions, drug resistance and cross-resistance, and studied reversing drug resistance. The results showed that chromosome of BGC-823/DOX was subtriploid karyotype. Cell morphology, double time, cell cycle distributions were different from its parent cells and 6.35 times more resistant to cytotoxic action of doxorubicin, cross-resistant to VP-16. Verapamil showed modified effect on doxorubicin resistance. BGC-823/DOX with doxorubicin resistance has many characteristics of gastrointestinal tumors.