To establish a CDDP-resistant cell line from human nasopharyngeal carcinoma and evaluate its biological characteristics.By continuously exposing and gradually increasing dose of cisplatin (CDDP), a resistant nasopharyngeal carcinoma cell line (HNE1/CDDP) was established. Drug sensitivity of this cell line was detected by MTT assay; the alterations of its biological characteristics were determined using light microscopy, cell counting and flow cytometry (FCM); its ability of adhesion, migration and invasion were also evaluated.HNE1/CDDP cell line was developed after 10 months with stable resistance to cisplatin with the resistance index was 5.83. HNE1/CDDP cell exhibited cross-resistance to many other chemotherapeutic agents (carboplatin, oxaliplatin and etoposide, etc). The morphology of HNE1/CDDP changed; doubling time prolonged; and the cell number of S-phase and G2/M-phase decreased while of G0/G1 phase increased compared with parental cells. The ability of adhesion, migration and invasion had no difference between the parental and the resistant cells.HNE1/CDDP cell line shows the typical and stable resistant phenotype and can be used as a research model.