Objective: To develop the method to transfer exogenous genes into oral mucosa epithelial cells with retrovirus. Methods: Oral epithelial cells were isolated from infant oral mucosa with dispase II and trypsin and cultured with DK-SFM in vitro. Then the cells were infected with retrovirus LXSN HPV16E6 and selected with antibiotics G418. Positive clones were passed continuously. PCR and DNA-sequencing were used to detect the E6 sequence. BLAST was performed to contrast the sequence with those in GenBank. Results: Oral epithelial cells grew well in DK-SFM and could be passed eight to nine generations continuously. Before senescence, these cells survived eight to 10 weeks in vitro. HPV16E6 was transferred into epithelial cells with retrovirus. After selection, the survival cells had already been passed eight generations and still grew well. Elecrophoresis indicated there was a specific band of PCR products in a size of 380 bp. DNA sequencing proved the band was equal to E6 sequence. BLAST displayed it was 100% correct to sequences of E6 in GenBank. Conclusion: Oral epithelial cell lines infected with retrovirus LXSN HPV16E6 were established. It was also adequate to transfer other oncogenes into epithelial cells with retrovirus. It was useful to study the roles of oncogenes in the process of cancer development.