Abstract: To meet the requirement of research and development of therapeutic vaccine againsthuman cervical cancer caused by continuous infection of human papillomavirus type 58 (HPV-58),a novel HLA-A*0201 positive human cervical cancer cell model which could expressed HPV-58 oncoprotein E7 was established and characterized.In the first place,the encoding gene of E7 was amplified from the whole-genome of HPV-58 by using PCR,and was further cloned into the vector of pEGFP-N2 to build the recombinant expression plasmid pEGFP-N2-HPV58E7.After being transformed into E.coli DH5α and verified by DNA sequencing,the isolated endotoxin-free plasmid was transformed into human cell line HEK-293T to test its capability of HPV-58 E7 expression by using RT-PCR and immunofluorescence.Furthermore, the E7-negative human cervical cancer cell line C33A which initially expresses human HLA-A*0201 molecular was chose as a host cell to build the target cell model by plasmid transfection,G418 selection and clonal cultivation.Finally,the biological characteristics of cell model,including the correct transcription and expression of HPV-58 E7 and its relatedbiological phenotypes,were evaluated by using RT-PCR,Western-blot,and cell viability assay.As a result,the encoding gene of HPV-58 E7 was correctly cloned and successfully used to build expression vector,leading the effective expression of HPV-58oncoprotein E7 in human cells in a fused form.Moreover,those results of further experimental verification suggested that the target cancer cell model C33A/pEGFP-N2-HPV58E7 was successfully established.The oncoprotein of HPV-58 E7 was not onlycorrectly expressed in this cell model,but it also could be effectively presented by initially expresses HLA-A*0201 molecular for specific recognition of active cytotoxic T lymphocyte.In addition,the viability analysis also revealed that the expressed oncoprotein could effectively stimulate the growth of model cells.In conclusion,the generation of this stably-transfected cell lines would provide a wide range of applications for future research and development of HLA-A*0201 restricted vaccine against HPV-58 associated cervical cancer.