Abstract: Objective：Constructing retroviral vector of small hairpin RNA （shRNA） targeting human transducer and activator of transcription 3（STAT3），establishing stable infected HPV16 positive cervical cancer cell lines，to further explore the role of STAT3 in the development of HPV16 positive cervical cancer. Methods：Design and synthetise three siRNA sequences targeting STAT3，then select one with best gene interference effect，and construct STAT3-shRNA vector，pSUPERretro-puro-STAT3. The recombinant plasmid was transfected into packaging cell line 293FT and then infected into HPV16 positive cervical cancer cell line SiHa and CaSki cells. Cells were selected with puromycin for 2 weeks，passaged around every 3 days，and named with SiHa/STAT3-sh and CaSki/STAT3-sh. Expression of STAT3 was analyzed by quantitative reverse transcription polymerase chain reaction （qRT-PCR） and western blotting. Results：The knockdown of the expression of STAT3 mRNA was over 99%（P＜0.01），that of the expression of STAT3 total protein was over 70% （P＜0.01） and that of the expression of STAT3 phosphorylated protein was over 78% （P＜0.01） in both SiHa/STAT3-sh and CaSki/STAT3-sh cells． Conclusions：HPV16 positive cervical cancer cell lines with stable silence of STAT3 were successfully established.

Key words: Uterine cervical neoplasms, Carcinoma, DNA-binding proteins, Human papillomavirus 16, RNA interference, Retroviridae