OBJECTIVE： To investigate the expression difference and its mechanism of miR-27a-3p and HOXB8 protein in cervical cancer tissues of HPV16-positive and HPV16-negative patients. METHODS： A total of 120 patients with cervical cancer in our hospital during Jan. 2012-Jan. 2016 were divided into HPV16-positive group （60 cases） and HPV16-negative group （60 cases） according to HPV16 infection situation. The expression of miR-27a-3p mRNA and HOXB8 mRNA in cervical cancer tissue were detected by RT-qPCR. The expression of HOXB8 protein was detected by Western blotting assay. DNA methylation level of miR-27a-3p promoter region was detected by nested-down methylation specific PCR （nMS-PCR）. Histone methylation level of miR-27a-3p promoter region was detected by chromatin immunoprecipitation PCR （CHIP-PCR）. The expression of miR-27a-3p mRNA and HOXB8 mRNA were detected after transfecting miR-27a-3p mimic and inhibitor into  Human cervical cancer cell line SiHa， respectively. RESULTS： The expression of miR-27a-3p in HPV16-positive group was significantly lower than HPV16-negative group， while HOXB8 mRNA and protein expression， DNA and histone methylation levels of miR-27a-3p promoter region were significantly higher than HPV16-negative group， with statistical significance （P＜0.05 or P＜0.01）. After transfecting miR-27a-3p mimic into SiHa cells， the expression of miR-27a-3p was increased significantly， while that of HOXB8 mRNA was decreased significantly； after miR-27a-3p inhibitor transfection， the expression of miR-27a-3p was decreased significantly， while that of HOXB8 mRNA was increased significantly， with statistical significance （P＜0.01）. CONCLUSIONS： HPV16 may down-regulate the expression of miR-27a-3p through DNA methylation and histone methylation of promoter region， so as to influence the generation and development of cervical cancer. HOXB8 may be the target protein of miR-27a-3p.