〔Abstract〕 Objective To investigate the role and mechanism of circular RNA containing pleckstrin homology domain of Pleckstrin homology domain family M member 3(circRNA＿PLEKHM3) in regulating epithelial-mesenchymal transition(EMT) behavior in cervical cancer cells through the miR-320 and KLF4. Methods The expression levels of circRNA＿PLEKHM3 in cervical cancer cells Hela and CaSki were detected by real-time quantitative PCR(qRT-PCR). RNA fluorescence in situ hybridization was used to determine the localization of circRNA＿PLEKHM3 in human cervical cancer epithelial cells CaSki. Dual luciferase reporter gene experiments were conducted to investigate the targeting relationship betweencircRNA＿PLEKHM3andmiR-320, as well as the targeting relationship betweenmiR-320andKLF4. CaSki cells were overexpressed withcircRNA＿PLEKHM3. Additionally, three groups were set up: overexpression ofmiR-320on the basis ofcircRNA＿PLEKHM3overexpression, silencing ofKLF4on the basis ofcircRNA＿PLEKHM3overexpression, and silencing ofKLF4on the basis ofmiR-320overexpression. qRT-PCR was performed to detect the expression levels of miR-320 in CaSki. Western blot experiments were conducted to determine the expression of KLF4 and EMT markers including E-cadherin, N-cadherin, Vimentin, MMP-2, and MMP-9 in CaSki cells. Transwell assays were performed to measure cell migration and invasion. Results The expression of circRNA＿PLEKHM3 decreased in Hela and CaSki cells(P<0.05), mainly localized in the cytoplasm. The dual luciferase reporter gene experiment demonstrated a targeting relationship betweenmiR-320andcircRNA＿PLEKHM3, as well as betweenKLF4andmiR-320. Overexpression ofcircRNA＿PLEKHM3inhibited the protein expression of miR-320, N-cadherin, Vimentin, MMP-2, and MMP-9, up-regulated the protein expression of E-cadherin, and reduced cell migration and invasion(P<0.05). Overexpression ofmiR-320or silencing ofKLF4on the basis ofcircRNA＿PLEKHM3overexpression both promoted the protein expression of miR-320, N-cadherin, Vimentin, MMP-2, and MMP-9, down-regulated the protein expression of E-cadherin, and increased cell migration and invasion(P<0.05). However, silencing ofKLF4on the basis ofmiR-320overexpression inhibited the protein expression of KLF4, N-cadherin, Vimentin, MMP-2, and MMP-9, up-regulated the protein expression of E-cadherin, and reduced cell migration and invasion(P<0.05). Conclusion Overexpression ofcircRNA＿PLEKHM3regulates EMT in cervical cancer cells through the miR-320/KLF4 axis.