Abstract: Objective To investigate the expression of Krüppel-like transcription factor 16 (KLF16) in ovarian cancer tissues and its impact on the proliferation, invasion, and migration abilities of ovarian cancer cells. Methods The expression of KLF16 in ovarian cancer tissues and its correlation with patient prognosis were analyzed by the TCGA database. Immunohistochemistry and real-time quantitative reverse transcription polymerase chain reaction (RT-qPCR) were used to detect the expression of KLF16 in ovarian cancer tissues and normal tissues. Western blot was employed to detect the expression of KLF16 in ovarian cancer cell lines and the effect of KLF16 knockdown in SKOV3 cells. The impact of KLF16 on the proliferation, migration, and invasion of ovarian cancer cells was detected by CCK-8, EdU, colony formation, and Transwell assays. Western blot was used to examine the effect of KLF16 on the expression of epithelial cadherin (E-cadherin), neural cadherin (N-cadherin), and zinc finger protein Snail. Results TCGA database analysis showed that KLF16 was highly expressed in ovarian cancer tissues, and patients with high KLF16 expression showed poorer prognosis (P<0.05). Immunohistochemistry and RT-qPCR results revealed that KLF16 expression in ovarian cancer tissues was significantly higher than in normal tissues (P<0.05). The positive expression of KLF16 was associated with the FIGO stage and histological differentiation of ovarian cancer patients (P<0.05). After KLF16 knockdown in SKOV3 cells, the cells showed significant decreases in proliferation, invasion, and migration abilities, with decreased expression of epithelial-mesenchymal transition (EMT)-related proteins N-cadherin and Snail and increased expression of E-cadherin. Conclusions KLF16 is highly expressed in ovarian cancer tissues and can enhance the proliferation, migration, and invasion abilities of ovarian cancer SKOV3 cells.