〔Abstract〕 Objective To explore the killing effect of chimeric antigen receptor NK-92(CAR-NK-92) cells targeting mesothelin(MSLN) on ovarian cancer cells. Methods The expression of MSLN in primary ovarian cancer tissues and ovarian cancer cell lines was detected by immunohistochemical analysis and immunofluorescence, respectively. The CAR-NK-92 cells targeting MSLN were constructed by lentiviral transfection, the transfection efficiency was detected by flow cytometry, and the killing effect of CAR-NK-92 on ovarian cancer was verifiedin vivoandin vitro. Results MSLN was highly expressed in primary ovarian cancer tissues and ovarian cancer cell lines. Flow cytometry showed that the purity of CAR-NK-92 targeting MSLN was about 70%. The experiments found that MSLN-CAR-NK-92 cells had a strong anti-ovarian cancer effectin vivoandin vitro, and could release more cytokines interferon-γ and tumor necrosis factor-α. Conclusion MSLN-CAR-NK-92 has strong anti-ovarian cancer effectsin vivoandin vitro, providing a new potential treatment option for ovarian cancer patients.