The invention belongs to the fields of biotechnology and microorganism animal cell lines, and in particular relates to a spontaneous lymphocyte highly metastatic human ovarian sarcomatoid carcinoma cell line, and an establishment method and application thereof. In the invention, the human ovarian sarcomatoid carcinoma cell line SKOV-3 (ATCC code: HTB-77) is taken as a parent cell; and the spontaneous lymphocyte highly metastatic human ovarian sarcomatoid carcinoma cell line SKOV-3/LN403 is obtained by separating and through strategies, such as BALB/c nude mouse intraperitoneal injection, retroperitoneal lymph node continuous screening, amplification in vitro DNA, passage and the like, and the preservation number is CGMCC No.2940. The cell line and an animal model established thereby can simulate the actual clinical situation of a patient with ovarian cancer so as to provide an ideal experiment tool for researching a biological behavior and a regulation and control mechanism for ovarian cancer lymph node metastasis, and screening clinical anti-metastasis medicaments.