Abstract: Objective :To isolate and characterize cancer cell subclones with different metastatic potential from human ovarian cancer cell line. Methods :Three sublines with different metastatic potential from human ovarian cancer cell line一SIOV3 (S1, S6andS14) were established, using cell clone technique and cell electrophoresis. The invasiveness and migratory capacity among the three subgroups were observed by vitro growth, Matrigel invasion assay and in viva tumorigenicity and spontaneous metastasis assay in nude mice. Results :Among the three subclones, S1 expressed as the highly metastatic phenotype, with 70.00% metastasis frequency 6 weeks after subcutaneous inoculation into nude mice, whereas S14 was not metastatic. In vitro, S1 was found to be the most highly invasive cell line in Matrigel invasive assay, while S14 was found to have the least invasive ability. Conclusions :Successful establishment of these subclones with different metastatic potential may be valuable for further study on the molecular mechanisms of cancer metastasis and the cloning of related genes of cancer metastasis.