Abstract: 【Abstract】 Objective To investigate the role of Fibronectin in the formation of multi-cellular spheroid of ovarian cancer and the integrin receptor involved in the process. Methods In vitro model of multi-cellular spheroid of SKOV3 was constructed by liguid overlay technique. The influence of fibronectin on the formation of the spheroid was observed. The gene expressions of potential integrin receptors were examined from the levels of mRNA and protein using real time reverse transcription PCR and Western-blot. Results Fibronctin stimulated the formation of multi-cellular spheroid of ovarian cancer larger than 250 μm. fibronectin suppressed the expression of subtype of integrin receptor ITGA5. Conclusion Fibronectin can enhance the formation of multi-cellular spheroid of ovarian cancer. The subtype of integrin receptor ITGA5 is probably involved in the process.