Abstract: Objective To construct the new targeting delivery system TAT-OSBP-EGFP of human ovarian cancer cell line HO8910，and perform targeted delivery of its properties and in vitro activity identification. Methods The expression vectors of TAT-OSBP-EGFP，TAT-OSBP and OSBP-EGFP via pGEX-6P-3 plasmid were constructed and transfected into E. coli BL21（DE3）. The above vectors were expressed with IPTG induction，purified with GST SefinoseTM Resin colum and subsequently identified by SDS-PAGE and Western blotting analysis. Flow cytometry was used to analyze the cell penetrating efficiency of HO8910 cells which was processed by TAT-OSBP， OSBP-EGFP and TAT-OSBP-EGFP in different concentrations（0，1，5，10μmol/L）. The targeted delivery characteristic of the three fusion proteins for HO8910 cells was observed by fluorescence microscopy in comparison with human colon cancer LoVo cells. The CCK-8 kit was employed to detect the activity of HO8910 cells treated with 0， 1， 5， 10， 15， 40， 60， 80，100μmol/L TAT-OSBP-EGFP for 2h. Results The TAT-OSBP-EGEP expression vector was successfully constructed and a soluble fusion protein was smoothly obtained. Compared with TAT-OSBP，significantly increase was observed in the cell-penetrating rate under 1 and 5μmol/L but not 10μmol/L TAT-OSBP-EGFP to HO8910 cells. The fluorescence intensity（FI） treated by TAT-OSBP-EGFP was higher in HO8910 cells versus LoVo cells. No significant difference was observed on FI between HO8910 cells and LoVo cells treated with TAT-OSBP， while no FI was found in cells treated with OSBP-EGFP. TAT-OSBP-EGEP of different concentrations showed no effect on cell activity（P＞0.05）. Conclusion The carrier system with good human ovarian cell line HO8910 targeting delivery performance is successfully constructed， providing a good foundation of the next tumor-targeting drug delivery vector and tumor targeting killing system.