Construction and biological activity identification of new human ovarian cancer epithelial cell HO8910 tar-geting delivery system

LIAO Hua; KANG Jiali; JIANG Wenyan; WANG Xiaoxia; ZHONG Jiali; ZHOU Cong

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-EGFP and OSBP-EGFP via pGEX-6P-3 plasmid were constructed and transfected into E. coli BL21(DE3). The a-bove 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-EGFP, OSBP-EGFP and TAT-OSBP-EGFP in different concentrations(0, 1, 5, 10μmol/L). The targeted delivery character-istic of the three fusion proteins for HO8910 cells was observed by fluorescence microscopy in comparison with human colon cancer Lo-Vo 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-EGFP, 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-EGFP , 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 suc-cessfully constructed, providing a good foundation of the next tumor-targeting drug delivery vector and tumor targeting killing system.