GX1-mediated Liposomes Carrying Adenoviral Vectors For Enhanced Inhibition Of Gastric Cancer Vascular Endothelial Cells

Objective:To seek a gastric cancer targeting drug delivery system of tumor vessels,and improve the specificity and therapeutic effect of the drug,the experiment intended to construct targeting drug delivery system(GX1-AL-Ad5),to study proliferation inhibitory effect of the GX1-AL-Ad5 on the gastric cancer SGC-7901 cells and human umbilical vein endothelial cells(co-HUVEC),to detect the migration inhibiting effect of the GX1-AL-Ad5 by Transwell,and to research the uptake of the gastric cancer SGC-7901 to GX1-AL-Ad5 by confocal laser scanning microscopy.Methods :1.The recombinant adenovirus were amplified by HEK293 cell,purified by CsCl density gradient centrifugation and detected of titer by CPE method.2.The GX1 and PEG2000 were connected by a simple chemical method,and the composite was confirmed by SDS-PAGE electropHoresis and fluorescence intensity.3.Anionic liposomes were prepared by using self-made cholesterol succinate.4.Adenovirus-anionic liposomes(Ad5-AL)were prepared by the method of calcium ion fusion;the functional polypeptide GX1 was connected to theadenovirus anionic liposome(Ad5-AL)by after inserting method and the particle size and Zeta potential of GX1-AL-Ad5 were determined.5.The proliferation inhibition effection of GX1-AL-Ad5 to human gastric cancer cell SGC-7901 was detected by MTT assay;the migration inhibition effection of GX1-AL-Ad5 was investigated by transwell assay;the uptake of the gastric cancer SGC-7901 to GX1-AL-Ad5 was investigated by confocal laser scanning microscope.Results:1.The recombinant adenovirus titer was 2.21×109 pfu/mL.2.The position of the GX1-PEG2000 band was located at 10 kd by SDS-PAGE electropHoresis,which slightly higher than PEG2000.The average connection rate was 75% by detection of fluorescence intensity.3.Anionic liposomes were prepared successfully by using the self-made cholesterol succinate.4.The targeting drug delivery system(GX1-AL-Ad5)was were prepared successfully,its diameter and Zeta potential were all in the optimum range.5.The average proliferation inhibition rate of GX1-AL-Ad5 to SGC-7901 and co-HUVEC cells were 68.36% and 64.13%,which higher than the separate use of GX1-AL or Ad5-AL;the migration inhibition effection of GX1-AL-Ad5 to SGC-7901 cells was significantly higher than that of GX1-AL and Ad5-AL;the uptake of the gastric cancer SGC-7901 to connected GX1 drug was much better than no GX1 drug from results of laser confocal scanning microscope.Conclusion:GX1-mediated liposomes carrying adenoviral vectors could be targeted to gastric cancer vascular endothelial cells and play a role in tumor suppression of proliferation and migration.