Studies Of Multilayer Microspheres Loaded With Paclitaxel And 5-fluorouracil On Biological Behaviour In Human Carcinoma Cells Tca8113 And BcaCD885

Objective:The microspheres were prepared in a sterile environment based on the methods from previous studies by using alginate and chitosan as a drug delayed release carriers.The microspheres loaded or unloaded with the paclitaxel and 5-fluorouracil prqpared were co-cultured with oral squamous carcinoma cell Tca8113 and BcaCD885 seperately to observe cell morphology,proliferation,cloning,wound healing,invasion,migration and apoptosis.Explore the anti-tumor effect of multi-drug loaded sustained-release microspheres and lay a foundation for its clinical application.Methods:We prepared multilayer microspheres in a sterile environment to study the effects on biological behavior in human cheek well differentiated squamous carcinoma cell line(BcaCD885)and human tongue poorly differentiation squamous carcinoma cell line(Tca8113).Sterile microspheres and cells were co-cultured to observe changes in cells under a microscope.Cell proliferation was tested by MTT assay.The ability of cell clone formation was detected by cell clone formation assay.The effects of microspheres on cell migration and invasion were examined by wound healing assay and Transwell- assay.Cell apoptosis and necrosis were analysed by Annexin V/PI staining and flow cytometry.Results:1.Multilayer loading paclitaxel and 5-fluorouracil microspheres and multilayer non-loaded microspheres were prepared.They were spheroid or ellipsoid of uniform size and their mean particle size were(815.93±15.47)?m.2.The morphology of oral squamous carcinoma cells co-cultured with drug-loaded microspheres changed significantly.It could be observed that BcaCD885 cells changed from round or oval shape into irregular shape,and finally the cells dispersed.Also,Tca8113 cells changed from polygon into long spindle,later the cells were broken and the morphology almost disappeared.Nevertheless,the cells co-cultured with microspheres unloaded drugs maintained their original morphology.3.After co-cultured with drug-loaded microspheres,the proliferation abilities of BcaCD885 and Tca8113 cells were obviously inhibited(P<0.01).Results of clonal formation experiments showed that drug-loaded microspheres had a significant decrease on cell proliferation and clonal formation(P<0.001).4.It was observed that the ability of migration and invasion for Tca8113 and BcaCD885 cells were significantly decreased after co-culturing with drug-loaded microspheres(P<0.05).5.It was found that drug-loaded microspheres significantly promoted the apoptosis of Tca8113 and BcaCD885 cells(P<0.05),while non-drug-loaded microspheres had no significant effect.Conclusions:1.Multilayer drug-loaded microspheres containing paclitaxel and 5-:fluorouracil and unloaded multilayer microspheres were successfully prepared using drug delayed release carrier produced by alginate and chitosan under sterile conditions.They were spheroid or ellipsoid of uniform size and their mean particle size were(815.93±15.47)?m.2.The prepared multi-drug loaded microspheres can not only make the morphology of oral squamous carcinoma cell lines Tca8l13 and BcaCD885 tend to be irregular or even broken,but also inhibit their abilities of proliferation,cloning,migration and invasion,together with promoting cell apoptosis.