Study On The Proliferation, Migration And Drug Resis-tance Of ADM Carried By Single-walled Carbon Nanotubes To Human Ovarian Carcinoma SKOV3/ Admresistant Cells

Objective:To improve the antitumor effect of adriamycin(ADM),preparation of single walled carbon nanotubes with adriamycin(ADM/PEG/SWCNTs) and to investigate the toxicity, drug resistance and migration in human ovarian cancer cell line(SKOV3/ADM). Method:1.The carboxyl group of super high purity single wall carbon nanotube(SWCNTs-COOH) was used as raw material, chemical covalent binding of the double end amino group of polyethylene glycol NH2-PEG-NH22000, and then the physical adsorption of ADM, the preparation of ADM/PEG/SWCNTs complex.2.FTIR was used to characterize the change of surface functional groups of PEG/SWCNTs, and the drug loading was measured by infrared spectroscopy.3.WST-1 was used to detect the inhibitory effect of ADM/PEG/SWCNTs and ADM on SKOV3/ADM at different concentrations;The cell inhibition rate of 24 h,48h,72 h was compared with that of PEG/SWCNTs, ADM, ADM/PEG/SWCNTs at IC50.4.Flow cytometry to detect the change of cell apoptosis after drug induced.5.Hoechst33258 fluorescence staining was observed in the morphological changes of apoptosis.6.The expression of MDR-1m RNA in the cells of each group was detected by the RT-PCR method.7.To extracted the total protein, through the Western blotting method to detect the expression of P-gp protein in each cell.8.Using Transwell assay to detect the migration of SKOV3/ADM under ADM/PEG/SWCNTs drugs in vitro. Results:1.PEG/SWCNTs and ADM/PEG/SWCNTs composite were prepared successfully, 2.the drug loading rate of PEG/SWCNTs was(79.5±10.6)%, and the encapsulation efficiency was(32.0±4.2)%.3.WST-1 experiments showed that IC50 of ADM/PEG/SWCNTs and ADM were 18μg/ml(the concentration of load ADM) and 37.5μg/ml. The same concentration of ADM/PEG/SWCNTs on SKOV3/ADM cells with a significant inhibition, with the prolongation of the time(P < 0.01), PEG/SWCNTs against cancer cells was not obvious(P>0.05).4.Flow cytometry detected ADM/PEG/SWCNTs group and ADM drug group SKOV3/ADM cells apoptosis rate were(14.83±1.86)%VS(5.28±0.45)%,the comparison between the groups(P<0.01), and the difference is statistically significant. 5.The results of hoechst33258 fluorescence staining and flow cytometry were identical, meanwhile the experimental results of WST-1 were verified, which showed that the apoptotic cells were the most in the cellmorphology(F=12.930,P=0.007). 6.The expression of MDR-1m RNA and P-gp protein was down regulated by carbon nanotubes,and analysed by gray scanning,the comparison between the groups,(F=18.634,P=0.003),the difference has statistical significance. 7.ADM/PEG/SWCNTs can inhibit the migration of SKOV3/ADM cells. Conclusion: Preparation of ADM /PEG /SWCNTs nano composite has a high drug loading rate in vitro,which can inhibit the proliferation of human ovarian cancer cell adriamycin resistant strains, and promote the apoptosis of human ovarian cancer adriamycin resistant cells.meanwhile,it can down regulate the expression of MDR-1m RNA and P-gp, and inhibit the migration of ovarian cancer cells.