Experimental Study On Anti-hepatoma Cells Treated With Arsenic Trioxide Loaded With Microspheres

Purpose Preparation of arsenic trioxide(ATO)drug-eluting beads and study their sustained-release characteristics,hepatoma cell lines Hep G2,MHCC97 H as the study object,from the cell proliferation,apoptosis,invasion and VEGF,MMP9 m RNA expression levels and other aspects,to investigate the effect of ATO and ATO-loaded microspheres on hepatoma cells in vitro,it lays a theoretical foundation for animal experimental studies in the future,in order to provide new ideas and theoretical basis for the comprehensive interventional treatment of liver cancer.Materials and Methods1.Preparation of ATO drug-eluting beads in vitro,calculate the loading efficiency(LE%)and entrapment efficiency(EE%)and the drug release experiments were carried out to understand the sustained release of ATO drug-eluting beads;2.The proliferation of Hep G2 and MHCC97 H cells were detected by CCK8 at different times(0,12,24,48,72,96h)Inhibition of proliferation;3.Invasion experiments were performed to observe the effects of different concentrations(0?5?10?15?20ng/ml)of ATO and ATO loaded microspheres on the invasion of Hep G2 and MHCC97 H cells after treatment for 48 h.4.Double staining was used to detect the effects of ATO and ATO drug-eluting beads at different concentrations(0,5,10,15,20 ng/ml)on the apoptosis of Hep G2 and MHCC97 H cells 48h;5.The expression of VEGF and MMP9 m RNA in Hep G2 and MHCC97 H cells treated with different concentrations(0,5,10,15,20 ng/ml)of ATO and ATO drug-eluting beads for 24 h and 48 h were detected by RT-PCR.6.Western Blot was used to detect the expression of VEGF protein in Hep G2 and MHCC97H cells treated with different concentrations(0,5,10,15,20 ng/ml)of ATO and ATO drug-eluting beads for 24 h and 48 h.Results1.Calli Spheres? drug-eluting beads(100-300?m)on ATO the best time for drug loading 40min;1g particle size 300-500?m drug-eluting beads,into 60mg(10mg/ml,6ml)of ATO,after mixing 40 min,Can detect the internal energy load13.8±1.49 mg ATO;entrapment efficiency of 25.0±1.34%;2.Calli Spheres? ATO drug-eluting beads(100-300?m)showed a burst release within 30 minutes,and the burst release amounted to 31.42% of the total drug release,and the release amount reached 47.17% at 48 hours;3.At the same time,ATO group and ATO drug-eluting beads group,the concentration of 0~20ng/ml range,as the concentration of ATO increased,the proliferation inhibition rate of Hep G2 and MHCC97 H cells increased gradually,There was a statistically significant difference within each group(P<0.05).In ATO group and ATO drug-eluting beads group,in the same concentration,the proliferation inhibition rate of Hep G2 and MHCC97 H cells increased gradually with the prolonged action time,but there was no significant difference between the two groups(P>0.05);4.ATO drug-eluting beads,like single-agent ATO,significantly inhibited the invasive ability of Hep G2 and MHCC97 H cells with increasing concentrations.5.The ATO drug-eluting beads with the same concentration could promote the apoptosis of Hep G2 and MHCC97 H cells at the same time compared with the single drug ATO;6.The same concentration of ATO drug-eluting beads can also inhibit the levels of vascular endothelial growth factor(VEGF)and matrix metalloproteinase 9(MMP9)in the same timecompared with the single drug ATO.Conclusion1.Calli Spheres drug-eluting beads(100-300?m)can be loaded with a certain amount of positively charged ATO,having a sustained-release capacitythe;2.ATO drug-eluting beads with the same concentration can inhibit the proliferation of Hep G2 and MHCC97 H cells and promote the apoptosis of Hep G2 and MHCC97H cells at the same time compared with the single drug ATO.3.The same concentration of ATO drug-eluting beads could also inhibit VEGF and MMP9 expression at the same time compared with the single drug ATO.