| Objective: To explore a kind oftargeted,little side effects,both with diagnosis and treatment of drug-resistant treatment of head and neck squamous cell carcinoma.Methods: Gefitinib liposomes and chloroquine liposomes were prepared by passive drug loading and active drug loading respectively.The ratio of different lipid components was changed so that the encapsulation efficiency of the two liposomes was as high as possible.Ultrasound microbubbles were prepared,and the prepared liposomes were connected to the microbubbles by means of biotin-avidin.The connection of the two was observed under a fluorescence microscope.The sensitivity of CAL-27,HN-4 and HN-30 to the treatment of gefitinib alone,chloroquine alone,and combination of gefitinib plus chloroquine was assessed in vitro and choose the most sensitive cell line to the combination therapy.The sensitive strain cells were divided into 6 groups: PBS buffer+US,GE-Lipo+ CQ-Lipo+US,MB-GE-Lipo+US,MB-CQ-Lipo+US,MB-CQ-Lipo+US,MB-GE-Lipo + MB-CQ-Lipo,MB-GE-Lipo + MB-CQ-Lipo(with ultrasound).Using the MTS kit to detect the inhibitory effect of proliferation on each group.Results :When EPC:Chol=5:1,the entrapment efficiency of gifitinib-loaded liposomes was(49.7±5.9)%,and the entrapment efficiency was significantly different from the 2:1 group.The particle size was(74.55±0.80).)nm,and the PDI<0.2.There was no significant difference in the entrapment efficiency between the 2:1 and 8:1 groups(P>0.05).Compared with 10:1,20:1,there was significant difference in the entrapment efficiency of 2:1(P﹤0.05).Group 5:1,8:1,10:1,and 20:1 compared with each other,but there was no significant difference between every two groups(P>0.05).The average particle size of the chloroquine loaded liposomes was(68.94±8.18)nm,the zeta potential was(-2.04±0.18)m V,PDI<0.2,and the encapsulation efficiency was(92.33±2.52)%.Under the fluorescence microscope,there was a circle of green fluorescence around the microbubbles.In vitro experiments of three kinds of cells,when the concentration of gefitinib was IC50 and the concentration of chloroquine was 50μg/ml,the inhibition efficiency of CAL-27 cells and HN-30 cells was statistically significant(P <0.05).When HN-30 cells and HN-4 cells was treated with the concentration of IC50 of gefitinib and50μg/ml of chloroquine,there was no significant difference in the inhibition of cell proliferation after combination therapy(P>0.05).When chloroquine is below 25 μg/ml,the proliferation inhibition rate of both cells was statistically significant(P<0.05).For the HN-30 cells,between MB-GE-Lipo + MB-CQ-Lipo(with ultrasound)group and the PBS+US group,there are statistically significant differences(P<0.05),and there was a statistically significant difference between(MB-GE-Lipo+ MB-CQ-Lipo,with US)groupand(MB-GE-Lipo+ MB-CQ-Lipo)group.(P<0.05).There was no significant difference between(MB-GE-Lipo+ MB-CQ-Lipo,with US)group and MB-GE-Lipo+US group(P>0.05).Conclusions: When EPC:Chol=5:1,the EE of GE-Lipo was higher,and the ratio of cholesterol was appropriate,and the liposome was more stable.Liposomes and microbubbles can be linked through biotin-avidin.Compared to Cal-27 cells and HN-4 cells,HN-30 cells were more sensitive to the combination therapy of gefitinib and chloroquine.(MB-GE-Lipo+ MB-CQ-Lipo,with US)group has significant inhibition effect on tumor proliferation. |
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