制备纳米粒联合高强度聚焦超声治疗三阴性乳腺癌:体外实验

Preparation of nanoparticle combining with high intensity focused ultrasound for treating three negative breast cancer:In vitro experiment

目的制备特异性靶向三阴性乳腺癌(TNBC)纳米粒(NP)AS1411/PFH-NP,评估其联合高强度聚焦超声(HIFU)治疗TNBC的价值。方法以双乳化法将十四氟己烷(PFH)装载于PLGA@PEG,以AS1411核酸适配体与PFH-NP连接,制备AS1411/PFH-NP,观察其物理特性、靶向能力及体外超声显像能力,评价HIFU联合AS1411/PFH-NP消融离体牛肝组织及杀伤MB-231 TNBC细胞的价值。结果成功制备AS1411/PFH-NP,其形态较佳,对MB-231 TNBC细胞具有良好靶向能力。AS1411/PFH-NP组CEUS图像灰度值大于磷酸盐缓冲液(PBS)组及NP组(t=17.24、5.60,P均<0.05)。经HIFU+AS1411/PFH-NP消融后,牛肝组织凝固性坏死体积大于经HIFU+PBS(t=7.99,P<0.01)及HIFU+NP消融后(t=4.33,P<0.05);且HIFU+AS1411/PFH-NP消融能效因子显著低于HIFU+PBS(t=5.41,P<0.01)及HIFU+NP消融(t=4.82,P<0.01)。HIFU+AS1411/PFH-NP组MB-231细胞凋亡率高于其他组(P均<0.05)。结论特异性靶向TNBC AS1411/PFH-NP可提高HIFU体外消融效率。

Objective To prepare nanoparticle(NP)AS1411/PFH-NP that specifically targeting triple negative breast cancer(TNBC),and to observe the value of AS1411/PFH-NP combining with high intensity focused ultrasound(HIFU)for treating TNBC.Methods The liquid fluorocarbon(PFH)was loaded into PLGA@PEG using double emulsification method,and the aptamer AS1411 was connected with PFH-NP to prepare AS1411/PFH-NP.The physical properties,targeting ability and ultrasonic imaging ability of AS1411/PFH-NP were observed.The value of HIFU combining with AS1411/PFH-NP for ablation of isolated bovine liver and killing MB-231 TNBC cells were analyzed.Results AS1411/PFH-NP was successfully prepared,which had good morphology and good targeting ability to MB-231 TNBC cells.The gray value of CEUS of AS1411/PFH-NP group was higher than that of phosphate buffer solution(PBS)group and NP group(t=17.24,5.60,both P<0.05).The volume of coagulation necrosis in bovine liver tissue after HIFU+AS1411/PFH-NP ablation was larger than that after HIFU+PBS(t=7.99,P<0.01)and HIFU+NP(t=4.33,P<0.05)ablations,and the energy efficiency factor of HIFU+AS1411/PFH-NP ablation was significantly lower than that of HIFU+PBS(t=5.41,P<0.01)and HIFU+NP(t=4.82,P<0.01)ablations.The apoptosis rate of MB-231 cells of HIFU+AS1411/PFH-NP group was higher than that of the other groups(all P<0.05).Conclusion AS1411/PFH-NP that specifically targeting TNBC could improve the ablation efficiency of HIFU in vitro.