载紫苏醇线粒体靶向脂质体(POH/DPT-LN)对乳腺癌MDA-MB-231细胞的作用研究

Effect of perillyl alcohol-loaded mitochondria-targeted liposomes(POH/DPT-LN)on breast cancer MDA-MB-231 cells

目的:制备载紫苏醇(POH)二硬脂酰基磷脂酰乙醇胺(DSPE)-聚乙二醇(PEG)-(3-丙羧基)三苯基溴化磷(TPP)脂质体(POH/DPT-LN),考察其理化特征、线粒体靶向性及对乳腺癌MDA-MB-231细胞的抑制作用。方法:采用薄膜分散法制备POH/DPT-LN,采用CCK-8、细胞划痕实验、Transwell、细胞凋亡实验、JC-1染色实验及western blotting实验对该载药系统线粒体靶向性及抑瘤作用进行评价。结果:POH/DPT-LN外观形态呈均一、完整的类球形,平均粒径为(141.0±0.8)nm,zeta电位为(29.5±2.8)m V,PDI为(0.185±0.006),包封率及载药量分别为(88.6±1.8)%和(17.7±0.3)%;POH/DPT-LN的IC50为63.32μg/mL,其对MDA-MB-231细胞生长抑制效果良好;与POH单药组相比较,POH/DPT-LN组的MDA-MB-231细胞迁移能力显著减弱,抑制细胞侵袭效果明显(P<0.01);POH/DPT-LN组使得线粒体内POH蓄积浓度,对细胞杀伤效果增强(P<0.01)。POH/DPT-LN可明显提高促凋亡P53、Bax、Caspase-3蛋白表达及降低抗凋亡Bcl-2蛋白表达。结论:POH/DPT-LN具有较高的包封率及载药量,能够将POH药物靶向传递至线粒体内,更有效的抑制MDA-MB-231乳腺癌细胞的增殖,增强药物抗肿瘤效果。

Objective:To prepare perillyl alcohol(POH)-loaded distearoyl phosphoethanolamine(DSPE)-polyethylene glycol(PEG)-(3-carboxy propyl)triphenyl phosph-onium bromide(TPP)-liposomes(POH/DPT-LN),and to investigate the physicochemical characteristics,mitochondrial targeting and the inhibitory effect on breast cancer MDA-MB-231 cells.Methods:POH/DPT-LN was prepared by thin film dispersion method.Cell counting kit-8(CCK-8)assay,cell scratch experiment,Transwell,apoptosis experiment,JC-1 staining experiment and western blotting were used to evaluate the mitochondrial targeting and tumor-inhibitory effects of drug-loading systems.Results:POH/DPT-LN was uniform and round,with an average particle size of(141.0±0.8)nm,zeta potential of(29.5±2.8)mV,PDI of(0.185±0.006),encapsulation efficiency and drug loading capacity of(88.6±1.8)%and(17.7±0.3)%,respectively.The IC50 of POH/DPT-LN was 63.32μg/mL,and it had a good inhibitory effect on the growth of MDA-MB-231 cells.Compared with the POH monotherapy group,the migration ability of MDA-MB-231 cells in the POH/DPT-LN group was significantly weakened,and the effect on inhibiting cell invasion was obvious(P<0.01).POH/DPT-LN group increased the accumulation concentration of POH in the mitochondria and enhanced the cell killing effect(P<0.01).POH/DPT-LN can significantly increase the protein expression of pro-apoptotic P53,Bax,Caspase-3 and reduce the protein expression of anti-apoptotic Bcl-2.Conclusion:POH/DPT-LN has good encapsulation efficiency and drug loading capacity.It can target POH drugs into mitochondria,more effectively inhibit the proliferation of MDA-MB-231 breast cancer cells,and enhance the anti-tumor effect of drugs.