靶向Bmi-1基因的siRNA脂质复合物的制备及体外对乳腺癌MCF-7的抑制作用
发布时间：2019-04-02 19:55
【摘要】：目的:制备装载靶向Bmi-1基因的siRNA脂质复合物(PILP)并考察其体外对乳腺癌MCF-7细胞增殖的抑制作用、转染效率及对Bmi-1 mRNA表达的影响。方法:以磷脂、胆固醇、DSPE-PEG2000和DSPEPEG2000-maleimide为类脂成分,逆向蒸发法制备载siRNA的肿瘤靶向脂质复合物。激光纳米粒度仪测定脂质复合物的粒径和电位,凝胶电泳测定其对siRNA的包封率,MTT法检测脂质复合物对MCF-7细胞体外增殖的抑制,荧光显微镜观察脂质复合物中siRNA被MCF-7细胞摄取的情况,流式细胞仪检测转染效率,RTPCR法检测其对MCF-7细胞中Bmi-1 mRNA表达的影响。结果:制备的载siRNA的肿瘤靶向脂质复合物粒径为(122.7±1.7)nm、电位为(-22.74±0.96)m V,对siRNA的包封率高;能显著抑制MCF-7细胞的体外增殖(P0.05),作用72 h抑制率为(55.36±3.5)%;其装载的siRNA能被MCF-7细胞有效摄取,转染效率为97.07%;能有效沉默MCF-7细胞中Bmi-1 mRNA,沉默效率为69.5%。结论:采用该方法可制备包封率较高的肿瘤靶向脂质复合物,其介导的siRNA能有效抑制乳腺癌MCF-7细胞的体外增殖、转染效率高、能有效沉默MCF-7细胞中Bmi-1 mRNA。
[Abstract]:Aim: to prepare the siRNA lipid complex (PILP) loaded with Bmi-1 gene and investigate its inhibitory effect on proliferation, transfection efficiency and Bmi-1 mRNA expression of breast cancer MCF-7 cells in vitro. Methods: using phospholipid cholesterol DSPE-PEG2000 and DSPEPEG2000-maleimide as lipid compounds siRNA-loaded tumor-targeted lipid complexes were prepared by reverse evaporation. The particle size and potential of lipid complexes were measured by laser nanoparticle analyzer, the encapsulation efficiency of lipid complexes to siRNA was measured by gel electrophoresis, and the inhibition of lipid complexes on the proliferation of MCF-7 cells in vitro was detected by MTT assay. The uptake of siRNA by MCF-7 cells was observed by fluorescence microscope, the transfection efficiency was detected by flow cytometry, and the expression of Bmi-1 mRNA in MCF-7 cells was detected by RTPCR assay. Results: the diameter of siRNA-loaded tumor-targeted lipid complexes was (122.7 卤1.7) nm, potential (- 22.74 卤0.96) m V,), and the encapsulation efficiency of siRNA was high. It could significantly inhibit the proliferation of MCF-7 cells in vitro (P0.05), the inhibition rate was (55.36 卤3.5)% for 72 h, and the siRNA loaded with siRNA could be ingested effectively by MCF-7 cells with a transfection efficiency of 97.07%. The silencing efficiency of Bmi-1 mRNA, in MCF-7 cells was 69.5%. Conclusion: this method can be used to prepare tumor-targeted lipid complexes with high encapsulation efficiency. SiRNA mediated by this method can effectively inhibit the proliferation of breast cancer MCF-7 cells in vitro, and the transfection efficiency is high. It can effectively silence Bmi-1 mRNA. in MCF-7 cells.
【作者单位】： 郑州大学附属郑州中心医院药学部;郑州大学药学院;
【分类号】：R943;R96
[Abstract]:Aim: to prepare the siRNA lipid complex (PILP) loaded with Bmi-1 gene and investigate its inhibitory effect on proliferation, transfection efficiency and Bmi-1 mRNA expression of breast cancer MCF-7 cells in vitro. Methods: using phospholipid cholesterol DSPE-PEG2000 and DSPEPEG2000-maleimide as lipid compounds siRNA-loaded tumor-targeted lipid complexes were prepared by reverse evaporation. The particle size and potential of lipid complexes were measured by laser nanoparticle analyzer, the encapsulation efficiency of lipid complexes to siRNA was measured by gel electrophoresis, and the inhibition of lipid complexes on the proliferation of MCF-7 cells in vitro was detected by MTT assay. The uptake of siRNA by MCF-7 cells was observed by fluorescence microscope, the transfection efficiency was detected by flow cytometry, and the expression of Bmi-1 mRNA in MCF-7 cells was detected by RTPCR assay. Results: the diameter of siRNA-loaded tumor-targeted lipid complexes was (122.7 卤1.7) nm, potential (- 22.74 卤0.96) m V,), and the encapsulation efficiency of siRNA was high. It could significantly inhibit the proliferation of MCF-7 cells in vitro (P0.05), the inhibition rate was (55.36 卤3.5)% for 72 h, and the siRNA loaded with siRNA could be ingested effectively by MCF-7 cells with a transfection efficiency of 97.07%. The silencing efficiency of Bmi-1 mRNA, in MCF-7 cells was 69.5%. Conclusion: this method can be used to prepare tumor-targeted lipid complexes with high encapsulation efficiency. SiRNA mediated by this method can effectively inhibit the proliferation of breast cancer MCF-7 cells in vitro, and the transfection efficiency is high. It can effectively silence Bmi-1 mRNA. in MCF-7 cells.
【作者单位】： 郑州大学附属郑州中心医院药学部;郑州大学药学院;
【分类号】：R943;R96
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1 陈晓梅;王s，
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