【摘要】：目的:制備裝載靶向Bmi-1基因的siRNA脂質(zhì)復(fù)合物(PILP)并考察其體外對乳腺癌MCF-7細胞增殖的抑制作用、轉(zhuǎn)染效率及對Bmi-1 mRNA表達的影響。方法:以磷脂、膽固醇、DSPE-PEG2000和DSPEPEG2000-maleimide為類脂成分,逆向蒸發(fā)法制備載siRNA的腫瘤靶向脂質(zhì)復(fù)合物。激光納米粒度儀測定脂質(zhì)復(fù)合物的粒徑和電位,凝膠電泳測定其對siRNA的包封率,MTT法檢測脂質(zhì)復(fù)合物對MCF-7細胞體外增殖的抑制,熒光顯微鏡觀察脂質(zhì)復(fù)合物中siRNA被MCF-7細胞攝取的情況,流式細胞儀檢測轉(zhuǎn)染效率,RTPCR法檢測其對MCF-7細胞中Bmi-1 mRNA表達的影響。結(jié)果:制備的載siRNA的腫瘤靶向脂質(zhì)復(fù)合物粒徑為(122.7±1.7)nm、電位為(-22.74±0.96)m V,對siRNA的包封率高;能顯著抑制MCF-7細胞的體外增殖(P0.05),作用72 h抑制率為(55.36±3.5)%;其裝載的siRNA能被MCF-7細胞有效攝取,轉(zhuǎn)染效率為97.07%;能有效沉默MCF-7細胞中Bmi-1 mRNA,沉默效率為69.5%。結(jié)論:采用該方法可制備包封率較高的腫瘤靶向脂質(zhì)復(fù)合物,其介導(dǎo)的siRNA能有效抑制乳腺癌MCF-7細胞的體外增殖、轉(zhuǎn)染效率高、能有效沉默MCF-7細胞中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 鹵1.7) nm, potential (- 22.74 鹵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 鹵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.
【作者單位】： 鄭州大學(xué)附屬鄭州中心醫(yī)院藥學(xué)部;鄭州大學(xué)藥學(xué)院;
【分類號】：R943;R96

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