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目的:用针对表皮生长因子受体(epidermal growth factor receptor,EGFR)的小分子多肽配体D4修饰PEG化的阳离子脂质体,观察其提高质粒DNA和siRNA对肿瘤细胞转染效率的作用。方法:将D4连接在DSPE-PEG2000的末端以修饰PEG化的阳离子脂质体,检测该载体系统对高表达EGFR的人非小细胞肺癌细胞株H1299中质粒DNA转染效率的影响,Sirius照度仪检测质粒DNA转染后H1299细胞荧光素酶的表达,荧光显微镜观察转染FAM-siRNA后H1299细胞的荧光强度。结果:制备的脂质体/质粒DNA复合物随着电荷比的提高,复合物粒径逐渐减小,复合物Zeta电位逐渐升高。在质粒DNA的转染中,与无修饰的非靶向脂质体相比,D4修饰的脂质体可以显著提高H1299细胞中荧光素酶的表达(P<0.05或P<0.01);D4修饰的脂质体在各个电荷比处对H1299细胞的转染效率显著高于无修饰的非靶向脂质体(P<0.05或P<0.01)。在FAM-siRNA的转染中,荧光显微镜下可以观察到D4修饰的脂质体组有更高水平的FAM荧光强度。结论:D4修饰的阳离子脂质体提高高表达EGFR肿瘤细胞中质粒DNA和siRNA的转染效率。
OBJECTIVE: To modify PEGylated cationic liposomes with small molecule peptide ligand D4 targeting epidermal growth factor receptor (EGFR), and to observe its effect on enhancing transfection efficiency of tumor cells by plasmid DNA and siRNA. METHODS: D4 was attached to the end of DSPE-PEG2000 to modify PEGylated cationic liposomes. The effect of this vector system on the transfection efficiency of plasmid DNA in human non-small cell lung cancer cell line H1299 with high expression of EGFR was examined. The Sirius illuminance The expression of luciferase in H1299 cells transfected with plasmid DNA was detected, and the fluorescence intensity of H1299 cells transfected with FAM-siRNA was observed by fluorescence microscopy. Results: With the increase of the charge ratio, the particle size of the prepared liposome / plasmid DNA composite decreased and the Zeta potential of the complex increased gradually. D4-modified liposomes significantly increased luciferase expression in H1299 cells (P <0.05 or P <0.01) compared with unmodified non-targeted liposomes in transfection of plasmid DNA; D4 modification The transfection efficiency of H1299 cells at various charge ratios was significantly higher than that of unmodified non-targeted liposomes (P <0.05 or P <0.01). In FAM-siRNA transfection, a higher level of FAM fluorescence intensity was observed in the D4 modified liposomes under a fluorescence microscope. Conclusion: D4-modified cationic liposomes can improve the transfection efficiency of plasmid DNA and siRNA in EGFR tumor cells.