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目的:探讨PLGA材料构建的纳米粒载体导入表皮生长因子受体(EGFR)反义寡核苷酸在头颈鳞癌基因治疗中的可行性,为头颈肿瘤基因治疗中载体的选择提供一个新的研究思路。方法:以PLGA为材料,采用油包水双乳化溶剂蒸发法制备载EGFR正义、反义寡核苷酸纳米颗粒;纳米颗粒转染SCCⅦ细胞株;MTT法了解纳米颗粒对细胞的毒性;通过实时荧光定量PCR检测转染后EGFR基因mRNA表达水平。结果:获得了制备载寡核苷酸PLGA纳米颗粒工艺流程,PLGA纳米颗粒平均粒径116 nm±7.57 nm。纳米颗粒体外转染SCCⅦ细胞,MTT结果显示纳米颗粒对细胞生长无明显抑制效应,同时具有明显抑制EGFR基因mRNA表达效应。结论:PLGA纳米颗粒可以有效地载入反义寡核苷酸,达到抑制靶基因的效果,同时无明显的细胞毒性。
OBJECTIVE: To investigate the feasibility of using nanoparticle-loaded PLGA-loaded antisense oligonucleotide into epidermal growth factor receptor (EGFR) antisense oligonucleotide in the gene therapy of head and neck squamous cell carcinoma and to provide a new study for the selection of vector in gene therapy of head and neck cancer Ideas. Methods: PLGA as a material, double-emulsion with water-in-oil solvent evaporation prepared EGFR sense and antisense oligonucleotide nanoparticles; Nanoparticles transfected SCCⅦ cell line; MTT method to understand the nanoparticles on the cell toxicity; through real-time Fluorescent quantitative PCR was used to detect the mRNA expression of EGFR gene after transfection. Results: The process of preparation of oligo-loaded PLGA nanoparticles was obtained. The average diameter of PLGA nanoparticles was 116 nm ± 7.57 nm. Nanoparticles transfected SCCⅦ cells in vitro, MTT results showed that nanoparticles have no significant inhibitory effect on cell growth, but also significantly inhibited EGFR gene mRNA expression. Conclusion: PLGA nanoparticles can effectively load antisense oligonucleotides to inhibit the target gene without obvious cytotoxicity.