PLGA纳米颗粒作为基因治疗载体的实验研究

PLGA-Nanoparticles as A Valid Vector for Carrying Antisense Oligonucleotide of Epidermal Growth Factor Receptor in SCCⅦ Cell Lines

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 evaluate the safety and efficacy of Poly（lactide-co-glycolide）（PLGA） nanoparticles（NPs） encapsulating antisense oligonucleotide（Olig） of epidermal growth factor receptor（EGFR） in SCCⅦ cell lines.Methods：PLGA was used to prepare NPs to carry antisense and sense oligonucleotide of EGFR by double-emulsion evaporation technique.The cytotoxic activity of control NPs,antisense-EGFR-NPs,and sense-EGFR-NPs were determined by the MTT colorimetric assay.Expression levels of EGFR mRNA in the SCCⅦ cells which were treated by NPs,was assessed by real-time PCR.Results：The size of designed nanospheres was（116±7.57）nm.Antisense EGFR NPs decreased mRNA levels of EGFR in SCCⅦ cell lines.PLGA-NPs did not show cytotoxicity in all groups.Conclusion：The results show antisense EGFR NPs can efficiently decrease the expression of EGFR,but SCCⅦ cells line has a poor response to antisense-EGFR therapy.These findings suggest that NPs carry antisense oligonucleotide,and could be a valid and non-toxic gene therapy vector to block the target gene.