Objective:To investigate the feasibility of cyclodextrin-modified calcium phosphate nanoparticles delivering siRNA for the treatment of non-small cell lung cancer cell model A549.Methods:The alendronate-β-cyclodextrin modifier was synthesized chemically.The calcium phosphate nanoparticles (CACaP/siRNA NPs) was prepared and optimized by mixing methods,and the particle size analyzer and gel blocking method were used to determine the stability.The cytotoxicity of CACaP/siRNA NPs and the cell uptake of siRNA in A549 cells were detected by MTT assay and flow cytometer,respectively.Moreover,the expression of EGFR mRNA and protein in A549 cells was detected by RT-PCR and Western-blot,respectively.Results:CACaP/siRNA nanoparticles modified by the alendronate-β3-cyclodextrin were stable and uniform (200 nm and-14 mV),which had good stability and serum protection and could maintain more than 90％ of the cell viability in A549 cells.Meaningfully,it could be transfected into the cells efficiently and down-regulated the expression of EGFR mRNA by 70％,moreover,significantly inhibited the expression of EGFR protein in A549 cells.Conclusion:Cyclodextrin-modified calcium phosphate nanoparticles could effectively deliver siRNA to human lung adenocarcinoma cell line A549,which possesses a better prospect in vivo application.%目的:探讨环糊精修饰的磷酸钙纳米粒递送siRNA用于人非小细胞肺癌细胞A549的可行性.方法:化学合成阿仑膦酸钠-β环糊精修饰物,混合制备并筛选优化包载siRNA的磷酸钙纳米粒CACaP/siR-NA,粒度仪和凝胶阻止法检测其稳定性,MTT法检测其对人肺腺癌细胞系A549生长活性的影响,流式细胞术检测其被肺腺癌细胞系A549细胞摄取的能力,RT-PCR和Western-blot检测其对A549细胞中EGFR表达的影响.结果:成功合成阿仑膦酸钠-β环糊精修饰物,并进一步修饰得到稳定均一的CACaP/siRNA纳米粒,粒径约200 nm,电位约-14 mV,具有良好的放置稳定性和血清保护效果.在A549细胞中可保持90％以上细胞活力,具有良好的安全性,且可以高效地转染入胞,下调70％的EGFR mRNA表达量,显著沉默A549细胞中EGFR蛋白的表达.结论:环糊精修饰的磷酸钙纳米粒可以有效递送siRNA治疗人肺腺癌细胞系A549,具有良好的体内应用前景.
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