载吉西他滨磁性含铁纳米微球治疗胰腺癌细胞的体外研究

In vitro study on gemcitabine carryingg iron-containing magnetic nanoparticles in treating pancreatic carcinoma cells

目的制备载吉西他滨磁纳米微球(Gem-CCMN),考察其协同恒定外磁场下对人胰腺癌细胞CAPAN-2的体外杀伤效应。方法采用化学共价耦联法制备Gem-CCMN,扫描电子纤维镜(SEM)观察纳米粒形态,紫外分光光度计法测定载药量及包封率,体外考察Gem-CCMN在拟溶菌酶存在的不同pH值下药物累积释放率,MTT法检测协同恒定外磁场下Gem-CCMN对CAPAN-2体外增殖的抑制效应,流式细胞术(FCS)检测细胞凋亡。结果 Gem-CCMN呈球形,平均粒径约为50nm,载药量为15%,包封率为45%,pH=7时48h累积释放量仅25%,pH=2时药物累计释放率可达84%;Gem-CCMN协同外磁场时可增强对肿瘤细胞增殖的抑制作用,诱导细胞凋亡和坏死。结论 Gem-CCMN联合外磁场可有效抑制CAPAN-2细胞的生长。

Objective To prepare the gemcitabine carrying magnetic nanoparticles（Gem-CCMN）and to investigate their in vitro killing effects on pancreatic carcinoma cell line CAPAN-2in collaboration of the constant external magnetic fields.Methods Gem-CCMN was prepared by the chemical covalent linkage coupling procedure.Its morphology was observed by scanning electron microscope（SEM）.The drug loading amount and encapsulating ratio were determined by the ultraviolet spectrophotometry.The accumulating release rate of Gem-CCMN under different pH values in the existence of lysozyme was investigated in vitro.The in vitro growth inhibiting effect of Gem-CCMN on CAPAN-2under the collaborative constant external magnetic field was detected by the MTT assay.The apoptosis of CAPAN-2was detected by the flow cytometry.Results Gem-CCMN were spherical with average diameter of 50 nm,the drug loading amount was 15% and the encapsulating ratio was 45%.The 48 hcumulative drug release amount at pH=7was only 25%,but which at pH=7could reach 84%;Gem-CCMN collaborated with external magnetic fields could enhance the killing effect on CAPAN-2,also induced the cellular apoptosis and necrosis.Conclusion Gem-CCMN combined with external magnetic fields could effectively inhibit the growth of CAPAN-2in vitro.