BTZ的载双药纳米粒子对PC12细胞增殖抑制作用的研究

Inhibitory effect of PTX and BTZ-loaded nanoparticles on proliferation of PC12 cells

目的研究携载紫杉醇(PTX)和硼替佐米(BTZ)的纳米粒子(NPs)对PC12细胞的增殖抑制作用。方法以PC12细胞为实验对象,采用MTS法检测BTZ、PTX、BTZ-NPs、PTX-NPs以及BTZ-PTX-NPs对PC12细胞的增殖抑制率。用激光共聚焦显微镜观测NPs、PTX-NPs、BTZ-NPs以及BTZ-PTX-NPs的细胞内摄取情况,观测载药纳米粒子的亚细胞定位情况。结果对比给药浓度≤1μg/mL时PC12细胞的存活率(24 h和48 h),MTS实验显示载双药纳米粒子BTZ-PTX-NPs(BTZ∶PTX分别为1∶9、2∶8、3∶7)给药组的存活率(24 h:86.6%、83.8%、83.5%;48 h:74.9%、72.7%、76.2%)均高于BTZ-NPs和PTX-NPs单独给药时的细胞存活率(24 h:47.0%、70.8%;48 h:20.5%、56.9%)(P<0.05)。细胞内摄取实验显示BTZ-NPs和PTX-NPs在PC12内的荧光强度略强于BTZ-PTX-NPs,提示细胞对BTZ-NPs和PTX-NPs的摄取略强于PTX-BTZ-NPs。纳米粒子(绿色荧光)与Lyso Tracker Red DND-99(红色荧光)几乎全部共定位。结论载双药纳米粒子BTZ-PTX-NPs对PC12细胞毒性作用低于载单药纳米粒子PTX-NPs、BTZ-NPs。

Objective Study on the inhibitory effect of paclitaxel and bortezomib loaded nanopaticles on proliferation of PC12 cells.Methods PC12 cells were used as experimental subjects.MTS assay was used to detect the proliferation inhibition rate of PC12 cells by BTZ,PTX,BTZ-NPs(bortezomib nano paticles),PTX-NPs(paclitaxel nano paticles)and BTZ-PTX-NPs.The intracellular uptake of NPs,PTX-NPs,BTZ-NPs and BTZ-PTX-NPs was observed by laser scanning confocal microscopy.The subcellular localization of drug-loaded nanoparticles was observed.Results The survival rate of PC12 cells(24 h and 48 h)was compared with the concentration of≤1μg/mL.The MTS assay showed that the survival rates of double-drug nanoparticles BTZ-PTX-NPs(BTZ:PTX is 1∶9,2∶8,3∶7 respectively)administered group(24 h:86.6%,83.8%,83.5%;48 h:74.9%,72.7%,76.2%)was higher than those of BTZ-NPs and PTX-NPs when administered alone(24 h:47.0%,70.8%;48 h:20.5%,56.9%)(P<0.05).Intracellular uptake experiments showed that the fluorescence intensity of BTZ-NPs and PTX-NPs in PC12 was slightly stronger than that of BTZ-PTX-NPs,suggesting that the uptake of BTZ-NPs and PTX-NPs by cells was slightly stronger than that of BTZ-PTX-NPs.Nanoparticles(green fluorescence)and Lysotracker Red DND-99(red fluorescence)were almost all co-localized.Conclusion The cytotoxicity of double-drug loaded nanoparticles BTZ-PTX-NPs on PC12 cells was lower than that of single-drug loaded nanoparticles PTX-NPs and BTZ-NPs.