键合紫杉醇纳米胶束增强替莫唑胺抑制胶质瘤细胞的实验研究

Emperical study of paclitaxel block copolymer nanometer micelle enhances the inhibition effect of temozolomide against malignantglioma

目的在体外研究键合紫杉醇聚合纳米胶束联合替莫唑胺抑制C6胶质瘤的效果和作用机制。方法应用不同浓度的键合紫杉醇聚合胶束及替莫唑胺组合处理C6细胞,应用MTT法观察药物对细胞的增殖抑制作用,流式细胞仪检测细胞凋亡情况,激光共聚焦显微镜检测caspase-3蛋白的表达。结果相对于键合紫杉醇纳米胶束和替莫唑胺药物分单独别作用,1.25～20μg/ml的键合紫杉醇聚合胶束+替莫唑胺组可明显抑制C6胶质瘤细胞的增殖(P<0.01)。且呈明显剂量依赖关系;流式细胞仪检测,键合紫杉醇聚合胶束+替莫唑胺细胞凋亡率增高(P<0.01)。激光共聚焦显微镜检测caspase-3蛋白表达。结果联合药物组与键合紫杉醇纳米胶束、替莫唑胺药物组比较,caspase-3表达明显增加(P<0.01),结论键合紫杉醇聚合胶束可提高替莫唑胺对C6胶质瘤细胞增殖抑制和诱导凋亡作用,且这种作用是通过上调caspase-3蛋白的表达,诱导凋亡实现的。

Objective To investigate the inhibation of C6 glioma cells by paclitaxel block copolymer micelle（PEG- PLA-PTX） and TMZ to find the mechanism of this effect. Methods C6 glioma cell line cultured in vitro were treated by different concentration of PEG-PLA-PTX ,TMZ, and PEC-PLA-PTX ＋ TMZ for 24h-72h. Methyl tiazoy tetrazo li- um （MTT） assay was used to measure the effect of the growth inhibition. Apoptosis of the cells were observed by elec tron microscopy. The expression of caspase-3 in C6 cells were detected by CLSM. Results At does ranges of 1.25,2.5, 5.0,10.0,20.0ug/ml of drugs（PEG-PLA-PTX , TMZ, and PEG-PLA-PTX ＋ TMZ） could inhibit the growth of the cells and present the dependet relationship of concentration in the range of dose. paclitaxel block copolymer micelle can induced apoptosis in c6 cell. Apoptosis is correlated nearly to the block in G2-M phase. The cells apoptosis rate in c6 cells treated by both the PEG-PLA-PTX ＋TMZ was significantly higher than those in C6 cells treated by PEG-PLA- PTX or temozolomide alone （P〈0.01）. The level of caspase-3 expression was significantly higher in C6 cells treated by both the drugs than those in C6 cells treated by PEG-PLA-PTX or temozolomide alone. Conclusion The combinative application of PEG-PLA-PTX and temozolomide can promote the apoptosis and inhibit the proliferation of C6 cells in a synergistic manner. These effects of PEG-PLA-PTX combined with temozolomide may be related with the upregnlation of caspase-3 expression.