顺铂联合米托蒽醌调节脑胶质瘤U87细胞Sonic Hedgehog信号通路的研究

目的：观察米托蒽醌（Mitoxantrone,MXT）联合顺铂（Cisplatin,CDDP）对脑胶质瘤U87细胞杀伤活性及对Sonic Hedgehog信号通路的影响。方法：应用MTT法检测不同浓度米托蒽醌、顺铂以及两药物联合对U87细胞成活率的影响。显微镜观察细胞的形态变化DiOC6荧光染料对细胞线粒体染色检测其膜电位变化来反映细胞凋亡。RT-PCR法检测顺铂、米托蒽醌及两药联合对U87细胞Gli1和Ptch基因表达的影响。结果：MTT结果显示顺铂、米托蒽醌均可以有效抑制U87细胞的增殖,当米托蒽醌和顺铂浓度≤0.625μg/ml时,两药联合对U87细胞增殖具有协同抑制作用;细胞形态变化及线粒体膜电位结果显示,单药处理可促进U87细胞凋亡,而联合用药可以协同促进U87细胞的凋亡;RT-PCR法检测显示顺铂对Gli1基因的表达有上调作用,而米托蒽醌、米托蒽醌联合顺铂能下调Ptch和Gli1基因的表达。结论：胶质瘤U87细胞在化疗药物米托蒽醌以及两药物联合作用下可影响Sonic Hedgehog信号通路,协同发挥其促凋亡作用,进而增强肿瘤细胞对化疗药物的敏感性。

Effect of mitoxantrone combined with cisplatin on Sonic Hedgehog signal pathway of glioma cell line U87

Objective：To observe the effect of mitoxantrone（MXT） combined with cisplatin（CDDP）on Sonic Hedgehog signal pathway of glioma cell line U87 in vitro.Methods：The survival rate of U87 cells after treatment with different concentrations of MXT,CDDP and the same concentrations of MXT plus CDDP were observed by MTT assay.Using DiOC6 dye staining to detect the mitochondria membrane potential changes in U87 cells and morphological changes were observed the apoptosis.The expression of Gli1 and Ptch gene on U87 cells were detected by RT-PCR.Results：MTT and mitochondrial membrane potential assay indicated that CDDP at low concentrations（≤0.625μg /ml） combined with MXT can significantly enhance the inhibitory effect and apoptosis of U87 cells.RT-PCR assay showed that CDDP increased the expression of Ptch and Gli1 gene,but MXT and MXT＋CDDP can decrease the Ptch and Gli1 gene expression.Conclusion：U87 glioma cells treated with MXT alone or treated with MXT combined with CDDP can affect the Sonic Hedgehog signaling pathway,which played an important role in enhancing the apoptosis,thus enhancing the sensitivity of U87 cells to chemotherapeutic drug.