摘要
目的探讨stathmin基因沉默协同增效多烯紫杉醇(泰索帝)对骨肉瘤细胞的化疗敏感性。方法将已构建成功的针对stathmin mRNA的siRNA真核质粒经脂质体转染LM8骨肉瘤细胞并获得稳定的细胞系。泰索帝和顺铂分别诱导对照LM8细胞、转染的LM8细胞,台盼蓝染色法测定以上2组细胞的生长,软琼脂集落形成实验观察2组细胞的单个细胞体外增殖活力,噻唑蓝MTT法分别检测泰索帝和顺铂对2组细胞半数抑制浓度,流式细胞术定量分析泰索帝对2组细胞周期及细胞凋亡率的影响。结果与对照组相比,不同浓度的泰索帝和顺铂均抑制了LM8细胞的生长,但泰索帝对转染的LM8细胞生长抑制作用更为明显;泰索帝较顺铂更加明显减少了单个细胞,特别是转染LM8骨肉瘤细胞的体外增殖活力;泰索帝和顺铂对转染的LM8细胞半数抑制浓度均较对照组下降,但泰索帝半数抑制浓度下降更显著;流式细胞术检测显示,泰索帝更易使转染LM8细胞阻滞于G2/M期,更易诱导转染LM8细胞的凋亡。结论stathmin基因沉默能明显增加泰索帝对骨肉瘤LM8细胞的化疗敏感性并协同诱导细胞凋亡,为肿瘤的治疗提供了一种新思路。
ObjectiveTo investigate the enhancive effect of small interfering RNA（siRNA）targeting stathmin gene on sensitivity of LM8 cells to docetaxel.MethodsThe siRNA eukaryotic express vector targeting stathmin gene was successfully constructed and transduced into LM8 osteosarcoma cells by liposomal trasfection.The stable transfected LM8 osteosarcoma cell strain was gained.The controlled LM8 cells and transfected LM8 cells were induced by docetaxel and platinol,respectively.The cellular growth rate was assayed by trypan-blue staining.The cellular growth activities were assayed by clonogenic assays in soft agar culture.The 50% inhibiting concentrations（IC50）of docetaxel and platinol were measured by MTT colorimetry.The cell cycle and apotosis rate which were influenced by docetaxel were analyzed by flow cytometry.ResultsDocetaxel and platinol with different concentrations all inhibited cellular growth in two groups,but it was more obvious for docetaxel in inhibiting growth of transfected LM8 cells;clonogenicity of cells with docetaxel and platinol were reduced in two groups,but it was drastical for docetaxel to reduce clonogenicity of transfected LM8 cells in soft agar culture.The 50% inhibiting concentrations（IC50） of docetaxel and platinol on transfected LM8 cells were lower than that on controlled LM8 cells,but reduction of IC50 of docetaxel was more obvious.Comparing with control group,the percentage of G2/M phase and apoptosis rate of transfected cells which were induced by docetaxel were all higher.ConclusionsStathmin gene silencing could improve sensitivity of LM8 cells to docetaxel and synergistically induce cellular apoptosis.Combinating siRNA technique with docetaxel medication might provide a novel and attractive strategy for clinical therapy of osteosarcoma.
出处
《中国临床研究》
CAS
2010年第3期181-183,I0001,I0002,共5页
Chinese Journal of Clinical Research
