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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。
Objective To investigate the chemosensitivity of stathmin gene silencing in combination with docetaxel (Taxotere) to osteosarcoma cells. Methods siRNA eukaryotic plasmids targeting stathmin mRNA were successfully transfected into LM8 osteosarcoma cells by lipofectamine and stable cell lines were obtained. Taxus and cisplatin induced the control LM8 cells and transfected LM8 cells respectively. The growth of the above two groups of cells was determined by trypan blue staining. The proliferation of single cells in two groups was observed by soft agar colony formation assay. MTT Method was used to test the inhibitory concentration of Taxus and cisplatin on the two groups of cells. Flow cytometry was used to quantitatively analyze the effect of Taxox on the cell cycle and the apoptosis rate of the two groups. Results Compared with the control group, Taxus and cisplatin all inhibited the growth of LM8 cells, but the inhibitory effect of Taxotere on the transfected LM8 cells was more obvious. Taxus was more significantly reduced than cisplatin In vitro proliferation activity of single cells, especially the transfection of LM8 osteosarcoma cells; Taxus and cisplatin decreased the half inhibitory concentration of transfection of LM8 cells compared with the control group, but decreased the concentration of Taxotere half-value significantly; Cytometry showed that Taxotere could more easily block the transfection of LM8 cells in the G2 / M phase and more easily induce the apoptosis of the transfected LM8 cells. Conclusion The stathmin gene silencing can significantly increase the chemosensitivity of Taxotere to osteosarcoma LM8 cells and induce apoptosis in combination with it, providing a new idea for the treatment of tumors.