论文部分内容阅读
目的:通过转染可抑制Tca-8113细胞tankyrase 1表达的siRNA,达到抑制Tca-8113细胞增殖的目的。方法:采用体外转录合成方法,合成针对tankyrase 1的siRNA,并进行Cy-3荧光标记,用脂质体转染至Tca-8113细胞,荧光显微镜观察判断转染效果。应用噻唑蓝(MTT)法和流式细胞仪检测siRNA的作用效果;应用Western blot方法鉴定siRNA抑制tankyrase 1表达效果。结果:tankyrase 1 siRNA组与空白对照组、脂质体对照组相比,可诱导Tca-8113细胞tankyrase 1表达下调,Western blot结果分析显示,转染siRNA后可下调tankyrase 1表达量的1/2,并抑制Tca-8113细胞的增殖(P<0.05),同时改变了细胞周期各时相的细胞分布。结论:tankyrase 1 siRNA在体外实验中可沉默目的基因以及抑制Tca-8113细胞增殖。
OBJECTIVE: To inhibit the proliferation of Tca-8113 cells by transfection of siRNA that inhibits the expression of tankyrase 1 in Tca-8113 cells. Methods: The siRNA against tankyrase 1 was synthesized by in vitro transcriptional synthesis and labeled with Cy-3. The recombinant plasmid was transfected into Tca-8113 cells by lipofectamine. The transfection efficiency was evaluated by fluorescence microscopy. The effect of siRNA was detected by MTT assay and flow cytometry. The effect of siRNA on tankyrase 1 expression was evaluated by Western blot. Results: Compared with the blank control group and the liposome control group, the tankyrase 1 siRNA group could down-regulate the tankyrase 1 expression in the Tca-8113 cells. The Western blot analysis showed that the expression of tankyrase 1 was down-regulated by 1/2 , Inhibited the proliferation of Tca-8113 cells (P <0.05), and changed the cell distribution at each phase of the cell cycle. Conclusion: Tankyrase 1 siRNA can silence the target gene and inhibit the proliferation of Tca-8113 cells in vitro.