敲低znf662基因对u2os细胞增殖的影响.docxVIP

摘要 　　已有报道锌指蛋白662(ZNF662)与口腔鳞状细胞癌患者的生存率显著相关,但ZNF662对细胞增殖的影响未见报道。本研究采用qRT-PCR检测ZNF662基因被siRNA干扰技术敲低后在人骨肉瘤细胞U2OS细胞中的表达，检测ZNF662基因被敲低的程度，然后采用流式细胞术检测ZNF662基因敲除对U2OS细胞增殖的影响。首先通过引物设计软件设计了三对针对ZNF662 mRNA的干扰RNA序列，通过脂质体转染体外培养的人骨肉瘤细胞U2OS；用qRT-PCR转染以检测转染效率;最后使用流式细胞术分析U2OS细胞增殖的作用。我们的实验结果表明，SiRNA-3的干扰效果是最佳的，发现我们的实验结果中G0 / G1中的U2OS细胞数量小于对照组相比的细胞数，但细胞数S期和G2 / m相对增加。最终结果显示，在U2OS细胞中敲低ZNF662基因反而促进了细胞增殖的速率，这表明ZNF662基因它可能与人骨肉瘤细胞U2OS细胞的增殖有关。 关键词：ZNF662基因；细胞增殖；siRNA；U2OS细胞 第一章 Abstract 　　It has been reported that zinc finger protein 662 (ZnF662) is significantly associated with the survival rate of oral squamous cell carcinoma patients, but the effect of ZnF662 on cell proliferation has not been reported. This study uses siRNA interference technology to knock down the expression of ZnF662 gene in U2OS cells in human osteosarcoma. The ZnF662 gene is knocked low by QRTPCR, and then the influence of row of U2OS cells after the row of ZnF662 is analyzed by QRT-PCR. . Firstly, the three-pair of interference RNA sequences for ZnF662 mRNA were designed by primer design software, and transfection was transfected with human osteosarcoma cells U2OS in vitro; transfected 48 hours, the transfection efficiency was detected by QRT-PCR; finally using flow cytometry The effect of Znf662 was subjected to proliferation of U2OS cells after knocking. Our experimental results show that the interference effect of siRNA-3 is best, found that the number of U2OS cells in G0 / G1 in our experimental results is less than the number of cells compared to the control group, but the number of cells in the S phase and G2 / M It is relatively increased. The final result shows that the rate of knocking the ZnF662 gene in U2OS cells has promoted the rate of cell proliferation, indicating that the ZnF662 gene may be related to the proliferation of human osteosarcoma cell U2OS cells. 　　Key words: Zinc finger protein gene 662 (ZNF662 gene); cell proliferation; siRNA interference; U2OS cell 第一章 绪论 1.1 细胞增殖 1.1.1细胞增殖概述 　　细胞增殖