沉默SEPT9基因对人肝癌MHCC97-H细胞生物学特性的影响.pdfVIP

(P ﹥0.05) 。⑥细胞划痕实验结果分析，与空白对照组和阴性对照组相比， 实验组划痕宽度明显缩小，差异有统计学意义（P ＜0.05 ），空白对照组和 阴性对照组两者间划痕宽度差异无统计学意义(P ＞0.05) 。 结论：1．针对 SEPT9 基因的RNAi 靶序列设计有效；合成的 siRNA 通过脂质体介导的方 法能高效转染 MHCC97-H 细胞，并有效抑制 MHCC97-H 细胞中相应基因 的表达。2.靶向 SEPT9 的 siRNA 能显著抑制 MHCC97-H 细胞的生长和增 殖，促进细胞凋亡。3. 靶向 SEPT9 的 siRNA 能有效抑制 MHCC97-H 细胞 迁移。 关键词：SEPT9 基因；MHCC97-H 细胞；siRNA； 细胞增殖；细胞凋亡； 细胞迁移 3 Effects of SEPT9 gene silencing on biological characteristics of MHCC97-H hepatoma cell Abstract Objectives: Design and synthesise the corresponding SEPT9 - siRNA according to the gene sequence of SEPT9. explore the specific inhibitory effect and the influence of apoptosis on the growth of hepatoma cell lines MHCC97-H , and to reveal the role of SEPT9 in the tumorigenesis and the development of hepatoma. Methods: We designed 4 interference target sequence according to the SEPT9 gene nucleotide sequences，which aimed at SEPT9 gene. The biological company built 4 siRNA expression vector of SEPT9 gene，and at the same time built any gene with the negative control plasmid of green fluorescent protein gene . Preliminary experiment selected an effective interference recombinant plasmid pGPU6 / Neo - SEPT9-1253, the target sequence is GGCAGCCCATCATGAAGTTCA. Cultivated MHCC97 -H cells. Cells that were in good condition were plated in 6-well cell culture plates (CCK 8 method to detect the cell proliferation by 96 - well cell culture plates). Set up three experimental groups.They were SEPT9-siRNA group (transfection SEPT9 - siRNA expression plasmid), negative control group (transfection negative controlled plasmid) and blank group (normally cultured MHCC97 -