靶向干扰PROK1基因对结直肠癌细胞HT29增殖和迁移的影响

发布时间：2018-02-17 02:36

本文关键词： PROK RNA干扰结直肠癌细胞增殖迁移　出处：《基因组学与应用生物学》2017年04期 　论文类型：期刊论文

【摘要】：为研究前动力蛋白1(PROK1)对结直肠癌细胞增殖与迁移的作用机制,设计合成3条针对PROK1基因的靶向干扰序列,采用荧光显微镜观察干扰片段的转染情况,实时荧光定量PCR分析靶向干扰PROK1基因后PROK1 mRNA下调的表达水平,采用MTT、平板克隆形成试验、transwell迁移实验检测敲除PROK1后对HT29细胞增殖和迁移的影响。结果显示:荧光标记的Cy3-siRNA成功转染到HT29细胞中;3条靶向PROK1基因siRNA-62、siRNA-341、siRNA-1121转染HT29细胞48 h后,沉默效率分别为35%、70%、50%;与阴性对照组相比,siRNA-341组细胞增殖明显被抑制,24 h抑制最显著(p0.01);siRNA-341组的细胞克隆形成率(19.3±1.989 9)与空脂质体对照组(30.3±3.780 6)和阴性对照组(32.3±2.404 1)相比显著减少(p0.05);siRNA-341组通过小室迁移的细胞数(76±9.539 3)与空脂质体对照组(212±2.645 7)和阴性对照组(193±7.539 8)相比明显减少(p0.01)。本研究发现PROK1基因促进结直肠癌细胞的增殖和转移,PROK1基因有望成为治疗结直肠癌潜在的靶位点。
[Abstract]:In order to study the mechanism of Prok1) on the proliferation and migration of colorectal cancer cells, three targeted interference sequences targeting PROK1 gene were designed and synthesized. The transfection of interference fragments was observed by fluorescence microscope. Real-time fluorescence quantitative PCR was used to analyze the down-regulation of PROK1 mRNA expression after targeted interference with PROK1 gene. The effects of PROK1 knockout on the proliferation and migration of HT29 cells were detected by using MTT and plate clone formation assay. The results showed that three PROK1 targeting siRNA-62siRNA-341siRNA-1121 were successfully transfected into HT29 cells for 48 h. Compared with the negative control group, the cell proliferation of the siRNA-341 group was significantly inhibited for 24 h, and the cell clone formation rate was significantly decreased in the p0.01 siRNA-341 group (19.3 卤1.989 9) compared with the empty liposome control group (30.3 卤3.786) and the negative control group (32.3 卤2.404 1). Compared with empty liposome control group (212 卤2.645 7) and negative control group (193 卤7.539 8), the number of cells migrating through the cell of p0.05siRNA-341 group decreased significantly (p 0.01). It was found that PROK1 gene could promote the proliferation and metastasis of colorectal cancer cell line PROK1 gene. A potential target for colorectal cancer.
【作者单位】：中南大学湘雅医学院医学检验系;湖南省肿瘤医院暨中南大学湘雅医学院附属肿瘤医院;
【基金】：湖南省长沙市科技计划项目(k1501016-31) 长沙市科技项目(K1403158-61) 中南大学国家级大学生创新类项目(201610533027)共同资助
【分类号】：R735.34

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