小分子干扰RNA沉默RhoGDI2基因表达对结肠癌细胞增殖侵袭的影响及其机制

发布时间：2018-05-24 02:59

  本文选题：siRNA + RhoGDI　； 参考：《中国免疫学杂志》2017年04期

【摘要】：目的:应用小分子干扰RNA(siRNA)沉默RhoGDI2基因的表达,初步探讨其对结肠癌细胞增殖、迁移能力的影响及可能的机制。方法:分别运用Western blot和RT-q PCR检测结肠癌细胞株RKO、HT29、SW620、SW480、HCT116基因RhoGDI2的表达情况。设计并合成RhoGDI2 siRNA干扰序列,按照LipofectamineTM2000转染方法将siRNA干扰序列转染到目的细胞,设置实验干扰组、空白对照和阴性对照组;CCK-8实验检测细胞增殖能力,细胞划痕试验和Transwell实验检测细胞迁移、侵袭能力。结果:人结肠癌细胞株RhoGDI2表达量由高到低依次是RKO、HT29、SW620、SW480、HCT116;RKO细胞siRNA干扰后RhoGDI2表达抑制率大于70%:实验干扰组、阴性对照组、空白对照组细胞增殖率分别是(0.683±0.013)、(0.866±0.088)、(0.905±0.008),P0.05;实验干扰组细胞迁移、侵袭速率较对照组均减慢。沉默RhoGDI2基因的表达,实验组细胞E-Cadherin较对照组表达量高,Vimentin蛋白表达下降。结论:结肠癌RhoGDI2基因沉默可能通过抑制EMT进程阻止肿瘤恶性生物学行为。
[Abstract]:Aim: to silence the expression of RhoGDI2 gene by small interfering RNAs (siRNAs), and to explore the possible mechanism of its effect on the proliferation and migration of colon cancer cells. Methods: Western blot and RT-q PCR were used to detect the RhoGDI2 expression of HCT116 gene. The RhoGDI2 siRNA interference sequence was designed and synthesized, and the siRNA interference sequence was transfected into the target cell according to the LipofectamineTM2000 transfection method. The experimental interference group, the blank control group and the negative control group were used to detect the proliferation ability of the cells. Cell scratch test and Transwell assay were used to detect cell migration and invasion. Results: the order of RhoGDI2 expression in human colon cancer cell line was RKOHT29, SW620, SW480, HCT116, RKO cells, the inhibition rate of RhoGDI2 expression was higher than 70: the proliferative rate of experimental interference group, negative control group and blank control group was 0. 683 卤0. 083 卤0. 0888 卤0. 905 卤0. 008P0.05. the cell migration of experimental interference group, negative control group and blank control group were 0. 683 卤0. 088 卤0. 905 卤0. 008P0. 05 respectively. The invasion rate was slower than that of the control group. Silencing the expression of RhoGDI2 gene, the expression of Vimentin protein in the experimental group was significantly lower than that in the control group. Conclusion: RhoGDI2 gene silencing in colon cancer may prevent malignant biological behavior by inhibiting EMT progression.
【作者单位】： 吉林大学第二医院肝胆胰内科;第二军医大学长海医院检验科;上海交通大学医学院附属同仁医院肿瘤科;
【基金】：吉林省科技厅科研基金课题(2013c026-1)
【分类号】：R735.35

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