siRNA干扰slug基因表达对IEC-6细胞放射线敏感性影响

发布时间：2018-05-16 04:17

本文选题：slug基因 + RNA干扰　；参考：《青岛大学医学院学报》2017年01期

【摘要】：目的探讨siRNA干扰slug基因表达对体外肠隐窝上皮细胞IEC-6接受X线照射的敏感性影响及其机制。方法以体外培养大鼠小肠隐窝上皮细胞IEC-6为研究对象,将转染siRNA-slug(实验组)、转染siRNA(阴性对照组)和未转染siRNA(空白对照组)的细胞进行X射线照射处理,24h后,用Western blot法检测各组细胞的Slug和细胞P53正向凋亡上调因子(PUMA)蛋白的表达,CCK8法检测细胞在不同剂量X射线照射下的生长抑制情况,流式细胞仪检测细胞凋亡情况。结果实验组Slug蛋白表达与阴性对照组和空白对照组比较明显降低,PUMA蛋白表达明显高于阴性对照组和空白对照组,差异有显著性(F=1 760、421,P0.01)。与阴性对照组和空白对照组比较,实验组细胞的凋亡率增加,差异有统计学意义(F=33.02,P0.01)。在0~6Gy剂量范围内,随着X线照射剂量的增加,各组细胞的增殖活性均降低(F=6.595~31.470,P0.05);在6Gy剂量X线照射下,实验组细胞生长抑制率、早期+晚期凋亡率较阴性对照组和空白对照组明显增高,差异有统计学意义(F=47.700、33.020,P0.01)。结论 siRNA干扰slug基因的表达能够使促凋亡的PUMA蛋白表达增加,从而增强IEC-6细胞对X射线的敏感性。
[Abstract]:Objective to investigate the effect of siRNA interfering slug gene expression on the sensitivity and mechanism of X ray irradiation to IEC-6 of intestinal crypt epithelial cells in vitro. Methods IEC-6 of rat intestinal crypt epithelial cells was cultured in vitro. The cells transfected with siRNA-slug (experimental group, negative control group) and untransfected siRNAs (blank control group) were treated with X ray irradiation for 24 hours. Western blot assay was used to detect the expression of Slug and p53 positive apoptosis up-regulation factor (Puma) protein. CCK8 method was used to detect the growth inhibition of cells exposed to different doses of X-rays, and flow cytometry was used to detect the apoptosis of cells. Results compared with the negative control group and the blank control group, the expression of Slug protein in the experimental group was significantly lower than that in the negative control group and the blank control group, and the difference was significant (P 0.01). Compared with the negative control group and the blank control group, the apoptosis rate of the experimental group increased, and the difference was statistically significant. In the range of 0~6Gy dose, the proliferative activity of each group decreased with the increase of X-ray irradiation dose. The rate of late apoptosis in early stage was significantly higher than that in negative control group and blank control group, and the difference was statistically significant (P 0.01). Conclusion siRNA interference with the expression of slug gene can increase the expression of apoptotic PUMA protein and enhance the sensitivity of IEC-6 cells to X-ray.
【作者单位】：青岛大学附属医院麻醉科;青岛大学附属医院外科;青岛大学附属医院小儿外科;
【基金】：山东省自然科学基金资助项目(2014ZRB01198)
【分类号】：R730.55

【相似文献】