论文部分内容阅读
目的 研究小鼠肠上皮细胞在正常及轻型肠型放射病早期差异基因表达的图谱。方法 以异硫氰酸胍一步法提取小鼠正常及 1 2Gy全身照射后 3h和 96h肠上皮细胞的总RNA ,经磁珠法分离mRNA ,采用MMLV反转酶标志获得高比活性的3 2 PcDNA探针 (0 5× 1 0 6·min-1 ·μg-1 ) ,与含癌 抑癌基因、细胞周期调节、信号传导等 1 4大类共 588种鼠源性相关基因的cDNA微阵列膜杂交。结果 绘制了小鼠轻型肠型放射病早期与肠上皮再生期表达基因和差异表达基因图谱 ,发现1 2Gy辐射诱导早期 (3h) ,主要为基因表达受到抑制 ,而再生期 (96h)基因表达明显增强 ,并有新的表达基因出现。此外见cdk抑制蛋白Ⅰ基因在辐射损伤时呈稳定高表达水平。结论 1 2Gy辐射损伤 96h时基因表达综合效应表现为肠上皮损伤修复 ,cdk抑制蛋白Ⅰ基因在辐射损伤肠上皮中的高表达 ,提示其可能在监测肠上皮辐射损伤方面具有应用前景。
Objective To study the differential expression profiles of mouse intestinal epithelial cells in early stage of normal and mild enteropathy. Methods The total RNA of intestinal epithelial cells in normal and 1 Gy 2Gy mice were extracted by guanidinium thiocyanate at 3h and 96h after irradiation. MRNA was isolated by magnetic beads method. The high specific activity of 3 2 PcDNA was obtained by MMLV reverse transcriptase (0 5 × 10 6 · min -1 · μg -1) and a total of 588 mouse-related genes including tumor suppressor gene, cell cycle regulation and signal transduction Hybridization. Results The gene expression profiles and the differentially expressed genes in early stage and intestinal epithelial stage of mice with gut enteropathy were plotted. It was found that the gene expression was inhibited mainly in the early stage (3h), while the gene expression in the stage of regeneration (96 hours) was significantly enhanced , And new expressed genes appear. In addition, see cdk inhibitor Ⅰ gene showed a stable expression of high levels of radiation injury. Conclusions The combined effect of gene expression in 2Gy radiation at 96h is the repair of intestinal epithelial injury. The high expression of cdk inhibitor Ⅰ gene in the intestinal epithelium of radiation injury suggests that it may have potential application in monitoring the injury of intestinal epithelial radiation.