小鼠轻型肠型放射病肠上皮细胞基因表达谱的差异  

Differentially expressed genes in intestinal epithelial cells derived from mice with mild intestinal type of radiation sickness

作　　者：胡川闽[1] 粟永萍[1] 程天民[1] 王军平[1] 刘贤华[1] 冉新泽[1] 杨学森[2] 

机构地区：[1]第三军医大学预防医学系全军复合伤研究所,重庆400038 [2]第三军医大学预防医学系劳动卫生学教研室,重庆400038

出　　处：《中华放射医学与防护杂志》2002年第5期326-329,共4页Chinese Journal of Radiological Medicine and Protection

基　　金：国家自然科学基金重点资助项目 ( 39770 2 37);国家教育部<高等学校骨干教师资助计划>资助项目

摘　　要：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 differentially expressed gene profiles by intestinal epithelial cells (IECs)derived from mice with mild intestinal type of radiation sickness. Methods\ Total RNA of IECs obtained from normal animals and mice at 3 h,96 h after 12 Gy γ ray irradiation was isolated by Chomzynski's method and mRNA was purified using magnetic bead method.High specific activity 32 P\|cDNA probes (0 5×10 6 cpm/μg) were made by a single round reverse transcription with 200 U of Moloney murine leukemia virus reverse transcriptase in the presence of CDS primer (Clontech Co).The 32 P\|cDNA probes were hybridized to the microarry membrane which contains 588 genes.After scanning the hybridization signal by gel analysis system,the differentially expressed gene profiles of the IECs derived from mice at 3 h,96 h after 12 Cy γ ray irradiation were made. Results\ The differentially expressed gene profiles showed that the γ ray induced gene expression pattern at the early stage (3 h) by IECs represented mainly inhibition of gene expression.The results of the differential gene expression mainly represented the IECs damage and resistance to radiation damage.At the IECs regeneration stage (96 h after irradiation),the differentially expressed genes mainly exhibited increase of gene expression and appearance of new gene expression.Besides,cdk inhibitor Ⅰ protein gene remained at a stably high level of expression during radiation injury. Conclusion\ The differential gene expression at the regeneration stage after 12 Gy γ ray irradiation of IECs results in epithelium recovery.Perhaps the cdk inhibitor Ⅰ protein could be used to indicate the injury induced by radiation.\;

关 键 词：小鼠 肠型放射病 肠上皮细胞 基因克隆 辐射损伤 微阵列 差异基因表达 

分 类 号：R818[医药卫生—放射医学]