肿瘤中GLI1 RNA编辑的下调及其潜在功能分析  

Down-regulation of RNA editing of GLI1 in tumors and analysis of its potential function

作　　者：张颖[1] 胡亚欧[1] 何涛[1] 侯小强[1] 汪莉[1] 张部昌[2] 王玉民[1] 

机构地区：[1]军事医学科学院生物工程研究所,北京100071 [2]安徽大学生命科学学院,合肥230601

出　　处：《军事医学》2012年第4期267-271,275,共6页Military Medical Sciences

基　　金：国家自然科学基金资助项目(30800644,31100940);国家“重大新药创制”科技重大专项资助项目(2012ZX09J12106-03B)

摘　　要：目的检测癌相关基因GLI1编码区发生的A-to-I RNA编辑,比较编辑水平在肿瘤与正常细胞系中的差异,并分析编辑事件的潜在影响,以期揭示其与癌症发生发展的关联。方法通过PCR、RT-PCR及测序的方法检测GLI1编码区DNA和RNA序列上的差异,以此来识别该区域上的A-to-I RNA编辑事件和比较肿瘤与正常细胞系中编辑位点的编辑水平差异。使用多种生物信息学工具分析编辑事件的潜在功能影响。结果在人脑、乳腺、小肠组织及多种细胞系中均检测到GLI1转录本上对应染色体12∶56150891的位置发生A-to-I RNA编辑,该编辑事件导致GLI1第701位氨基酸由精氨酸转变为谷氨酸。与相应的正常细胞系相比,肝癌细胞系及乳腺癌细胞系中该位点的编辑水平明显降低。生物信息学分析表明此编辑事件能改变编码的氨基酸,并可能破坏外显子剪接增强子及影响蛋白质高级结构。结论 GLI1编码区发生的A-to-I RNA编辑在人脑、乳腺、小肠组织及多种细胞系中普遍存在。该编辑事件在肝癌和乳腺癌细胞系中的下调提示其可能与癌症的发生发展相关。Objective To detect the A-to-I RNA editing in the coding region of cancer associated gene GLI1,compare the difference in the editing level between tumor and normal cell lines,and to analyze the potential effect of this editing event in the hope of discovering the role of the RNA editing in the occurrence and progression of cancer.Methods The identification of the A-to-I RNA editing event and the comparison of the editing level were carried out by comparing the cDNA sequences with their corresponding genomic templates based on PCR,RT-PCR and sequencing methods.The potential functional implications of RNA editing were analyzed by multiple bioinformatics tools.Results An A-to-I RNA editing site mapped to chr 12∶56150891 changed the arginine residue at position 701 of the GLI1 protein to a glutamine residue,which was detected in the human brain,breast,small intestine tissues and multiple cell lines.Compared with normal controls,the editing level of this site decreased in the hepatoma and breast cancer cell lines.By bioinformatics analysis,this editing event altered amino acid residue and was predicted to destroy putative exonic splicing enhancers（ESE） and affect putative protein tertiary structure.Conclusion This A-to-I RNA editing event in the coding region of GLI1 occurs in many diverse human tissues and cell lines.Down-regulation of RNA editing of GLI1 in hepatoma and breast cancer cell lines may be associated with the occurrence and progression of cancer.

关 键 词：GLI1 编码区 A-to-IRNA编辑 生物信息学 肿瘤 

分 类 号：R730.2[医药卫生—肿瘤]