miR-138靶基因hTERT的生物信息学预测及鉴定  

Prediction and identification of target gene hTERT of miR-138 with bioinformatics technique

作　　者：陈陵[1] 陈先华[2] 梁光萍[2] 范亚川[1] 王国威[1] 宋航[1] 邹利全[1] 

机构地区：[1]中国人民解放军第三二四医院消化内科,重庆400020 [2]中国人民解放军第三军医大学西南医院烧伤研究所,重庆400038

出　　处：《实用肿瘤杂志》2013年第6期579-582,共4页Journal of Practical Oncology

基　　金：国家自然科学基金(30900679);重庆市自然科学基金资助(cstc2011jjA10111;cstc2011jjA10114;cstc2012 jjA10145)

摘　　要：目的预测并鉴定miR-138的靶基因人端粒酶反转录酶(human telomerase reverse transcriptase,hTERT)。方法采用生物信息学技术,预测miR-138与hTERT 3'UTR结合位点及结合后稳定性。分别采用RTqPCR、Western blot技术检测过表达miR-138后MKN-45细胞内hTERT在mRNA与蛋白水平的变化。采用双荧光素酶实验及突变实验,验证miR-138与hTERT mRNA的结合位点。结果生物信息学技术预测显示,miR-138可稳定结合于hTERT 3'UTR。miR-138过表达后可明显抑制MKN-45细胞内hTERT蛋白表达水平(P<0.01),但对hTERT mRNA水平无显著影响(P>0.05)。双荧光素酶实验证实miR-138可稳定结合于hTERT 3'UTR相应位点。结论 miR-138通过直接结合于靶基因hTERT 3'UTR抑制hTERT蛋白表达。miR-138可能成为基于miRNA的肿瘤基因治疗的新靶标。Objective To predict and identify the target gene human telomerase reverse transcriptase （hTERT） of miR-138. Methods The bioinformatics analysis was employed to predict miR-138 binding sites with hTERT 3＇UTR. RT-qPCR and Western-blot were used to examine the inhibitory effect on hTERT in human gastric carcinoma MKN-45 cells by miR-138. Furthermore, the binding site of miR-138 to hTERT 3＇UTR was examined using dual lueiferase reporter assay. Results The bioinfomlatics analysis showed stable binding site of miR-138 and hTERT 3＇UTR. Over-expression of miR-138 in MKN-45 cells significantly suppressed the expression of hTERT protein（ P 〈 0.01 ）, but not hTERT mRNA （ P 〉 0.05 ）. The dual luciferase reporter assay verified the binding site of miR-138 to hTERT 3＇UTR. Conclusion miR-138 can stably bind to hTERT 3＇UTR and suppress the hTERT expression,which makes miR-138 a promising target in cancer gene therapy.

关 键 词：胃肿瘤 微RNAS 端粒 RNA指导的DNA聚合酶 萤光素酶类 逆转录聚合酶链反应 基因表达 

分 类 号：R730.2[医药卫生—肿瘤]