负载miR-138复制缺陷型腺病毒对hTERT表达的影响  

Effects of replication-deficient adenovirus carrying miR-138 on the hTERT expression

作　　者：刘志鹏[1] 王宗华[2] 梁光萍[3] 熊晓峰[1] 范亚川[1] 邹利全[1] 王洪斌[1] 陈陵[1] 李学成[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]第三军医大学护理学院,重庆400038 [3]第三军医大学西南医院烧伤研究所,重庆400038

出　　处：《重庆医学》2011年第29期2947-2948,2951,共3页Chongqing medicine

基　　金：重庆市自然科学基金资助(CSTC2008BB5274;CSTC2009BB5314;CSTC2010BB5158)

摘　　要：目的探讨以复制缺陷型腺病毒为载体,上调人端粒酶逆转录酶(hTERT)调控相关miR-138表达丰度后,能否有效抑制靶基因hTERT的表达。方法 将负载hTERT调控相关miR-138的复制缺陷型腺病毒以200∶1的感染复数(MOI),感染人胃癌BGC-823细胞;采用实时荧光定量PCR(real-time PCR)检测BGC-823细胞内miR-138和hTERT mRNA的表达丰度;Western Blot检测hTERT蛋白表达。结果 与阴性对照腺病毒相比,负载miR-138的腺病毒感染BGC-823细胞后,可显著提高miR-138表达丰度(P<0.05);而未感染腺病毒的空白对照组与阴性对照组相比差异无统计学意义(P>0.05)。与感染阴性对照腺病毒的BGC-823细胞相比,感染负载人miR-138腺病毒(Ad-miR138)的BGC-823细胞内hTERT蛋白表达有所下调。结论 以复制缺陷型腺病毒为载体,可有效上调hTERT调控相关miR-138的表达丰度,进而可有效抑制miR-138靶基因hTERT的表达水平。Objective To identify effects of the replication-deficient recombinant adenovirus carrying miR-138 on the expression level miR-138 and the target gene hTERT.Methods The human gastric cancer cell line BGC-823 were infected with the replication-deficient recombinant adenovirus carrying miR-138 by the MOI of 200：1.The expression levels of miR-138 and hTERT mRNA were inuestigated by real-time PCR.The expression of hTERT protein was detected by western blot.Results Compared to the negative control,after infecting the replication-deficient recombinant adenovirus carrying miR-138,BGC-823 cells expressed higher level of miR-138（9.152±2.641 vs.0.928±0.199,P0.05）.No significant difference was observed between the ZERO group（BGC-823 infecting no adenovirus） and negative control（1.085±0.478 vs.0.928±0.199,P0.05）.The expression of hTERT protein was down-regulated in the BGC-823 cells after infected adenovirus carrying miR-138.Conclusion The replication-deficient recombinant adenovirus carrying miR-138 can up-regulate the expression of miR-138 effectively and suppress the expression of hTERT protein.

关 键 词：RNA指导的DNA聚合酶 胃肿瘤 复制缺陷型腺病毒载体 

分 类 号：R730.2[医药卫生—肿瘤]