西安交通大学学报（医学版）2017，Vol.38Issue(4)：497-501,535,6.DOI:10.7652/jdyxb201704006
U6和let-7a作为定量检测miRNAs的内参在大鼠软骨中稳定性的比较
Comparison between Let-7a and U6 as an internal reference for RT-qPCR of miRNAs in rat cartilage
摘要
Abstract
Objective To evaluate the stability of U6 and let-7a as internal reference genes of miRNAs in RTqPCR by using femoral head samples of cartilage tissue from inbred DA rats.Methods Total RNA was extracted from femoral head cartilage tissues of female DA rats at three different time points,i.e.at birth (D0),ablactation (D21) and maturation (D42).The expressions of different miRNAs (miR-1,-25,-26a,-140,-146a,-150,-181a,-195,-223 and-337) were detected by RT-qPCR using U6 or let-7a as the internal reference.The two sets of miR expression were compared with the results from Solexa sequencing in our pioneer work to evaluate the stability of the two internal references.Results The relative values of U6 (P =0.045) and let-7a (P =0.021 5) revealed significant difference in the D42 sample.Both in U6 and let-7a systems,miR-26a,-140,-223,and-337 showed a similar tendency in expression and quantification but miR-1 and-146a did not have significant differences.miR-25,-150,-181a and-195 differed significantly (P＜0.05).Comparison of absolute quantification results between the two generations' sequencing showed that let-7a is more stable than U6.Conclusion Let-7a is more suitable to be used as the internal reference gene in RT-qPCR for miRNAs in cartilage tissue.关键词
软骨/内参/RT-qPCR/U6/Let-7a/microRNAs(miRNAs)Key words
cartilage/internal reference/RT-qPCR/U6/Let-7a/microRNAs (miRNAs)分类
医药卫生引用本文复制引用
易林,郭华,郭东贤,闵自信,袁莹,赵益童,韩燕,钟楠楠,孙健..U6和let-7a作为定量检测miRNAs的内参在大鼠软骨中稳定性的比较[J].西安交通大学学报（医学版）,2017,38(4):497-501,535,6.基金项目
国家自然科学基金资助项目(No.81371986,81301598) （No.81371986,81301598）
中国博士后科学基金(No.2014T70928,2013M530427) （No.2014T70928,2013M530427）
中央高校基本研究基金(No.xjj2015073) Supported by the National Natural Science Foundation of China (No.81371986 and 81301598),China Postdoctoral Science Foundation (No.2014T70928 and 2013M530427),and the Fundamental Research Funds for the Central Universities (No.xjj2015073) （No.xjj2015073）
