不同人源性细胞中内参基因的筛选

Screening of reference genes in different human-derived cells

摘要:

目的 为筛选在不同人源性细胞中能够稳定表达的内参基因，以期为不同组织的定量研究提供一定的参考。方法 通过实时荧光定量PCR（RT-qPCR）方法检测36种人源性细胞中β-珠蛋白（β-Globin）、甘油醛-3-磷酸脱氢酶（GAPDH）、核糖体RNA（18S rRNA）和核糖核酸酶P（Rnase P）4个候选内参基因的表达量，并利用geNorm、NormFider和BestKeeper三个软件对候选内参基因的稳定性进行评价。结果 同一种细胞中，Rnase P的表达量较高；不同细胞中，18S rRNA的表达较为稳定。结论 综合分析发现候选内参基因18S rRNA虽较为稳定，但并不是最优选择。

Abstract:

Objective To screen internal reference genes that can be stably expressed in different human-derived cells, in order to provide a certain reference for quantitative research of different tissues.Methods The Expression of β-Globin, GAPDH,18S rRNA and Rnase P four candidate reference genes in 36 types of human-derived cells was detected by real-time fluorescence quantitative PCR (RT-qPCR). The stability of four candidate reference genes was evaluated using geNorm, NormFider and BestKeeper software.Results In the same type of cell, the expression of Rnase P is relatively high; in different cells, the expression of 18S rRNA is relatively stable.Conclusion Comprehensive analysis found that although the candidate reference gene 18S rRNA is relatively stable, it is not the best choice.