参考基因
小梁网
管家基因
基因
氧化应激
基因表达
生物
甘油醛3-磷酸脱氢酶
实时聚合酶链反应
细胞生物学
青光眼
分子生物学
遗传学
生物化学
神经科学
作者
Jing Wang,Hongyan Zhou,Lixia Sun,Ben Yang,Lin Zhang,Hongfeng Shi,Yajuan Zheng
标识
DOI:10.1080/10715762.2017.1282612
摘要
Oxidative stress-induced dysfunction in trabecular meshwork (TM) cells is considered a major alteration that can lead to glaucoma. Hydrogen peroxide (H2O2) is the most widely used agent for inducing oxidation in TM cells in vitro. Quantitative real-time PCR (qPCR) is an important method for studying alterations in gene expression, and suitable (i.e. invariant) reference genes must be defined to normalize expression levels. In this study, eight common reference genes, i.e. PRS18, ACTB, B2M, GAPDH, PPIA, HPRT1, YWHAZ, and TBP, were evaluated for use in studies of H2O2-induced dysfunction in TM cells. Three established algorithms, geNorm, NormFinder, and BestKeeper, were used to analyze the reference genes. ACTB expression was least affected by H2O2 treatment in TM cells, and the combination of PPIA and HPRT1 was the most suitable gene pair for normalization. GAPDH and TBP were the most unstable genes and accordingly should be avoided in experiments with TM cells. These results provide a foundation for analyses of the mechanisms underlying glaucoma, and emphasize the importance of selecting suitable reference genes for qPCR studies.
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