Медицинская библиотека · Gene · 2025

Reference genes for quantitative real-time polymerase chain reaction in in vitro non-alcoholic fatty liver disease.

Chen Jingzhi, Chen Xingmi, Liu Ying, Lyu Chunming, Xu Ming, Yang Yang

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Аннотация

BACKGROUND

In metabolic dysfunction-associated fatty liver disease (MAFLD) research, reference genes for qPCR are crucial but often unvalidated.

METHODS

RNA-seq was performed in control and free fatty acid (FFA) treated AML12 cells, and the candidates for reference gene were selected by previous literatures and filtered by dual-index ranking via normalization of coefficient variation and Log2FoldChange in the RNA-seq library. qPCR data was further analyzed via NormFinder, BestKeeper, geNorm, ΔCt and RefFinder to assess reference gene expression stability.

RESULTS

FFA treated cells showed a significantly increased lipid droplet accumulation. RNA-seq dual-index ranking and RefFinder identified that Ywhaz was the most stable reference gene.

CONCLUSION

Our finding revealed that Ywhaz is the most stable reference gene for qPCR in the AML12 cell model of FFA-induced MAFLD, and provided a reliable procedure for screening reference genes.

Для цитирования: Chen Jingzhi, Chen Xingmi, Liu Ying, Lyu Chunming, Xu Ming, Yang Yang. Reference genes for quantitative real-time polymerase chain reaction in in vitro non-alcoholic fatty liver disease.. Gene. 2025. DOI: 10.1016/j.gene.2025.149931.

Предметные рубрики MeSH

Mice 14-3-3 Proteins Cell Line Fatty Acids, Nonesterified Gene Expression Profiling Non-alcoholic Fatty Liver Disease Real-Time Polymerase Chain Reaction Reference Standards Animals

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