Reference genes identified in SH-SY5Y cells using custom-made gene arrays with validation by quantitative polymerase...

Area of Science:

• Neuroscience
• Molecular Biology
• Genetics

Background:

• Transcriptomic methods are crucial for understanding physiological and pathological processes.
• Accurate gene expression analysis relies on stable reference genes for normalization.
• Housekeeping genes often exhibit variability across tissues and experimental conditions.

Purpose of the Study:

• To identify reliable reference genes for normalizing gene expression in a cellular model of Alzheimer's disease.
• To validate reference gene stability in a human tau-expressing SH-SY5Y neuroblastoma cell line treated with amyloid-beta (Abeta(42)) peptide.
• To ensure accurate transcriptomic analysis in Alzheimer's disease research.

Main Methods:

• Development of a custom microarray based on public database gene candidates.
• Quantitative real-time polymerase chain reaction (qRT-PCR) for candidate gene analysis.
• Analysis of gene stability using geNorm software.

Main Results:

• GAPD, M-RIP, and POLR2F were identified as stable reference genes.
• These genes demonstrated reliability across different differentiation states of the SH-SY5Y cells.
• Reference gene stability was confirmed irrespective of Abeta(42) peptide treatment.

Conclusions:

• GAPD, M-RIP, and POLR2F are validated as suitable reference genes for Alzheimer's disease research in SH-SY5Y cells.
• The identified genes provide a reliable basis for accurate transcriptomic normalization in this specific cellular model.
• This finding supports robust mechanistic insights into Alzheimer's disease pathogenesis through gene expression studies.