ACTB and SDHA Are Suitable Endogenous Reference Genes for Gene Expression Studies in Human Astrocytomas Using...

Area of Science:

• Oncology
• Molecular Biology
• Neuroscience

Background:

• Quantitative real-time reverse-transcription polymerase chain reaction (qRT-PCR) is crucial in experimental oncology.
• Existing reference gene studies for human glioma are limited, focusing only on specific comparisons.
• A need exists for universally applicable reference genes for glioma research, including pre- and post-treatment analyses.

Purpose of the Study:

• To validate a broader range of reference genes for human glioma research.
• To identify stable endogenous control genes suitable for various glioma subtypes and treatment conditions.
• To enable more reliable gene expression analysis in glioblastoma studies.

Main Methods:

• A panel of six candidate reference genes (ACTB, GAPDH, POLR2A, RPL13A, SDHA, TBP) representing different physiological pathways were tested.
• Gene expression stability was analyzed across common glioma groups (WHO grades II-IV), including primary, secondary, and recurrent glioblastomas, and samples before and after radiotherapy/chemotherapy.
• Longitudinal expression stability was assessed in eight individual patients.

Main Results:

• ACTB, GAPDH, and RPL13A exhibited higher expression levels compared to SDHA, POLR2A, and TBP.
• ACTB, GAPDH, and RPL13A showed differential expression between astrocytoma grade II and primary glioblastoma.
• SDHA and ACTB were identified as the most stable reference genes using NormFinder software, with SDHA showing the lowest intragroup variation.

Conclusions:

• ACTB and SDHA are recommended as the most stable reference genes for data normalization in human glioma gene expression studies.
• These genes are suitable for various treatment regimens and glioma grades (WHO II-IV).
• The findings support the use of ACTB and SDHA for reliable gene expression analysis in diverse glioblastoma research settings.