Identifying Stable Reference Genes for Gene Expression in Lung Cells
Author Information
Author(s): Eva Kriegova, Arsen Arakelyan, Regina Fillerova, Jaromir Zatloukal, Frantisek Mrazek, Zdenka Navratilova, Vitezslav Kolek, Roland M du Bois, Martin Petrek
Primary Institution: Palacky University & Faculty Hospital, Olomouc, The Czech Republic
Hypothesis
To identify gene(s) with stable mRNA expression in bronchoalveolar cells irrespective of various factors.
Conclusion
PSMB2 and RPL32 are suitable reference genes for normalizing qRT-PCR in bronchoalveolar cells in sarcoidosis and other interstitial lung diseases.
Supporting Evidence
- PSMB2 and RPL32 were found to be stably expressed across various patient subgroups.
- Normalization with PSMB2/RPL32 resulted in significant differences in cytokine expression.
- Traditional reference genes like ACTB and GAPDH were unsuitable for normalization.
Takeaway
Researchers found two genes, PSMB2 and RPL32, that can help scientists measure other genes accurately in lung cells, no matter the patient's background.
Methodology
mRNA expression of ten housekeeping genes was investigated by qRT-PCR in BAL cells from 71 subjects, validated in an independent cohort of 63 sarcoidosis patients and 17 controls.
Limitations
The study primarily focused on bronchoalveolar cells from interstitial lung diseases, limiting generalizability to other conditions.
Participant Demographics
71 Caucasian subjects with various lung diseases, including 45 males and 26 females.
Statistical Information
P-Value
0.004
Statistical Significance
p<0.05
Digital Object Identifier (DOI)
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