期刊
SCIENTIFIC REPORTS
卷 11, 期 1, 页码 -出版社
NATURE PORTFOLIO
DOI: 10.1038/s41598-021-98869-x
关键词
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资金
- National Research, Development and Innovation Office of Hungary [K119493, K135231, K128369, NKP-2018-1.2.1-NKP-2018-00005]
- BME-Biotechnology FIKP Grant of EMMI (BME FIKP-BIO)
- Hungarian Thematic Excellence Programme [TKP2020-NKA-26]
This study aimed to identify stable reference genes with low expression variability among different cell lines, leading to the proposal of IPO8, PUM1, HNRNPL, SNW1, and CNOT4 as suitable for gene expression normalization. CNOT4 was also found to be the most stable gene even under serum starvation conditions.
Reverse transcription-quantitative real-time PCR (RT-qPCR) is a ubiquitously used method in biological research, however, finding appropriate reference genes for normalization is challenging. We aimed to identify genes characterized with low expression variability among human cancer and normal cell lines. For this purpose, we investigated the expression of 12 candidate reference genes in 13 widely used human cancer cell lines (HeLa, MCF-7, A-549, K-562, HL-60(TB), HT-29, MDA-MB-231, HCT 116, U-937, SH-SY5Y, U-251MG, MOLT-4 and RPMI-8226) and, in addition, 7 normal cell lines (HEK293, MRC-5, HUVEC/TERT2, HMEC, HFF-1, HUES 9, XCL-1). In our set of genes, we included SNW1 and CNOT4 as novel candidate reference genes based on the RNA HPA cell line gene data from The Human Protein Atlas. HNRNPL and PCBP1 were also included along with the classical reference genes ACTB, GAPDH, IPO8, PPIA, PUM1, RPL30, TBP and UBC. Results were evaluated using GeNorm, NormFiner, BestKeeper and the Comparative Delta Ct methods. In conclusion, we propose IPO8, PUM1, HNRNPL, SNW1 and CNOT4 as stable reference genes for comparing gene expression between different cell lines. CNOT4 was also the most stable gene upon serum starvation.
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