Evaluation and normalization of a set of reliable reference genes for quantitative <italic>sgk-1</italic> gene expression analysis in <italic>Caenorhabditis elegans</italic>-focused cancer research.
In: Nucleosides, Nucleotides & Nucleic Acids, 2024-02-10, S. 1-20
Online
academicJournal
AbstractMultiple signaling pathways have been discovered to play a role in aging and longevity, including the insulin/IGF-1 signaling system, AMPK pathway, TOR signaling, JNK pathway, and germline signaling. Mammalian serum and glucocorticoid-inducible kinase 1 ( sgk-1 ), which has been associated with various disorders including hypertension, obesity, and tumor growth, limits survival in C. elegans by reducing DAF-16/FoxO activity while suppressing FoxO3 activity in human cell culture. C. elegans provides significant protection for a number of genes associated with human cancer. The best known of these are the lin-35/pRb (mammalian ortholog pRb ) and CEP-1 (mammalian ortholog p53 ) genes. Therefore, in this study, we aimed to investigate the expression analyzes of sgk-1 , which is overexpressed in many types of mammalian cancer, in mutant lin-35 and to demonstrate the validation of reference genes in wild-type N2 and mutant lin-35 for C. elegans -focused cancer research. To develop functional genomic studies in C. elegans , we evaluated the expression stability of five candidate reference genes ( act-1, ama-1, cdc-42, pmp-3 , iscu-1 ) by quantitative real-time PCR using five algorithms (geNorm, NormFinder, Delta Ct method, BestKeeper, RefFinder) in N2 and lin-35 worms. According to our findings, act-1 and cdc-42 were effective in accurately normalizing the levels of gene expression in N2 and lin-35. act-1 and cdc-42 also displayed the most consistent expression patterns, therefore they were utilized to standardize expression level of sgk-1 . Furthermore, our results clearly showed that sgk-1 was upregulated in lin-35 worms compared to N2 worms. Our results highlight the importance of definitive validation using mostly expressed reference genes. [ABSTRACT FROM AUTHOR]
Titel: |
Evaluation and normalization of a set of reliable reference genes for quantitative <italic>sgk-1</italic> gene expression analysis in <italic>Caenorhabditis elegans</italic>-focused cancer research.
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Autor/in / Beteiligte Person: | Özdemir, Özgür Ülkü ; Yurt, Kübra ; Pektaş, Ayşe Nur ; Berk, Şeyda |
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Zeitschrift: | Nucleosides, Nucleotides & Nucleic Acids, 2024-02-10, S. 1-20 |
Veröffentlichung: | 2024 |
Medientyp: | academicJournal |
ISSN: | 1525-7770 (print) |
DOI: | 10.1080/15257770.2024.2317413 |
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