Evaluation and normalization of a set of reliable reference genes for quantitative sgk-1 gene expression analysis in Caenorhabditis elegans-focused cancer research

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dc.contributor.authorOzdemir, Ozguer Ulkue
dc.contributor.authorYurt, Kubra
dc.contributor.authorPektas, Ayse Nur
dc.contributor.authorBerk, Seyda
dc.date.accessioned2024-10-26T18:03:50Z
dc.date.available2024-10-26T18:03:50Z
dc.date.issued2024
dc.departmentSivas Cumhuriyet Üniversitesi
dc.description.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.
dc.description.sponsorshipScientific and Technological Research Council of Turkiye (TUBITAK) [1919B012105851]
dc.description.sponsorshipProject support was provided by the Scientific and Technological Research Council of Turkiye (TUBITAK) (Grant No. 1919B012105851). The funders did not have any roles in the study design, the collection, analysis, and interpretation of data; in writing of the man-uscript; or in the decision to submit the manuscript for publication.
dc.identifier.doi10.1080/15257770.2024.2317413
dc.identifier.issn1525-7770
dc.identifier.issn1532-2335
dc.identifier.pmid38359339
dc.identifier.scopus2-s2.0-85185672212
dc.identifier.scopusqualityQ3
dc.identifier.urihttps://doi.org/10.1080/15257770.2024.2317413
dc.identifier.urihttps://hdl.handle.net/20.500.12418/28585
dc.identifier.wosWOS:001163232600001
dc.identifier.wosqualityN/A
dc.indekslendigikaynakWeb of Science
dc.indekslendigikaynakScopus
dc.indekslendigikaynakPubMed
dc.language.isoen
dc.publisherTaylor & Francis Inc
dc.relation.ispartofNucleosides Nucleotides & Nucleic Acids
dc.relation.publicationcategoryMakale - Uluslararası Hakemli Dergi - Kurum Öğretim Elemanı
dc.rightsinfo:eu-repo/semantics/closedAccess
dc.subjectC. elegans
dc.subjectlin-35
dc.subjectsgk-1
dc.subjectnormalization
dc.subjectRT-qPCR
dc.subjectcancer
dc.titleEvaluation and normalization of a set of reliable reference genes for quantitative sgk-1 gene expression analysis in Caenorhabditis elegans-focused cancer research
dc.typeArticle

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