{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,8,18]],"date-time":"2026-08-18T04:09:46Z","timestamp":1787026186665,"version":"build-2736575974"},"reference-count":38,"publisher":"Frontiers Media SA","license":[{"start":{"date-parts":[[2025,12,4]],"date-time":"2025-12-04T00:00:00Z","timestamp":1764806400000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/100031478","name":"NextGenerationEU","doi-asserted-by":"publisher","award":["Program Contract of 8 December 2023, Class: 643-02\/23-01\/00016, Reg. no. 533-03-23-0006"],"award-info":[{"award-number":["Program Contract of 8 December 2023, Class: 643-02\/23-01\/00016, Reg. no. 533-03-23-0006"]}],"id":[{"id":"10.13039\/100031478","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["frontierspartnerships.org"],"crossmark-restriction":true},"short-container-title":["Br. J. Biomed. Sci."],"abstract":"<jats:p>The gold standard for assessing expression of miRNAs, small molecules involved in numerous biological processes, is reverse transcription-quantitative polymerase chain reaction (RT-qPCR). The reliability of RT-qPCR analysis results largely depends on accurate data normalization and the selection of an appropriate reference gene. This study evaluated the stability of five candidate reference genes\u2014miR-525, miR-520c, SNORD48, miR-135b, and miR-143\u2014in human placental samples. GeNorm, NormFinder, BestKeeper, and the delta Ct-method were used to evaluate gene expression stability. The effect of reference gene selection for normalization of target miRNAs (miR-1537, miR-190b, miR-16, miR-21, and miR-146a) expression in term placental samples from smokers and non-smokers was also investigated. All statistical tools identified miR-525, miR-520c, and SNORD48 as the three most stable reference genes, except for GeNorm, which recommends the combination of the first two genes. Normalization using SNORD48 and miR-525 produced comparable results for miR-21 expression in the placental samples, both in smokers and non-smokers, whereas normalization with miR-143 yielded markedly different outcomes compared to SNORD48 and miR-525. These findings highlight the considerable impact of reference gene selection on RT-qPCR results, emphasizing the importance of careful validation to avoid misinterpretation of gene expression data.<\/jats:p>","DOI":"10.3389\/bjbs.2025.15354","type":"journal-article","created":{"date-parts":[[2025,12,4]],"date-time":"2025-12-04T06:10:30Z","timestamp":1764828630000},"update-policy":"https:\/\/doi.org\/10.3389\/crossmark-policy","source":"Crossref","is-referenced-by-count":2,"title":["Influence of Reference Gene Selection on miRNA Quantification by RT-qPCR in Human Placental 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