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HMBS is the most suitable reference gene for RT‑qPCR in human HCC tissues and blood samples

  • Authors:
    • Hye Ri Ahn
    • Geum Ok Baek
    • Moon Gyeong Yoon
    • Juan A Son
    • Donglim You
    • Jung Hwan Yoon
    • Hyoung Jung Cho
    • Soon Sun Kim
    • Jae Yeon Cheong
    • Jung Woo Eun
  • View Affiliations / Copyright

    Affiliations: Department of Gastroenterology, Ajou University School of Medicine, Suwon 16499, Republic of Korea, Department of Pathology, College of Medicine, The Catholic University of Korea, Seoul 06591, Republic of Korea
    Copyright: © Ahn et al. This is an open access article distributed under the terms of Creative Commons Attribution License.
  • Article Number: 791
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    Published online on: September 17, 2021
       https://doi.org/10.3892/ol.2021.13052
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Abstract

Reverse transcription‑quantitative (RT‑q) PCR is the most feasible and useful technique for identifying and evaluating cancer biomarkers; however, the method requires suitable reference genes for gene expression analysis. The aim of the present study was to identify the most suitable reference gene for the normalization of relative gene expression in human hepatocellular carcinoma (HCC) tissue and blood samples. First, 14 candidate reference genes were selected through a systematic literature search. The expression levels of these genes (ACTB, B2M, GAPDH, GUSB, HMBS, HPRT1, PGK1, PPIA, RPLP0, RPL13A, SDHA, TBP, TFRC and YWHAZ) were evaluated using human multistage HCC transcriptome data (dataset GSE114564), which included normal liver (n=15), chronic hepatitis (n=20), liver cirrhosis (n=10), and early (n=18) and advanced HCC (n=45). From the 14 selected genes, five genes, whose expression levels were stable in all liver disease statuses (ACTB, GAPDH, HMBS, PPIA and RPLP0), were further assessed using RT‑qPCR in 40 tissues (20 paired healthy tissues and 20 tissues from patients with HCC) and 40 blood samples (20 healthy controls and 20 samples from patients with HCC). BestKeeper statistical algorithms were used to identify the most stable reference genes, of which HMBS was found to be the most stable in both HCC tissues and blood samples. Therefore, the results of the present study suggest HMBS as a promising reference gene for the normalization of relative RT‑qPCR techniques in HCC‑related studies.
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Copy and paste a formatted citation
Spandidos Publications style
Ahn HR, Baek GO, Yoon MG, Son JA, You D, Yoon JH, Cho HJ, Kim SS, Cheong JY, Eun JW, Eun JW, et al: <em>HMBS</em> is the most suitable reference gene for RT‑qPCR in human HCC tissues and blood samples. Oncol Lett 22: 791, 2021.
APA
Ahn, H.R., Baek, G.O., Yoon, M.G., Son, J.A., You, D., Yoon, J.H. ... Eun, J.W. (2021). <em>HMBS</em> is the most suitable reference gene for RT‑qPCR in human HCC tissues and blood samples. Oncology Letters, 22, 791. https://doi.org/10.3892/ol.2021.13052
MLA
Ahn, H. R., Baek, G. O., Yoon, M. G., Son, J. A., You, D., Yoon, J. H., Cho, H. J., Kim, S. S., Cheong, J. Y., Eun, J. W."<em>HMBS</em> is the most suitable reference gene for RT‑qPCR in human HCC tissues and blood samples". Oncology Letters 22.5 (2021): 791.
Chicago
Ahn, H. R., Baek, G. O., Yoon, M. G., Son, J. A., You, D., Yoon, J. H., Cho, H. J., Kim, S. S., Cheong, J. Y., Eun, J. W."<em>HMBS</em> is the most suitable reference gene for RT‑qPCR in human HCC tissues and blood samples". Oncology Letters 22, no. 5 (2021): 791. https://doi.org/10.3892/ol.2021.13052
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x
Spandidos Publications style
Ahn HR, Baek GO, Yoon MG, Son JA, You D, Yoon JH, Cho HJ, Kim SS, Cheong JY, Eun JW, Eun JW, et al: <em>HMBS</em> is the most suitable reference gene for RT‑qPCR in human HCC tissues and blood samples. Oncol Lett 22: 791, 2021.
APA
Ahn, H.R., Baek, G.O., Yoon, M.G., Son, J.A., You, D., Yoon, J.H. ... Eun, J.W. (2021). <em>HMBS</em> is the most suitable reference gene for RT‑qPCR in human HCC tissues and blood samples. Oncology Letters, 22, 791. https://doi.org/10.3892/ol.2021.13052
MLA
Ahn, H. R., Baek, G. O., Yoon, M. G., Son, J. A., You, D., Yoon, J. H., Cho, H. J., Kim, S. S., Cheong, J. Y., Eun, J. W."<em>HMBS</em> is the most suitable reference gene for RT‑qPCR in human HCC tissues and blood samples". Oncology Letters 22.5 (2021): 791.
Chicago
Ahn, H. R., Baek, G. O., Yoon, M. G., Son, J. A., You, D., Yoon, J. H., Cho, H. J., Kim, S. S., Cheong, J. Y., Eun, J. W."<em>HMBS</em> is the most suitable reference gene for RT‑qPCR in human HCC tissues and blood samples". Oncology Letters 22, no. 5 (2021): 791. https://doi.org/10.3892/ol.2021.13052
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