Open Access

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

  • Published online on: September 17, 2021     https://doi.org/10.3892/ol.2021.13052
  • Article Number: 791
  • Copyright: © Ahn et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

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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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November-2021
Volume 22 Issue 5

Print ISSN: 1792-1074
Online ISSN:1792-1082

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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