Advances in Clinical and Experimental Medicine

Title abbreviation: Adv Clin Exp Med
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ISSN 1899–5276 (print)
ISSN 2451-2680 (online)
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Advances in Clinical and Experimental Medicine

2020, vol. 29, nr 5, May, p. 547–556

doi: 10.17219/acem/118845

Publication type: original article

Language: English

License: Creative Commons Attribution 3.0 Unported (CC BY 3.0)

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Whole blood ACTB, B2M and GAPDH expression reflects activity of inflammatory bowel disease, advancement of colorectal cancer, and correlates with circulating inflammatory and angiogenic factors: Relevance for real-time quantitative PCR

Iwona Bednarz-Misa1,A,B,C,D,F, Katarzyna Neubauer2,B,C,E,F, Ewa Zacharska3,B,E,F, Bartosz Kapturkiewicz4,B,E,F, Małgorzata Krzystek-Korpacka1,A,B,C,D,F

1 Department of Medical Biochemistry, Wroclaw Medical University, Poland

2 Department of Gastroenterology and Hepatology, Wroclaw Medical University, Poland

3 Department of Food Hygiene and Consumer Health, Wrocław University of Environmental and Life Sciences, Poland

4 First Department of Oncological Surgery of Lower Silesian Oncology Center, Wrocław, Poland


Background. The effect of bowel inflammation and cancer on the expression of the most prevalent internal controls: ACTB, GAPDH and B2M in whole blood is unknown, although at least GAPDH occurred to be tightly regulated and suspected of supporting cancer growth, challenging its suitability as a reference.
Objectives. To evaluate the effect of colorectal cancer (CRC) and active inflammatory bowel disease (IBD) on the stability of ACTB, B2M, GAPDH, HPRT1, SDHA, and TBP leukocyte expression.
Material and Methods. Gene expression in controls and CRC and IBD patients (n = 21/18/25) was evaluated in real-time quantitative polymerase chain reaction (RT-qPCR) using NormFinder, geNorm, BestKeeper, and comparative ΔCt method, and validated by comparison with absolute quantification of interleukin 1β (IL-1β) and CCL4.
Results. HPRT1, SDHA and TBP were superior normalizers in CRC and IBD. The highest expression variability was noted in active IBD. B2M was significantly lower in CRC but higher in IBD. GAPDH was higher in CRC and IBD. ACTB and GAPDH corresponded with CRC advancement (ρ = 0.52 and ρ = 0.53) and with clinical activity in Crohn’s disease (ρ = 0.44 and ρ = 0.57) and ulcerative colitis (GAPDH: ρ = 0.72). ACTB, B2M and GAPDH correlated with circulating inflammatory/angiogenic indices, differently in IBD and CRC.
Conclusion. Leukocyte GAPDH, ACTB, and B2M expression is affected by bowel inflammation and cancer, rendering them unsuitable as a reference in CRC and IBD.

Key words

geNorm, NormFinder, BestKeeper, whole blood transcriptome, expression stability

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