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-Misa , Katarzyna Neubauer , Ewa Wałecka-Zacharska , Bartosz Kapturkiewicz , Małgorzata Krzystek-Korpacka
AbstractBackground. Theeffect ofbowel inflammation and cancer ontheexpression of themost prevalent internal controls: ACTB, GAPDH and B2M in whole blood isunknown, although at least GAPDH occurred tobe tightly regulated and suspected of supporting cancer growth, challenging its suitability asareference. Objectives. Toevaluate theeffect of colorectal cancer (CRC) and active inflammatory bowel disease (IBD) onthestability 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 interleukin1β (IL-1β) and CCL4. Results. HPRT1, SDHA and TBP were superior normalizers inCRC and IBD. Thehighest expression variability was noted in active IBD. B2M was significantly lower in CRC but higher in IBD. GAPDH was higher in CRC andIBD. ACTB and GAPDH corresponded with CRC advancement (ρ = 0.52 and ρ = 0.53) and with clinical activity inCrohn’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 inIBD and CRC. Conclusions. Leukocyte GAPDH, ACTB, and B2M expression isaffected bybowel inflammation and cancer, rendering them unsuitable asareference inCRC and IBD.
|Journal series||Advances in Clinical and Experimental Medicine, ISSN 1899-5276, e-ISSN 2451-2680, [1230-025X], (N/A 40 pkt)|
|Publication size in sheets||0.5|
|Keywords in English||geNorm, NormFinder, BestKeeper, whole blood transcriptome, expression stability|
|License||Journal (articles only); author's original; ; after publication|
|Score||= 40.0, 15-09-2020, ArticleFromJournal|
|Publication indicators||= 0; : 2018 = 0.648; : 2018 = 1.227 (2) - 2018=1.347 (5)|
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