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Pol. Merkur. Lek (Pol. Med. J.), 2011, XXXI/181: 024-030 Maximize

Pol. Merkur. Lek (Pol. Med. J.), 2011, XXXI/181: 024-030

Title: Evaluation of β2-microglobuline expression level in gastrointestinal tract tumors due to usefulness as a reference gene in PCR method

Authors: Skrzycki M., Czeczot H., Barańczyk-Kuźma A., Mielczarek-Puta M., Chrzanowska A., Gajewska B.

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Evaluation of β2-microglobuline expression level in gastrointestinal tract tumors due to usefulness as a reference gene in PCR method


Skrzycki M., Czeczot H., Barańczyk-Kuźma A., Mielczarek-Puta M., Chrzanowska A., Gajewska B.

Medical University of Warsaw, Poland, First Faculty of Medicine, Chair and Department of Biochemistry

Quantitative and semi-quantitative determination of gene expression by PCR plays an important role in studying of tumors initiation and progression mechanisms. Selection of appropriate reference gene is a critical factor influencing the results of gene expression analysis. One of the most commonly used reference genes in PCR is β2-microglobuline (β2-M). Recent studies showed however that expression of some common reference genes might be unstable, therefore it is necessary to verify again their usefulness.
The aim of the study
was to determine the level of β2-M mRNA in normal and tumor tissues of gastrointestinal tract due to adequate selection of reference gene in gene expression studies.
Material and methods
. Samples were taken from 253 patients operated on for gastrointestinal tract tumors: 22 with oral cavity cancer, 12 with benign and 50 with malignant liver tumors, 86 with colorectal cancer, and 83 with metachronous metastases to liver. Also 56 patients with liver cirrhosis were studied, which was treated as pretumor state. Together 309 patients were studied. RNA was isolated from tissues by Chomczynski method. The expression level of ß2-M was determined by reverse transcriptase PCR (RT-PCR) and given in terms of optical density values.
Results
. Expression of β2-M was observed in all studied tissues. There were no differences between normal and tumor tissue. The level of expression of β2-M was different due to type of studied tissue (oral cavity, liver, colon).
Conclusions
. The lack of significant differences in β2-M expression level in normal and tumor tissues indicated that β2-M can be used as reference gene in studies of gene expression in gastrointestinal tract tumors. On the other hand differences of β2-M expression level in different types of tissues point to its tissue specificity and suggest application in PCR of more than one reference gene.

Key words: expression of β2-microglobuline, gastrointestinal tract tumors, PCR, reference gene

Pol. Merk. Lek., 2011, XXXI, 181, 24