The ability of tumor to infiltrate the surrounding tissue is one of the main characteristics of a malignancy. Degradation the collagen type IV, a major component of the basement membrane, is an essential step in the metastasis to lymph nodes and distant organs. Matrix metalloproteinase 2 (MMP-2) and matrix metalloproteinase 9 (MMP-9) are proteolytic enzymes that digest collagen IV and other components of extracellular matrix. Tumor cells and migrated leucocytes are source of MMPs.

The aim of the study was investigation the transcriptive activity of MMP-2 and MMP-9 genes in squamous cell carcinoma of the larynx and adjacent nonneoplasmatic tissue from incision lines. Comparison of the mRNAs levels of matrix metalloproteinases in tumor samples with lymph node metastasis and without metastatic lymph node was made as well.

The study included 32 patients with primary LSCC in histological grade G1 – G3, who underwent surgical treatment at the E.N.T. Clinics of Medical University of Silesia in Katowice. In examined group there were 13 patients with lymph node metastasis (N³1) and 19 patients without metastatic lymph node.

Quantification of genes expression was performed by real time QRT-PCR technique with a SYBR Green I chemistry using an Opticon™ DNA Engine Sequence Detector (MJ Research, USA). Oligonucleotide primers specific for mRNAs MMP-2 and MMP-9 were designed on the basis of reference sequences (GeneBank accession no. NM_004530 and NM_004994 respectively) by using Primer Express äv.1.0 ABI PRISM (Applied Biossystems). The obtained results were recalculated per mg total RNA.

We have detected mRNA of MMP-2 and MMP-9 in all samples. In tumor center expression of MMP-2 gene was 4-fold higher than MMP-9, whereas in normal tissues was 40-fold higher. Moreover concentration of mRNA MMP-2 (Me=5,2´10⁴ copies/mgRNA) was higher in tumors compared with adjacent normal tissues (Me=1,6´10⁴ copies/mgRNA) (p=0,001; Wilcoxon test). The transcriptive activity of MMP-9 in cancer area (Me=1,2´10⁴ copies/mgRNA) was significantly higher than in surrounding nonneoplasmatic tissues (Me=4,0´10² copies/mgRNA) (p<0,001)

In both groups LSCC with and without lymph node metastasis expression of MMP-2 and MMP-9 were in the same levels (NS, U Mann-Whitney test).Additionally, we determined high positive correlation between the expression of MMP-2 and MMP-9 genes in tumor tissues (R=0,74; p=0,0000; Spearman,s rank correlation test).

In summary, a higher expression of matrix metalloproteinases in tumor is connected with its larger susceptibility to invasion and metastasis.

Increased transcriptive activity of MMP-2 gene is associated with increased transcriptive activity of MMP-9 gene.

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