Matrix metalloproteinases and inhibitors of matrix metalloproteinas as markers of laryngeal cancer metastases
Authors: Kovtunenko Alexander , Tymchuk Sergey , Shportko Bogdan 
Published: July 2020

Topicality: Today, laryngeal cancer occupies an important place in the structure of tumors of the head and neck. The survival rate of patients with laryngeal cancer remains unchanged for a longtime. One of the main mechanisms of tumor growth and metastasis is invasion into the surrounding tissues, followed by the expansion of tumor cells. This process is regulated by a system of enzymes – matrix metalloproteinases (MMP) and their inhibitors – tissue inhibitors of matrix metalloproteinases (TIMP).
Aim: To study the in formativeness of immunohistochemical study and determining of the expression of molecular markers MMP-1; MMP-9; TIMR-1 and TIMR-2 in patients with laryngeal cancer, to determine the possibility of predicting regional metastases and recurrences.
Materials and methods: 70 patients with laryngeal cancer of III-IV stages (T3-4N0-3M0) and II clinical group were understudy. Of these, 39 patients from the main group had regional metastases of laryngeal cancer, in 31 patients of the comparison group regional metastases were not detected. The age of patients ranged from 33 to 74 years. The average age of patients was 61.9 years. Male patients made up an absolute majority of 99% (No=69). All patients were histologically diagnosed with squamous cell carcinoma. Monoclonal antibodies to MMP-1 were used as primary molecular markers; MMP-9 as well as TIMP-1 and TIMP-2 (Termo Scientific, USA). In evaluating IHC reactions with the marker, the expression was calculated as the percentage of cells with a cytoplasmic reaction at increased (× 400) in 4 gradations: (1+) - <10%, (2+) - from 10% to 50%, (3+) - from 51% to 100%. The expression of TIMP1 and TIMP2 was assessed as negative with a percentage of stained cells <30%, “-” - 0, and > 30% as “+” - 1.
Results: Statistically significant difference in the expression of MMP-1 and MMP-9 between the groups in the presence of metastases (p <0.01) was revealed: patients with metastases were more likely to have moderate (++) and high (+++) expression of biomarkers, and rarely - low expression (+). Positive TIMP-2 was significantly more common among patients with metastases (p = 0.05).
MMP-1 and TIMP-2 were significant predictors of metastasis (Table 2). Increase in the expression of MMP-1 by one category increased the chances of metastasis almost by 5 times (OR = 4.75 (95% CI 1.77 – 14.75)). In positive TIMP-2, the chances of tumor metastasis were by 2.7 times higher (95% CI 1.01 – 7.89) than with negative TIMP-2. The difference between the ROC curves for MMP-1 and TIMP-2 was not statistically significant (p = 0.30). Expression of any of the indicators did not allow predicting the recurrence of the disease.
The inclusion of age in the regression model did not have a positive effect on the prognostic ability of predictors.
Conclusion: Expression rates of molecular markers MMP-1, MMP-9, TIMP-1 and TIMP-2 were statistically associated with the degree of tumor differentiation (p <0.05). According to the results of statistical data processing, MMP-1 and TIMP-2 were significant predictors of metastasis (p <0.01). Increase in the expression of MMP-1 to a positive or high level increased the chances of metastasis by 5 times (OR = 4.75 (95% CI 1.77 – 14.75). In positive TIMP-2 the chances of tumor metastasis were by 2.7 times higher) (95% CI 1.01 -7.89 ) than in the negative.