Kudelski Jacek, Młynarczyk Grzegorz, Darewicz Barbara, Bruczko-Goralewska Marta, Romanowicz Lech
Department of Urology, Medical University of Białystok, Poland.
Department of Medical Biochemistry.
Medicine (Baltimore). 2020 Feb;99(7):e19224. doi: 10.1097/MD.0000000000019224.
Human urinary bladder cancer is one of the most common cancers worldwide with the mortality rate of approximately 165,000 people annually. The modulation of extracellular matrix is a crucial event in the metastatic spread, among others in angiogenesis. It is initiated and prolonged by the cascade of matrix metalloproteinases. MMP-14 and MMP-15 are associated with a high degree of malignancy, aggressiveness, and survival prognosis by the activation of other matrix metalloproteinases (MMPs). This study was aimed at evaluating the expression and the activity of selected transmembrane metalloproteinases at different stages of human urinary bladder cancer.
Western blot and enzyme linked immunosorbent assay (ELISA) method were used to evaluate the expression and content of MMPs and TIMP-1. The activity of studied enzymes was determined with fluorometric method.
Both transmembrane metalloproteinases are found in healthy or cancerous tissue in high molecular complexes of human urinary bladder. MMP-14 dominates over MMP-15, particularly in high-grade urinary bladder cancer. Their contents significantly change with the grade of bladder tumor. The amount of MMP-14 increases with increasing grade of tumor. MMP-15 content decreases in high-grade bladder cancer. With increasing grade of urinary bladder cancer their actual activity (per kg of total protein content) is varying in different ways. In all examined tissues, the specific activity of MMP-15 (per kg of the enzyme content) is much higher in comparison to MMP-14. Human urinary bladder cancer contains higher TIMP-1 amounts than control tissue but with the decrease with an increase in tumor grade.
Comparison of investigated enzymes' activity and the inhibitor content suggests it opposite effects, higher suppression of MMP-14 than MMP-15 activity in low-grade bladder cancer and reverse TIMP-1 action in high-grade cancer. The MMP-14 activity determination in urinary bladder cancer tissue may be used as a predictor of a risk of metastasis.
人类膀胱癌是全球最常见的癌症之一，每年死亡率约为16.5万人。细胞外基质的调节是转移扩散中的关键事件，尤其是在血管生成方面。它由基质金属蛋白酶级联反应启动并持续。MMP - 14和MMP - 15通过激活其他基质金属蛋白酶（MMPs）与高度恶性、侵袭性和生存预后相关。本研究旨在评估人类膀胱癌不同阶段所选跨膜金属蛋白酶的表达和活性。
采用蛋白质免疫印迹法和酶联免疫吸附测定（ELISA）法评估MMPs和TIMP - 1的表达及含量。用荧光法测定所研究酶的活性。
在人类膀胱的健康或癌组织中，两种跨膜金属蛋白酶均以高分子复合物形式存在。MMP - 14在MMP - 15中占主导地位，尤其是在高级别膀胱癌中。它们的含量随膀胱肿瘤分级显著变化。MMP - 14的量随肿瘤分级增加而增加。高级别膀胱癌中MMP - 15含量降低。随着膀胱癌分级增加，它们的实际活性（每千克总蛋白含量）以不同方式变化。在所有检测组织中，MMP - 15的比活性（每千克酶含量）相比MMP - 14要高得多。人类膀胱癌组织中TIMP - 1含量高于对照组织，但随肿瘤分级增加而降低。
所研究酶活性与抑制剂含量的比较表明其作用相反，在低级别膀胱癌中对MMP - 14活性的抑制高于MMP - 15，而在高级别癌症中TIMP - 1作用相反。膀胱癌组织中MMP - 14活性测定可作为转移风险的预测指标。
