Malignant melanoma (MM) urgently needs identification of new markers with better predictive value than currently-used clinical and histological parameters. Cancer cells stimulate the formation of a specialized tumor microenvironment, which reciprocally affects uncontrolled proliferation and migration. However, this microenvironment is heterogeneous with different sub-compartments defined by their access to oxygen and nutrients. This study evaluated microvascular density (MVD), CD3+ lymphocytes (TILs) and FOXP3+ T-regulatory lymphocytes (Tregs) on formalin-fixed paraffin-embedded tissue sections using light microscopy. We analyzed 82 malignant melanomas, divided according to the AJCC TNM classification into four groups--pT1 (35), pT2 (17), pT3 (18) and pT4 (12)--and 25 benign pigmented nevi. All parameters were measured in both the central areas of tumors (C) and at their periphery (P). A marked increase in all parameters was found in melanomas compared to nevi (p = 0.0001). There was a positive correlation between MVD, TILs, FOXP3+ Tregs and the vertical growth phase. The results show that MVD, TILs and FOXP3+ Tregs substantially influence cutaneous melanoma microenvironment. We found significant topographic differences of the parameters between central areas of tumors and their boundaries.

CGB Laboratory a s Laboratory of Molecular Genetics and Pathology AGEL Research and Training Institute Ostrava Vitkovice Branch Korenskeho 10 Ostrava 71000 Czech Republic

CGB Laboratory a s Laboratory of Molecular Genetics and Pathology AGEL Research and Training Institute Ostrava Vitkovice Branch Korenskeho 10 Ostrava 71000 Czech Republic Department of Pathology Faculty of Medicine University of Ostrava Syllabova 19 Ostrava 70300 Czech Republic

Institute of Clinical and Molecular Pathology Faculty of Medicine and Dentistry Palacky University Olomouc Hnevotinska 3 Olomouc 77515 Czech Republic

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