Features of expression of immunohistochemical markers in local and generalized clear cell kidney cancer

Abstract

Purpose of the study. To conduct a comparative analysis of the expression of the following markers: Ki‑67, cyclin D1, E‑cadherin, CD44, MMP‑9, VEGF, P53, vimentin in renal tumor tissue in clear cell kidney cancer depending on the prevalence of the tumor process.Materials and methods. The material for the study was the operating material of 100 patients with light cell kidney cancer who were treated at the National Medical Reseaгch Centгe fог Oncology of the Russian Federation Health Ministry from 2015 to 2018. 50 patients were diagnosed with local cancer (T1–3an0m0), 50 – generalized cancer (T1–4N0M1). For the immunohistochemical (IHC) study, the material was fixed in 10% neutral formalin for 24 hours and encased in paraffin. Dewaxing and restoration of antigenic activity of the material was carried out in the RT module (Thermo Fisher Scientific) using Tris buffer pH=9, for 20 minutes at 98 °C. the Formulation of the IGC reaction was carried out in the immunohistostainer «Autostainer 480S» (Thermo Fisher Scientific). Used system detection UltraVision Quanto Detection System (Thermo Fisher Scientific), and the Chromogen DAB. Antibodies used: E‑cadherin (EP700Y) Cell Marque, 1: 100; CD 44 (EPR1013Y) Cell Marque 1:150; Ki‑67 (SP6) Spring Bio, 1:200; P53 (DO‑7) Cell Marque, 1:200; cyclin D1 (EP12) Dako, 1:200; VEGF Termo Fisher, 1:100; Vimentin (V9) Dako, 1:150; MMP‑9 (EP100902) Epitomics, 1:100. The results of the reactions with markers were evaluated by counting the number of colored cells in each 3rd field of view of the entire drug at magnification of the X200 lens in the AXIO Scope microscope. A1 (Carl Zeiss). The results were expressed as a percentage-the proportion of stained cells in relation to all tumor cells in the field of vision. Parametric methods of statistics were used for statistical processing of the results. The reliability of the difference between the two averages was determined by the student's t‑test for unrelated populations.Results. In clear cell kidney cancer, a low level of proliferative activity was observed in General, but in generalized, compared with local, it was significantly higher (PConclusion. Progression of clear cell kidney cancer from local to generalized forms is accompanied by hyperexpression of cyclin D1, a decreased e‑cadherin expression while increasing vimentin expression (increasing signs of epithelial-mesenchymal transformation), an increase in CD44 and VEGF expression.