Prognostic value of a panel of immunohistochemical markers for determining the risk of recurrence and progression of non-invasive bladder cancer

Abstract

Background. There is a huge number of algorithms for the diagnosis and prognosis of the clinical course of non-invasive bladder cancer (NIBC). They are based on both clinical and morphological features and data from immunohistochemical (IHC) studies in which a large number of markers, including p53, Ki-67, VEGF, E-cadherin, MMP, bcl, etc., were used. Of scientific and practical interest is a comprehensive assessment of the prognostic value of a number of the main IHC markers used for the diagnosis and prognosis of bladder cancer, the impact of epithelial-mesenchymal transition (EMT) processes and the immune response on the risk of recurrence and tumor progression. Purpose. To study the prognostic value of the panel of IHC markers for determining the risk of recurrence and progression of NIBC. Materials and Methods. We studied surgically removed stage T1 malignant bladder tumors that spread to the submucosal tissue without invasion into the muscle tissue in 42 patients. Recurrence was understood as tumor development within 5 years after surgical treatment. The term «progression» meant the growth of the tumor in the muscle tissue, in other words, transformation of the tumor from non-muscle-invasive to muscle-invasive, which is of great prognostic significance for this type of cancer. IHC study was performed using primary ready-to-use monoclonal antibodies produced by DAKO (Denmark) for p53, p63, Ki67, E-cadherin, N-cadherin, CK7, CK20, Vimentin (VimentinV9), MMP-9, TGF- β1, VEGF, CD34+, CD3+, CD4+, CD8+, CD20+, CD68+, collagen IV. Epithelial markers (E-cadherin, cytokeratins 20 (CK20) and 7 (CK7)) and mesenchymal markers (N-cadherin and vimentin) were used to study EMT processes. Statistical processing of the research results was performed using the Statistica 6.0 package. Results and Discussion. It was found that the following markers may have prognostic value for the recurrence of NIBC: p53, CD3+, CD8+, CD68+; for recurrence with progression: Ki-67 and MMP-9. As differentiation of NIBC decreased, we observed increased expression of vimentin, Ki-67, MMP-9, VEGF, CD3+, CD8+, CD68+, p53, CD 34 and decreased expression of E-cadherin, CK20, collagen IV, p63. We established a relationship between the clinical behavior of NIBC and EMT processes. Thus, stage 1 EMT was characteristic of cancer with recurrence but without progression (p < 0.05), and stage 2 EMT was characteristic of NIBC with recurrence and progression (p < 0.05). The association of EMT with immune cell infiltration of NIBC was proven. The emergence of EMT in NIBC is associated with an increase in the intensity of infiltration by CD68+ macrophages (p < 0.01), CD3+-T-lymphocytes (p < 0.05), including both CD8+ (p < 0.01) and CD4+ (p < 0.05) cells. Conclusions. Non-progressive cancer with recurrence is characterized by stage 1 EMT of the tumor cells (p < 0.05), and progressive NIBC with recurrence is characterized by stage 2 EMT (p < 0.05). The association of EMT with immune cell infiltration of NIBC was proven. The emergence of EMT in NIBC is associated with an increase in the intensity of infiltration by CD68+ macrophages (p < 0.01), CD3+ T-lymphocytes (p < 0.05), including both CD8+ (p < 0.01) and CD4+ (p < 0.05) cells.