Transmembranic glycoprotein E-cadherin interacts with the cytoskeleton via intracellular proteins
named catenins. Cell-cell cohesion can be damaged by the loss of E-cadherin expression or changes in catenin expression, which leads to the loss of cadherin function. The cadherin-catenin complex also influences migration and modifies cell growth and the survival of neoplastic cells . In addition, beta-catenin, a member of the catenin family, participates in signal transduction [16, 17]. There are no current immunohistochemical prognostic markers for RCCs in routine use. In this era of new treatment possibilities there remains a need for better prognostic tools to plan the treatment and follow-up of RCC patients. The purpose of this study was to examine for the first time the immunostaining of myosin VI in RCCs and to investigate the prognostic
potential of immunostaining Selleckchem BEZ235 myosin VI, E-cadherin and beta-catenin in RCCs. Methods Patients The study population has been described in detail earlier . Briefly, the retrospective study population consisted of 152 find more patients who underwent surgery for RCCs between 1990 and 1999 at the Oulu University Hospital in Finland. Seven patients (5%) were operated by resection and 145 (95%) by radical nephrectomy. The patients’ follow-up details were collected from patient records. Follow-up was completed in all cases. The research plan was approved by the local ethical board. The stage of the tumours was assigned using the TNM (tumour-node-metastasis) staging of RCCs .
Tumour samples The tumour samples were fixed in 10% buffered formalin and embedded in paraffin. Histological diagnosis was confirmed by reviewing haematoxylin and eosin (H & E)-stained original sections. The tumours Thiamet G were reclassified and graded according to the WHO classification . The most representative block was selected to reconstruct a multitissue block, which was used for immunohistochemistry. Immunostaining procedure The immunoexpression of myosin VI, E-cadherin and beta-catenin was analysed using monoclonal antibodies. The antibodies used in the study were monoclonal anti-myosin VI (Sigma, St. Louis, MO, USA) in a dilution of 1:250, mouse anti-E-cadherin (Zymed Laboratories, San Francisco, CA, USA) in a dilution of 1:300 and this website anti-beta-catenin (BD Biosciences, San Jose, CA, USA) in a dilution of 1:200. For antigen retrieval, the sections were incubated in 0.01 M citrate buffer (pH 6) twice for 5 min and boiled in a microwave oven to enhance immunoreactivity. The sections were cooled for 15 min in 0.05 M Tris buffered saline (TBS) (pH 7.5) and washed twice in PBS. Endogenous peroxidise activity was eliminated by incubation in 5% hydrogen peroxide and absolute methanol. Bound antibodies were visualised using an EnVision+ System-HRP (DakoCytomation, Glostrup, Denmark).