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Functional and experimental study of protein Mrck-b in colorectal adenocarcinomas with or without metastasis

Grant number: 13/21232-7
Support type:Scholarships in Brazil - Doctorate
Effective date (Start): March 01, 2014
Effective date (End): February 28, 2017
Field of knowledge:Health Sciences - Medicine
Principal Investigator:Rafael Malagoli Rocha
Grantee:Julio Cirullo Neto
Home Institution: A C Camargo Cancer Center. Fundação Antonio Prudente (FAP). São Paulo , SP, Brazil

Abstract

Introduction: Colorectal cancer (CRC) is a neoplasm of epithelial origin and covers tumors of the colon and rectum in men and women. It is a treatable and often curable when in the absence of extension to other organs. However, approximately 50% to 60% of patients diagnosed with CRC will develop metastatic, which are most commonly found in liver and lung. Objective: To evaluate the importance of proteins Mrck-² and Cdc42 in the process of cell migration and metastasis in colorectal carcinomas, using cell lines and experimental models. Methods: 60 cases of CRC who developed metastases and 60 cases who did not develop metastases are retrospectively selected from the files of Department of Pathology, Hospital AC Camargo and his paraffin material will be redeemed for performing IHC. IHC and IFL will be performed for markers anti-Mrck-² and anti-Cdc42, the reactions will be analyzed in an automated confocal equipment. The Western blot will be performed to confirm the specificity of all antibodies and quantify protein expression in cell cultures will be conducted to assess migration and invasion between the different groups of cells via assay techniques for transwell migration and wound healing. The shRNA and the overexpression plasmid will be used to decrease or increase the proteins expression in metastatic and non metastatic cells, and with this conditions, the cell migration will be assessed by testing the transwell. Mice will be selected for evaluation of metastasis to the depletion of Cdc42 and Mrck-B. The samples from the tumor bank, will be extracted RNAs to assess gene expression and statistical analysis will be performed in program Prism 5.2.1 and significance will be considered when p <0,01. (AU)