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Study of the function of ANKHD1 in apoptosis and its interaction with SIVA in cancer cells

Grant number: 09/00268-8
Support type:Regular Research Grants
Duration: April 01, 2009 - March 31, 2011
Field of knowledge:Health Sciences - Medicine - Medical Clinics
Principal Investigator:Fabíola Traina
Grantee:Fabíola Traina
Home Institution: Centro de Hematologia e Hemoterapia (HEMOCENTRO). Universidade Estadual de Campinas (UNICAMP). Campinas , SP, Brazil

Abstract

One step in the path towards building a comprehensive molecular portrait of human cancer is the definition of actively-expressed genes and the function of their coding proteins. The Ankyrin Repeat Single KH Domain containing 1 protein (ANKHD1) was first described in humans in a prostate carcinoma cell line LNCaP. We recently reported that ANKHD1 is located in the cytoplasm of cells and is overexpressed in human leukemia cell lines (KG-1, HEL, K562, NB4, HL-60, Jurkat, MOLT4, Raji, Daudi and Namalwa) and in 38 patients with a diagnosis of acute leukemia, compared to normal hematopoietic cells. Additionally, through the yeast two-hybrid system, using a cDNA library from normal human bone marrow, we detected an interaction between ANKHD1 and SIVA, a pro-apoptotic protein. These data suggest that ANKHD1 may be involved in apoptosis signalling in cancer cells. Thus, the aim of the present study is to confirm the interaction between ANKHD1 and SIVA and to investigate whether ANKHD1 is involved in apoptosis signalling in leukemia cells (Jurkat) and prostate carcinoma cells (LNCaP). To confirm the in vitro interaction between ANKHD1 and SIVA, both proteins will be transfected into HEK-293N cells, followed by immunoprecipitation and immunoblotting; to study the endogenous interaction, total extracts of Jurkat and LNCaP will be submitted to immunoprecipitation and immunoblotting with specific antibodies. ANKHD1 expression will be inhibited through RNAi and cells will be submitted to functional studies to evaluate the apoptosis signalling. (AU)

Scientific publications
(References retrieved automatically from Web of Science and SciELO through information on FAPESP grants and their corresponding numbers as mentioned in the publications by the authors)
MACHADO-NETO, JOAO AGOSTINHO; LAZARINI, MARIANA; FAVARO, PATRICIA; CAMPOS, PAULA DE MELO; SCOPIM-RIBEIRO, RENATA; FRANCHI JUNIOR, GILBERTO CARLOS; NOWILL, ALEXANDRE EDUARDO; MOURA LIMA, PAULO ROBERTO; COSTA, FERNANDO FERREIRA; BENICHOU, SERGE; OLALLA SAAD, SARA TERESINHA; TRAINA, FABIOLA. ANKHD1 silencing inhibits Stathmin 1 activity, cell proliferation and migration of leukemia cells. BIOCHIMICA ET BIOPHYSICA ACTA-MOLECULAR CELL RESEARCH, v. 1853, n. 3, p. 583-593, MAR 2015. Web of Science Citations: 10.

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