. . . "4"^^ . "2008-12-31+01:00"^^ . "1"^^ . " primary culture" . "1"^^ . . . . "2005-01-01+01:00"^^ . "C\u00EDlem projektu je funk\u010Dn\u00ED charakterizace exprese mutantn\u00EDch forem gen\u016F BRCA1 a BRCA2 v in vitro modelu prim\u00E1rn\u00EDch bun\u011Bk kultivovan\u00FDch ze \u017El\u00E1zov\u00E9 tk\u00E1n\u011B prsu nosi\u010Dek mutac\u00ED v genech BRCA1 nebo BRCA2. Po odb\u011Bru punk\u010Dn\u00ED biopsi\u00ED budou bu\u0148ky kultivov\u00E1ny s vyu\u017Eit\u00EDm %22feeder-layer%22 techniky. V bu\u0148k\u00E1ch in vitro bude exprese wt alely inhibov\u00E1na pomoc\u00ED RNA interference. Studov\u00E1na bude BRCA1/2-dependentn\u00ED exprese gen\u016F, resp. jej\u00ED alterace, pomoc\u00ED cDNA array. Charakterizace c\u00EDl\u016F defektn\u00ED signalizace BRCA1/2 bude kvantifikov\u00E1na pomoc\u00ED qRT-PCR a western blottingu; defektn\u00ED lokalizace genov\u00FDch produkt\u016F bude studov\u00E1na pomoc\u00ED separa\u010Dn\u00ED centrifugace. V prim\u00E1rn\u00EDch bu\u0148k\u00E1ch bude studov\u00E1n \u00FA\u010Dinek protin\u00E1dorov\u00FDch l\u00E9\u010Div v z\u00E1vislosti na diferencovan\u00E9 expresi mutantn\u00EDch resp. wt forem gen\u016F BRCA1/2." . "Experimentally modified gene expression of BRCA1 and BRCA2 variants in primary mammary cells in vitro: contribution to breast cancer development and sensitivity to diverse options of cancer therapy in BRCA1/2 genes mutation carriers"@en . . . . . . . " RNA interference" . "NR8345" . "BRCA1/2 mutation does not represetnt an important prognostic factor in cytotoxic breast cancer treatment."@en . "breast cancer; mutation; BRCA1; RNA interference; primary culture; heredity; BRCA2; cytostatics"@en . "2011-04-27+02:00"^^ . "2008-03-05+01:00"^^ . . " heredity" . "breast cancer" . . . "Mutace BRCA-1/2 nereprezentuje v\u00FDznamn\u00FD prediktivn\u00ED faktor pro cytostatickou l\u00E9\u010Dbu ca mmy"@cs . " BRCA2" . "Experiment\u00E1ln\u011B modifikovan\u00E1 exprese sekven\u010Dn\u00EDch variant gen\u016F BRCA1 a BRCA2 v prim\u00E1rn\u00EDch mam\u00E1rn\u00EDch bu\u0148k\u00E1ch in vitro: vliv na vznik karcinomu prsu a jeho ovlivn\u011Bn\u00ED r\u016Fzn\u00FDmi modalitami protin\u00E1dorov\u00E9 l\u00E9\u010Dby u nosi\u010D\u016F heredit\u00E1rn\u00EDch mutac\u00ED" . "The aim of the project is characterization of cellular changes produced by expression of mutant BRCA1 and BRCA2 genes in the in vitro model of mammary cells cultivated from breast glandular structures of BRCA1/2 mutation carriers. Following fine-needle biopsy, the cells will be cultivated using %22feeder layer technique%22. BRCA1/2 wt allele expression will be silenced using RNA interference. BRCA1/2-dependent gene expression will be studied using cDNA arrays. Selected targets of altered BRCA1/2 signalization will be quantified using qRT-PCR and western blotting; mislocalization of gene products will be fractionated using centrifugation. The effect of anticancer therapy will be studied in primary mammary cells expressing wt, or mutant, or both alleles of the BRCA1/2 specified gene alterations."@en . "http://www.isvav.cz/projectDetail.do?rowId=NR8345"^^ . . " mutation" . . " BRCA1" . "0"^^ . . "4"^^ .