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Description
  • The aim of the project is identification of BRCA1 splicing variants specificity regarding their significance as a prognostic factors and predictive markers of breast cancer. cDNA will be synthesized from RNA isolated from mammary tissue samples received from breast cancer patients and from women without breast cancer and from their peripheral blood. BRCA1 mRNA splicing variants will be identified using cDNA as a template for PCR amplification with primers localized in 5´ and 3´ BRCA1 UTR and subsequently analyzed by fragmentation gel electrophoresis (Agilent 2100). Detected BRCA1 splicing variants will be cloned into TA – cloning vector and sequenced. Tissue specific expression of particular variants will be quantified by qPCR. Differences between splicing patterns of lymphocytes and mammary tissue within patients with breast cancer and controls will be statistically evaluated. BRCA1 splicing variants within breast cancer patients will be evaluated in relation to disease course and treatment. (en)
  • Cílem projektu je identifikace tkáňové specifičnosti sestřihových forem genu BRCA1 s ohledem na jejich význam jako prognostických a prediktivních ukazatelů karcinomu prsu. Vzorky RNA izolované z peroperačně získané mamární tkáně pacientek s ca prsu a žen bez nádoru podstupujících mamoplastiku a izolované z periferní krve budou sloužit pro syntézu cDNA. Sestřihové varianty BRCA1 mRNA budou z cDNA amplifikovány s primery zasahujícími 5' a 3' UTR sekvece BRCA1 a analyzovány fragmentační gelovou elektroforézou (Agilent 2100). Alternativní sestřihové varianty budou zaklonovány pomocí TA-klonování do vektorů a sekvenačně charakterizovány ze získaných plasmidů. Tkáňová exprese jednotlivých variant BRCA1 bude kvantifikována pomocí qPCR. Statisticky budou vyhodnoceny rozdíly v sestřihovém vzorci buněk periferní krve a mamy a mezi pacientkami s ca prsu a nenádorovou populací, u pacientek s ca prsu budou vyhodnoceny sestřihové vzorce BRCA1 mRNA ve vztahu k průběhu a léčbě onemocnění.
Title
  • BRCA1 alternative splicing variants and their prognostic and predictive role in breast cancer (en)
  • Prognostický a prediktivní význam alternativních sestřihových variant genu BRCA1 u karcinomu prsu
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  • NT12280
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  • breast cancer; hereditary breast cancer; breast cancer predisposition; breast cancer predictive facto; BRCA1 gene; splicing; alternative splicing (en)
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  • breast cancer
  • BRCA1 gene
  • breast cancer predictive facto
  • breast cancer predisposition
  • hereditary breast cancer
  • splicing
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