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Universitatea de Medicina si Farmacie “Victor Babes”, Timisoara. Benteu Darius, seria E, grupa 34. Expresia crescuta a Translocazei 1 a Nucleotidului Adenina (ANT1) induce apoptoza si regresie tumorala in vivo. Apoptoza. Moarte celulara programata Dereglari – neoplazii, metastaze
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Universitatea de Medicina si Farmacie “Victor Babes”, Timisoara Benteu Darius, seria E, grupa 34
Expresia crescuta a Translocazei 1 a Nucleotidului Adenina (ANT1) induce apoptoza si regresie tumorala in vivo
Apoptoza • Moarte celulara programata • Dereglari – neoplazii, metastaze • Studiul genelor a caror expresie determina apoptoza
ANT-adenine nucleotide translocase • Schimb ATP-ADP=> ↑ ATPc • Exista 4 izoforme: • ↑ ANT1 => ↓ activitatii NFkB • NFkB – factor important in dezvoltarea tumorii • 1 – inima, muschi, creier • 2 – ficat
Obiective • Efectul anticancer al supraexpresiei ANT1 • Terapie genica pentru ANT1 in celulele canceroase in vivo si in vitro
Materiale • Linia celulara folosita: MDA-MB-231 • Soareci balb/c de 6-8 saptamani • Kituri RT-PCR/PCR • Plasmidul: pcDNA 3.1-ANT1
Metode in vitro • Celulele cultivate pe 6 placi • Transfectia utilizand plasmidul Examinarea celulelor: • Extractia DNA • Masurare potential membranar • Anticorpi anti NFkB
Metode – in vivo • 2 grupuri • A fost stabilit un model de tumora • Ziua 14: terapia pentru grupul de studiu • La fiecare 3 zile – tumora examinata
Rezultate • Statistica: student t, p<0,05 semnificativa • In vitro: ↑ANT1=> 30-40% celule distruse • In vivo: ↑ ANT1=> • Inhibarea cresterii tumorii • Diferente mari intre cele 2 grupuri • Ziua 40: tumora la jumatate
Concluzii • Cresterea tumorii a fost inhibata prin: • Inactivarea NFkB • Inducerea apoptozei • Metode mai eficiente pentru terapia genica in vivo
Referinte • 1. Gorczyca W, Melamed MR, Darzynkiewicz Z. Programmed death of cells (apoptosis) • 2. van Delft MF, Huang DC. How the Bcl-2 family of proteins interact to regulate apoptosis. • 3. Cotter TG, Lennon SV, Glynn JG, Martin SJ. Cell death via apoptosis and its relationship to growth, development and differentiation of both tumor and normal cells. • 4. Fulda S, Debatin KM. Modulation of apoptosis signaling for cancer therapy. • 5. Klingenberg M. Energetic aspects of transport of ADP and ATP through the mitochondrial membrane.