1. Notch i Ikaros u diferencijaciji hematopoetskih stanica
- Author
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Konjević, Matea, Antica, Mariastefania, and Matulić, Maja
- Subjects
Hes1 ,Helios ,PRIRODNE ZNANOSTI. Biologija ,transkripcijski faktori ,semi-quantitative RT-PCR ,transcription factors ,višestruka lančana reakcija polimerazom ,semi-kvantitativni RT-PCR ,Notch ,Aiolos ,Hes ,NATURAL SCIENCES. Biology ,multiplex polymerase chain reaction - Abstract
Hematopoeza je proces nastajanja novih krvnih stanica. Transkripcijski faktori iz porodice Ikaros i puta dojavljivanja Notch reguliraju važne kontrolne točke tijekom hematopoeze koje će odlučiti u kojem će se smjeru stanica diferencirati. U radu je potvrđena i izmjerena ekspresija transkripcijskih faktora iz porodice Ikaros: Ikaros, Helios i Aiolos te puta dojavljivanja Notch: Notch1 i Hes1 u NB4 stanicama prije i poslije diferencijacije ATRA-om. U radu je uspješno uhodan, prilagođen i primijenjen protokol za metodu višestruke lančane reakcije polimerazom (multiplex PCR). Metodom je istovremeno umnoženo 6 odsječaka DNA u uzorku od 2500 stanica. Za multiplex PCR su odabrane početnice te provjerene i testirane na specifičnost i izbjegavanje sekundarnih struktura. Semi-kvantitativni RT-PCR je korišten u svrhu otkrivanja moguće promjene u razinama mRNA u stanicama prije i poslije diferencijacije. Smanjenje u ekspresiji gena Aiolos i Hes1 ukazuje na njihovu moguću ulogu u diferencijaciji i razvoju nezrelih mijeloidnih stanica. Hematopoiesis is the process of formation of new blood cells. Transcription factors from the Ikaros family and Notch signaling pathway regulate important checkpoints during hematopoiesis, which will decide in which direction the cells differentiate. In this work was confirmed and measured expression of transcription factors from the Ikaros family: Ikaros, Helios and Aiolos and Notch signaling pathway: Notch1 and Hes1 in NB4 cells before and after differentiation induced with ATRA. Protocol for the method of multiplex polymerase chain reaction (multiplex PCR) was successfully established, adapted and implemented. Six segments of DNA was simultaneously amplified in a sample of 2500 cells. For multiplex PCR, primers were selected and tested for specificity and to avoid secondary structures. Semi-quantitative RT-PCR was used to detect possible changes in the levels of mRNA in cells before and after differentiation. The reduction in expression of genes Aiolos and Hes1 suggests their possible role in differentiation and development of immature myeloid cells.
- Published
- 2012