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Mage-H1 and Mage-D1 regulation of the cell cycle and differentiation difference

Author: Ding
Tutor: LiuShaoJun
School: PLA Military Academy of Medical Sciences
Course: Cell Biology
Keywords: Mage-H1 Mage-D1 PC12 Cell cycle Cell differentiation
CLC: Q42
Type: Master's thesis
Year: 2008
Downloads: 26
Quote: 0
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The traditional view, neurons can not be split, but observed in the previous work of our laboratory, multiple sources of mature neurons can be split, and with continuous photographic methods to capture to in mitosis all time separatist neurons, further with a variety of biological technology, to prove that it has various features typical of mature neurons in the division neurons. This shows, the splitting of neurons in the nervous system of universal significance. Why neurons from the cell cycle? Why, under certain conditions, can be split? Explore the molecular mechanism of which have important biological significance. Start from the study of two structurally similar but the function is relatively molecular the Mage-H1 and Mage-D1, investigate their possible role in the cell cycle, the regulation of cell proliferation and differentiation, so as to explain the molecular mechanisms of neurons from the cell cycle lay the foundation. In-depth study, we first dominant epitopes Mage-H1 and Mage-D1 predict. Mage-D1 protein N-terminal 53aa-295aa prokaryotic expression and preparation for Mage-D1 protein antibody to lay the foundation for further study its function. (Mage-H1 antibody preparation section will be set forth in our laboratory a Master thesis.) This study found that, after the induction of differentiation of PC12 cells, Mage-H1 upregulation, at the same time, Mage -D1 expression was lowered. This shows that the Mage-H1 may be involved in the differentiation of PC12 cells, maintain cell resting midterm play an important function; Mage-D1 may promote the proliferation of PC12 cells maintain cell dedifferentiation state to play an important function. Subsequently, we will be able to express the plasmid with the GFP-tag Mage-H1 transfected PC12 cells and found that the part of the expression the Mage-H1 of PC12 cells differentiation. The flow cytometry analysis showed the Mage-H1 transient overexpression obvious G0-G1 phase arrest of PC12 cells. Western-blot detection PC12 cells after transfection Mage-H1 cDNA24hrs, Cdk2 that were down-regulated, and this downward can last 72hrs. Furthermore, we obtained were screened by G418 exogenous MAGE-H1 in PC12 cells stably expressing clones. Flow cytometry analysis showed that compared with the native PC12, PC12 cells stably expressing Mage-H1 clone obvious G0-G1 phase arrest. Moreover, Mage-H1 PC12 cells stably expressing clones were induced by NGF easier to differentiate into neurons. In the yeast two-hybrid screening experiments, we find the interacting proteins Rit2 of Mage-D1. Rit2 active form through ERK and p38 MAPK pathway to promote the differentiation of PC12 cells. We speculate that, Mage-D1 through inhibition of the activity of the Rit2 PC12 cells dedifferentiated state.

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