Effect Of Overexpressed Necdin On The Cell Cycle And Proliferation Of P19Embryonal Carcinoma Cells

Necdin is an important member in the type II melanoma antigen (MAGE) protein family, and mainly expressed in all dividing neurons in neural development process and mature neurons. As so far, studies have found that Necdin plays important roles in neural system:it can suppress cells proliferation through combining the transcription activity domain of E2F1; regulates cell apoptosis through interaction with p53; participates in the p75signaling pathways induced by NGF through interaction with p75to promote the neuronal differentiation; and is closely related with Prader-Willi syndrome, etc. As the study on Necdin progressed, detection improved. More and more close attention has been paying to the importance of Necdin. So we will study on the topic that the effect of overexpressed Necdin on the the cycle and proliferation of P19embryonal carcinoma cells based on excellent previous research in our laboratory as there has been no relevant report.In the study, we first constructed eukaryotic expression vectors and then prepared P19cell clones overexpressed exogenous Necdin stably which transfected with pCMV-tag2B-Necdin and PcDNA3.1-Necdin stably. The result assayed by WesternBlot showed that Necdin expressed in different level between the experimental group and the control group. We use flow cytometry and CCK-8assay respectively detected the difference of the cell cycle and proliferation between different cell groups.The study found that exogenous overexpression of Necdin in P19cells have no obviously effect on the cell cycle and proliferation of P19cells. But the specific mechanism of high expression of Necdin had no effect on P19cell cycle and proliferation still needs further exploration. This paper contains17pictures,4tables and50references.