| Nucleostemin (NS) is a newly discovered protein in embryonic stem cells and adult central nervous system stem cells, located in the nucleolus. differentiated cells and mature tissue cells have not been found. NS contains a basic domain, its function related to the localization in nucleolar, as well as the interaction with the P53under physiological conditions. NS plays an important role in cell cycle regulation in stem cells and tumor cells. FGF2(bFGF) was the first discovered angiogenesis factor, lots of studies have shown that FGF2can stimulate endothelial cell proliferation, migration, differentiation, and angiogenesis in vivo and in vitro:In addition, it is a mitogen. through the autocrine or paracrine method it can stimulate the growth of various tumor cells. It is overexpressed in a variety of tumor tissue. One of the method on analyse protein signaling networks is the study of protein interactions, the interaction between the protein molecules including molecular fusion of pre-hybridization, molecular recognition, molecular self-assembly, multi-molecular complexes, the energy charge transfer, these effects has important biological significance, these interactions are the molecular surface of the hydrophobic bond charge interactions between the molecules of these interactions. To detect these interactions we can not only calculate the condition of molecular surface charge and hydrophobic groups exposed, but also predict the intactions with the help of the data in some database. To study the indirect protein interactions, it can be observed by interfering with a protein’s expression then verifing another protein’s expression. Human esophageal cancer, especially esophageal squamous cell carcinoma has a high death rate in the world. In newly diagnosed patients with esophageal cancer in the world, more than1/2cases occurred in China. Esophageal cancer is involved in multi-factor, multi-stage diseases, including hyperplasia, dysplasia, carcinoma in situ and invasive cervical cancer, its mechanism of remains unclear. To explore the upstream and downstream proteins in the NS in signaling pathways, protein interaction networks, EC9706cells was choose as a carrier, using RNA interference to Interference NS’s and FGF2’s expression then its changes were detected by the Real-time PCR, and some protein interaction databases had been used to speculate the direct interaction. Thus its signaling pathway, upstream and downstream proteins-protein interaction networks can be predicted.Purpose Explore the node of NS in signaling pathways and protein interaction networks. Explore the possibilitiy ofb interaction etween NS and FGF2.Method EC9706cells was transfected with specific siRNA expression plasmid, to interfere with the expression of the NS and the expression of FGF2, Realtime PCR was used to detect the corresponding NS and FGF2expression. Use the database to predict the possibility of a direct interaction in NS and FGF2.Result Interfere with the expression of the NS, the NS relative expression level was0.04±0.02. FGF2relative expression of0.26±0.24:interfere with the expression of FGF2. the relative expression levels of the NS is0.78±0.27, the relative expression levels of FGF2was0.27±0.31; control group, the NS relative expression levels was1.05±0.14. FGF2relative expression was1.13±0.19. a statistically significant difference.(P<0.05). the results indicate that interfere with the expression of the NS will be lowered FGF2expression, interfere with the expression of FGF2at the same time was also lowered the expression of the NS. Database BioGrid3.0. STRING8.3databases, the PIR database’s results show the directly interactions between NS and FGF2Conclusion RNA interference and Realtime PCR experiments show that these two proteins in the indirect genetic level interaction. Database prediction results had been showen to suggested that there may be directly interactions between NS and FGF2. |
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