Analysis Of The Phosphoproteins Involved In Proliferation And Differentiation Stimulated By Retinoic Acid Of C17.2 Neural Stem Cells

Proteinphosphorylationisanimportantposttranslational modification.Protein phosphorylation is the most basic,common,and most important mechanism to adjust and control protein vitality and function.Reversiblephosphorylationregulatesthecellsignal transduction,differentiation,growth,apoptosis,almost all the cell life activities.Moreover,the protein phosphorylation disregulation will induce many human diseases such as cancer,alzheimer's disease,heart disease,diabetes.The crucial physiological significance and associated with a variety of serious disease makes the research on phosphoproteins has became a hot spot in proteomics studies.Neural stem cells?NSCs?are stem cells which have the capability of self-renewal and multi-directional differentiation potential.Under specific conditions they can differentiate into neurons,astrocytes and oligodendrocytes.NSCs'excellent biological features,have offered perspectives for treatment of damages and chronic degenerative diseases of the central nervous system.Brain realize its functions mainly through the nerve cells secreted neurotransmitters,discreted and conducted neuronal signaling.The key of NSCs transplantation achieve function reconstruction of the central nervous system and nerve regeneration is how to make the NSCs differentiate into functional neurons directionally.Researching the expression of phosphoproteins and the change of phosphorylated modification degree involved in the proliferation and differentiation of NSCs,it is benefit for deepen comprehension of the physiological mechanism of NSCs'proliferation and differentiation.It is expected to realize the directional differentiation of NSCs artificially,and lay the foundation for clinical NSCs transplantation therapy.C17.2 neural stem cells are the good carrier for the in vitro study of neural stem cells,because they have the qualities of immortality,multipotential,and could be tracked in vivo.There have been some researches show that retinoic acid can promote neural stem cells differentiate into neurons.Tumor metastasis is the main cause of malignant tumor death.Prostate cancer is a kind of epithelial malignant tumors happened in prostate.Prostate cancer cell can occur some kinds of metastasis to the other parts of the body,which create great harm to human healthy.The study of tumor metastasis mechanism will help to identify meaningful cancer treatment targets,it can lay the foundation for clinical diagnosis and treatment.Prostate cancer cell line Mat-LyLu is a material for the tumor metastasis study,veratrine is known to promote tumor metastasis,study the the phosphoproteins appear in the proliferation and metastasis of Mat-LyLu cell help us to find out the mechanism of tumor metastasis.This experiment adopts two biological models:C17.2 neural stem cells neuronal differentiation and prostate cancer Mat-LyLu cells metastasis,to study the phosphoproteins'dynamic changes in biological processes.After the total protein lysate trypsin digest in the solution,we used TiO₂ for the phosphorylated peptide concentration,and used BHD as the inhibitor of nonspecific adsorption.Do the nanoLC-ESI-MS mass spectrometry identification and bioinformatics analysis at last.In the C17.2 neural stem cell differentiation experiment,we identify 219 non redundant proteins,82 phosphoproteins,infer 366 phosphorylation sites according to the whole phosphorylated proteins sequence information.In the Mat-LyLu cell metastasis experiment,we found 279 non redundant proteins,69 phosphoproteins,infer 273 phosphorylation sites according to the whole phosphorylated proteins sequence information.These phosphoproteins play an important role in regulation of gene replication and transcription,metabolic,protein synthesis,molecular recognition and signal transmission.Our research enlarges the dataset of phosphoproteins of neural stem cells and prostate cancer cells,it is the foundation for the study of neural stem cell proliferation,directional differentiation,neural system disease treatment and prostate cancer cells metastasis mechanism,cancer therapy.