Charactrization Of Sema3C In Glioma

Objective: To investigate the regulation of Semaphorin3 c in gliomas.Methods: First,Bioinformatics software analysis and prediction the microRNA that may regulate the expression of Sema3C: 1,Input the gene name Sema3C in Gene Bank,in order to find its 3'UTR sequence.2,Using the Target Scan and miRan Da software,input the 3'UTR of Sema3C m RNA sequence and selection 5-10 candidate micro RNAs that may control the expression of Sema3C according to its score.3,To further narrow the scope of the target micro RNA to be reduced to 2-3.We use the Pub Med software for literature search to reach our aim.Second,Detectioned the regulation of micro RNA to Sema3C :1,Customized for human micro RNA mimics(mimics)and inhibitors.2,Human glioma cell lines(A172and U251)were cultured in vitro.Uses the methods of Lipotamine2000 transfection in order to makes the mimics and inhibitors of the target micro RNA and the corresponding control(mock control and inhibitor control)transfection into the above cell lines,and then we extraction the total protein and total RNA,Western blot and fluorescence quantitative PCR detection the expression of Sema3C.Third,determined whether the selected microRNA can affect the activity of Sema3C m RNA 3'UTR: 1,Extraction the total RNA of human glioma cell line A172,reverse transcription of c DNA and prediction the micro RNA binding sequester and then ligated it to the pmir GLo plasmid.2,Uses the methods of Lipotamine2000 transfection transfected the micro RNA mimics,inhibitors and their control into HEK293 T cells,the cell samples were detected after the detection of fluorescence signal values.Fourth,Collection of glioma tissue(about 30 cases)and peripheral normal brain tissue(about 30 cases),to do the following experiment:1,The total RNA and total protein were extracted,and detected the expression of Sema3C and micro RNA by Western blot and real-time PCR.2,Statistical analysis the expression of Sema3C and micro RNA in glioma and peripheral cancer tissue to detected whether they exist relevance.Results:1,The expression of Sema3C in histologic levelThe express of Sema3C in glioma(78.2%)was significantly higher than that in peripheral normal tissues(20.0%).The prognosis of patients with low expression of Sema3C was more better(p = 0.017).The expression of Sema3C in gliomas was significantly higher than that in peripheral tissues(P = 0.008)and pathological grade(p = 0.002).The mutant incidence of IDH1(soluble isocitrate dehydrogenase 1)was low(p = 0.0001)in the tissues with high expression of Sema3C;In addition,it was also associated with the Ki67 index(p = 0.02).2,The cytology function of Sema3C Silence Sema3C expression,the proliferation and invasion of cells significantly reduced;the ability to regulate proliferation is affected by the cell cycle to achieve;Sema3C can change the cell epithelial-mesenchymal transition affect the ability of cells and so on.3,Study the pathogenesis of Sema3C Sema3C is regulated by the expression of miR-142-5p in glioma cells.Mi R-142-5p is achieved by affecting the translational level of Sema3C m RNA.Mi R-142-5p also regulates the proliferation and invasion of glioma cells,And the effect of this action may also occur by affecting the cell epithelial-mesenchymal cell transformation.4,The correlation expression of Sema3C and miR-142-5p in histology level The expression of miR-142-5p in glioma tissue decreased with the increase of grade.The expression of Sema3C and miR-142-5p was negatively correlated in glioma tissue.Conclusion:1,Sema3C in glioma can be used as a poor prognostic indicators for clinical diagnosis continue to study;2,Sema3C in the glioma play the role of oncogene;The role of the its may be through the regulation of epithelial-stromal cell transformation and achieve;3,miR-142-5p negatively regulates the expression of Sema3C in gliomas,suggesting that miR-142-5p / Sema3 pathogenesis may exist during glioma lesions.