BST-2 Protein Participates In The Proliferation And Migration Of Tumor Cells Induced By HCMV Infection

Objective:1.To investigate the proliferation,migration,and invasion of tumor cells after human cytomegalovirus(HCMV)infection of human glioma cells(U251/U87).2.The expression of IE1 and the expression of Bone Marrow Stromal cell antigen 2(BST-2)in human glioma cells infected with different concentrations of HCMV3.Detecting the phosphorylation level of HCMV-infected human glioma cells and the proliferation,migration and invasion of glioma cells by interference suppression of BST-2to investigate the effect of BST-2 on glioma cells infected with HCMV Play a role in the process.The effect of BST-2 expression on the proliferation,migration,and invasion of glioma cells was further studied.Methods:1.Different concentrations of HCMV(MOI=1/MOI=0.1)were infected with human glioma cells(U251/U87)to establish a virus-infected cell model.2.The effects of different concentrations of HCMV(MOI=1/MOI=0.1)infection on migration and invasion of human glioma cells(U251/U87)were examined by cell scratch healing assay and Transwell assay.3.Detection of cell proliferation after HCMV infection of human glioma cells(U251/U87)by Cell Counting Kit-8(CCK-8 kit).4.Use small interfering RNA(sh RNA)to inhibit the expression of BST-2 protein and screen for effective silencing targets.5.The effect of HCMV infection on the protein expression of IE1,BST-2,P-AKT was detected by western blot.6.Immunofluorescence labeling BST-2 detects BST-2 interference levels.7.Effect of flow cytometry on cell apoptosis after interference.8.Immunohistochemical staining for the expression of IE in different graded glioma cellsResults:1.After HCMV infection of human glioma cells(U251/U87),cell proliferation and migration were enhanced,and the promotion of high concentration of HCMV(MOI=1) was more obvious than that of low concentration of HCMV(MOI=1).2.After HCMV infection of human glioma cells(U251/U87),the early expression of virus proteins IE1 and BST-2 was significantly expressed,and was positively correlated with virus infection concentration and infection time.3.The si BST-2-171 and si BST-2-256 target sequences have a significant effect on the silencing of BST-2 after HCMV(MOI=1)infection of U251 cells.4.Human glioma cells(U251/U87)were infected or mock-infected with HCMV(MOI=1)for 48 h.After transfection with sh BST-2-171 and sh BST-2-256,they were inhibited by BST-2,and the markers were marked.The expression of fluorescent protein BST-2 and Western blot BST-2 were significantly decreased,and phosphorylation of Akt was significantly inhibited.Flow cytometry showed that interference with BST-2 had no significant effect on cell apoptosis.5.Human glioma cells(U251/U87)were infected or simulated with HCMV(MOI=1)for 48 h.Transfection with sh BST-2-171 and sh BST-2-256 interfered with BST-2inhibition.The proliferation and migration of tumor cells(U251/U87)were significantly reduced.Conlusion:1.Human glioma cells(U251/U87)infected with HCMV can increase their ability of proliferation and migration,and are positively correlated with virus infection concentration and infection time.2.BST-2 is involved in the proliferation and migration of malignant glioma cells caused by HCMV infection,and inhibition of BST-2 can make its proliferation and migration ability weaken.