Over-expressed MIIP Gene Affected On HCMV-infected U87 Cells

Objective:1.To detect the change of expression of migration and invasion inhibitor protein(MIIP)after the infection of HCMV(human cytomegalovirus),and detect the change of the capacity of the migration and invasion inhibitor protein(MIIP)gen.2.In order to study the scientific value and the relationship between HCMV and glioblastoma,the change of the migration and invasion inhibitor protein(MIIP)gen and its expression and the fluctuation of the proliferation,invasion and migration of glioblastoma U87 were observed.Materials and methods:1 After HCMV AD169(MOI = 5)was infected for various time periods(6h,12 h,24h,48h)in U87 glioma cell,the change of cell morphologies was observed by phase contrast microscope.2 In this study,HCMV positive U87 cells infected with MIIP gene were used as experimental group,and U87 cells infected only with HCMV were used as control group,and HCMV infected U87 cells transfected by empty carrier CMV4 were used as empty plasmids.The expression of MIIP encoded protein was detected by Western-blot.3 CCK-8 was used to detect the proliferation of U87 cells.4 Tranwell migration test was employed to investigate the migration capacity of the U87 cells in the experimental group,the empty plasmid group and the control group.5 Tranwell invasion test and scratch test was applied to determine the invasion ability of the of the U87 cells in the experimental group,the empty plasmid group and the control group.Results:1 After infected with U87 cells,the morphology of HCMV cells changed significantly.2 The results of Western-blot indicated that the expression of MIIP gene in experiment group(U87+HCMV/MIIP)was much higher than those in empty plasmid group(U87+HCMV/CMV4)and blank group(U87+HCMV)(P <0.01).3 The results of CCK-8 test demonstrated that the proliferation capacity of cells inblank group was much higher than in experiment group(P <0.01).4 The results of Tranwell migration test demonstrated that both the numbers of the cells and the invasion capacity of cells decreased obviously in in experiment group,to compare with cells in blank group and empty plasmid group(P <0.01).5 The results of Tranwell migration test indicated that compared with blank group and empty plasmid group,the invasion ability of the cells in experiment group deduced significantly and the number of the cells decreased remarkably with P <0.01.In addition,the results of scratch test illustrated that the migration capacity of the transfected U87 in experiment group reduced significantly in comparison with that of the blank group and empty plasmid group.Conclusions:The infection of human cytomegalovirus(HCMV)population is very high and the virus of HCMV can appear alternately in the activation state and latent state.The latent infected virus can not produce and release new virus particles because of the inhibition of gene transcription,however,the new virus particles can be released under the active state of the virus.The activation of HCMV is affected by various kinds of factors such as transcription factors and cytokine.In addition,HCMV infection was potentially associated with the a variety of tumors.MIIP gene is a tumor over-expressed gene of newly discovered.Mitosis of tumor cells will out of control when the expression of MIIP gene is absent,and the cells are easy to form and migrate.In this study,changes of the characteristics of the U87 glioma cell were studied after the interference of over-expression of MIIP gene.The preliminary conclusions were drawn as follows:1.The proliferation activity of HCMV infected U87 cells was inhibited by the over-expressed MIIP gene.2.The migration capacity of HCMV infected U87 cells decreased after the overexpression of MIIP gene.3.The invasion ability of HCMV infected U87 cells was inhibited after the overexpression of MIIP gene.