Effection Of HCMV Infection On Gap Junction Genes And Connexins In Glioma Cells

Human cytomegalovirus(HCMV)is belong to herpes virus of ?-subgroup.The infection of HCMV is very common in the normal population.A series of studies have shown that HCMV is closely related to the pathogenesis of glioma cells.Cells in the brain tissue,mainly by the cell gap junction communications(GJIC)pass some information,cell gap junction communication is the most direct way of communication signals in the body.In addition,GJIC between glioma cells is predicted to play a critical role in glioma invasion,and the glioma invasion is a difficult point in clinical practice.The expression and functional abnormalities of gap junction genes and connexins are associated with a variety of neurological disorders,the study of gap junction genes and connexins in the future will be the new direction and focus.This study intends to explore the effection of HCMV infection on gap junction genes and connexins in glioma cells,and to understand the role of connexins in tumor pathological mechanism.Respectively,cultured and used HCMV ADl69 strain(MOI = 1)were infected with primary glioma cell and U251 cell,and the morphological changes were observed with phase contract microscope.Morphological observations results showed that primary glioma cell and U251 cell became big and round and fused each other after HCMV infection.The expression of GJA8 and GJB1 in HCMV-infected primary glioma cell and U251 cell were detected by agarose gel electrophoresis.The results showed that GJA8 and GJB1 were involved in primary glioma cell and U251 cell can be expressed.Expression of the GJA8 and GJB1 after HCMV AD 169 infection primary glioma cell and U251 cell for various time periods(0h,12 h,24h and 48h)were detected by RT-PCR.RT-PCR results shown that the relative expression level of GJB1 was decreased in primary glioma cell(p?0.05),and the relative expression level of GJA8 was decreased after 24h(p?0.05),there was no significant change before 24 h in primary glioma cell(p? 0.05).The relative expression level of GJA8 and GJB1 was decreased in U251 cell(p?0.05).Western-Blot was used to detect the expression of Cx50 and Cx32 after HCMV AD 169 infection primary glioma cell and U251 cell for various time periods(0h,12 h,24h and 48h).The Western-Blot results shown that the relative expression levelof Cx32 was decreased in primary glioma cell(p?0.05),and the relative expression level of Cx50 was decreased after 24h(p?0.05),there was no significant change before 24 h in primary glioma cell(p ? 0.05).The relative expression level of Cx32 and Cx50 was decreased in U251 cell(p ? 0.05).The U251 cell migration was detected by Wound Healing Assay after HCMV infection.The results showed that the cell migration were enhanced after HCMV AD 169 infection.The results above indicated that the expression of gap junction genes and connexins were different after HCMV infection.This paper provides a theoretical foudation for the expression of gap junction genes and connexins in relation to the degree of malignancy of the tumor.