Effects Of HCMV Infection On Autophagy Of Glioma Cells U87 And Its Significance

Objective:1. To detect the effects of Human cytomegalovirus(HCMV) infection in U87 glioma cell autophagy, and investigate the change of expression of microtubule-associated protein 1light chain 3(LC3) and the variation of autophagy body after HCMV infection.2. To detect the change of expression of Beclin1 autophagy autophagy-related gene Beclinl and its protein, then explored HCMV relations with glioma occurrence, development and significance.Materials and methods:1. After HCMV AD169(MOI = 5) infection in U87 glioma cell for various time periods(6h,12 h,24h, and 48h), phase contrast microscope was used to observe the change of cell morphologies.2. After HCMV infection in after U87 glioma cell for various time periods, electron microscope was used to observe the numbers and the morphologies of autophagosome.Western-blot and immunofluorescence were used to test the expression of LC3 protein.At the same time, the HCMV-uninfected cells in control groups were detected.3. After HCMV infection in U87 glioma cell for various time periods(6 h,12 h,24 h,and48 h), and uninfected-cells as a control group. RT-PCR was used to detect the change of expression of Beclin1, Western-blot and immunofluorescence were used to test the expression of Beclin1 protein. At the same time, the HCMV-uninfected cells in control groups were collected for detection.4. After HCMV infection in U87 glioma cell for various time periods, cell proliferation was detected by CCK 8 assay.Results:1. The morphology of cells were greatly different,after HCMV infection in U87 cells.2. The results showed that the numbers and the morphologies of autophagosome were changed. And compared with the control group, the expression level of LC3-II protein was gradually decreased in infected U87 glioma cell(P﹤0.05).3. Compared with the uninfected control group, the expression level of Beclin1 gene and protein was downregulated in infected U87 glioma cell(P﹤0.01).4. The proliferation activity of infected U87 glioma cell was distinctly elevated comparing with uninfected U87 glioma cell(P﹤0.01).Conclusions:1. HCMV infection blocks U87 glioma cell autophagy.2. HCMV infection restrain the expression level of Beclin1, then promote glioma cell proliferation.