The Role And Significance Of HCMV Infection In Epithelial - Mesenchymal Transition Of Cervical Cancer Siha Cells

Objective: 1. It aims to explore the effectiveness of human cytomegalovirus infection on epithelial-mesenchymal transition in cervical carcinoma Siha cells, meanwhile establishing model of epithelial-mesenchymal transition on cervical carcinoma Siha cells in vitro during HCMV and TGF-β1 synergistic action.2. To detect the change of expression and transcription level of protein E-cadherin, vimentin matrix metalloprotein-2 and its gene associated with epithelial-mesenchymal transition, then exploring the molecular biological mechanisms during cervical carcinoma Siha cells occurring epithelial-mesenchymal transition after HCMV infection.Materials and methods: 1 At first we cultured the cervical carcinoma Siha cell, while amplifying virus and detecting the viral titer, and divided them into the group of HCMV and TGF-β1 synergistic action and the group of TGF-β1 individual action.2 The experimental group was treated with 5 ng/ml TGF-β1 and HCMV AD169(MOI=5), meanwhile the control group was stimulated with TGF-β1 without HCMV infection. Phase contrast microscope was used to observe the change of cell morphologies at 24 h, 48 h, 72 h.3 After TGF-β1 and/or HCMV treated with Siha cell for various time points(0h, 24 h, 48 h and 72h), Western blot and RT-PCR were used to detect the change of transcription level of epithelial cell marker E-cadherin, mesenchymal cell marker vimentin and mediate molecular matrix metalloprotein-2.4 Cell proliferation rate and activity were detected by CCK-8 assay for various time points after HCMV infection in Siha cells.Results: 1 The results showed that the morphological change in experimental group happened on 48 h, which was earlier than observed on 72 h in control group.2 The transcription levels of epithelial cell marker E-cadherin in experimental group were significantly decreased afer TGF-β1 and HCMV synergisticly treated in Siha cell at 24 h、48h、72h,(P<0.01); while the transcription of vimentin in experimental group happened on 48 h, which was earlier than observed on 72 h in control group, meanwhile the transcription levels of mesenchymal cell marker vimentin in experimental group were up-regulated(P<0.01).3 The expression levels of E-cadherin in experimental group were significantly decreased afer TGF-β1 and HCMV synergistic action in Cervical carcinoma Siha cell at 24h、48h、72h(P<0.01); while the expression of vimentin and MMP-2 in experimental group happened on 48 h, which was earlier than observed on 72 h in control group, and the expression levels of vimentin and MMP-2 in experimental group were up-regulated(P<0.01).4 The proliferation activity of Siha cell in the experimental group was distinctly enhanced compared with control group(P<0.01).Conclusions: 1. HCMV and TGF-β1 synergistic action might promote the epithelial-mesenchymal transition in Cervical carcinoma Siha cell.2. Cervical carcinoma Siha cell occur epithelial-mesenchymal transition through the down-regulated expression of E-cadherin and the up-regulated expression of vimentin and MMP-2, which may promote the invasion and metastasis of cervical carcinoma.