title: Role of human cytomegalovirus in the proliferation and invasion of extravillous cytotrophoblasts isolated from early placentae.
abstract: OBJECTIVE
  We investigated the role of human cytomegalovirus (HCMV) and its mechanism in extravillous cytotrophoblast (EVT) proliferation and invasion in vitro.
  
  
  METHODS
  Differential enzymatic digestion combined with gradient centrifugation, was used to isolate primary EVT from human chorionic villi collected from early placentae of healthy pregnant women. HCMV infection was determined by immunofluorescence staining of HCMVpp65 antigen expression. An MTT assay was used to examine the role of HCMV in the proliferation of EVT. Quantitative real-time polymerase chain reaction (qRT-PCR), immunocytochemical staining and Western blots were carried out in a control group (EVT) and a virus group (EVT+HCMV) to examine the expression of major genes and protein in TGF-β/Smad signaling pathways in EVT 48 h after inoculation with HCMV. An in vitro cell invasion assay was performed to analyze the influence of HCMV on EVT invasion.
  
  
  RESULTS
  HCMV significantly inhibited the proliferation of EVT 48 h after viral infection (P < 0.05). The expression of TGF-β1, Smad1, Smad2, Smad3, Smad4, and Smad5 genes was significantly increased (P < 0.05), but that of TGF-β2, TGF-β3, TGFβRI, TGFβRII, Smad7, MMP2, and MMP9 was significantly decreased in the virus group 48 h after HCMV infection (P < 0.05). Smad7, MMP-2 and MMP-9 protein levels were significantly decreased and the TGF-β1 protein level was significantly increased in infected EVT (all P < 0.05).
  
  
  CONCLUSIONS
  HCMV may act on multiple steps of the TGF-β/Smad signaling pathway to impede EVT proliferation and invasion.
authors.affiliation: Center for Reproductive Medicine, Shandong Provincial Hospital, Shandong UniversityJinan 250021, China; Department of Obstetrics and Gynecology, The Central Hospital of TaianTaian 271000, Shandong, China.
authors.forename: Tao
authors.initials: T
authors.lastname: Liu
authors.affiliation: The Information Center, The Central Hospital of Taian Taian 271000, Shandong, China.
authors.forename: Xiaofei
authors.initials: X
authors.lastname: Zheng
authors.affiliation: The Information Center, The Central Hospital of Taian Taian 271000, Shandong, China.
authors.forename: Qin
authors.initials: Q
authors.lastname: Li
authors.affiliation: Department of Obstetrics and Gynecology, The Third Affiliated Hospital of Guangzhou Medical University Guangzhou 510120, Guangdong, China.
authors.forename: Juanjuan
authors.initials: J
authors.lastname: Chen
authors.affiliation: Department of Obstetrics and Gynecology, The First Affiliated Hospital of Anhui Medical University Hefei 230022, Anhui, China.
authors.forename: Zongzhi
authors.initials: Z
authors.lastname: Yin
authors.affiliation: Department of Obstetrics and Gynecology, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology Wuhan 430030, China.
authors.forename: Juan
authors.initials: J
authors.lastname: Xiao
authors.affiliation: Department of Obstetrics and Gynecology, The International Peace Maternity and Child Health Hospital of China Welfare Institute Shanghai 20030, China.
authors.forename: Dandan
authors.initials: D
authors.lastname: Zhang
authors.affiliation: Department of Obstetrics and Gynecology, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology Wuhan 430030, China.
authors.forename: Wei
authors.initials: W
authors.lastname: Li
authors.affiliation: Department of Obstetrics and Gynecology, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology Wuhan 430030, China.
authors.forename: Yuan
authors.initials: Y
authors.lastname: Qiao
authors.affiliation: Department of Obstetrics and Gynecology, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology Wuhan 430030, China.
authors.forename: Suhua
authors.initials: S
authors.lastname: Chen
keywords: Human cytomegalovirus; Smad7; TGF-β1; extravillous cytotrophoblast; invasion; proliferation