title: [Mycoplasma genitalium lipid-associated membrane proteins induce endocervical epithelial cells to express β-defensin-2].
abstract: OBJECTIVE
  To observe the expression of human β-defensin-2 (hBD-2) induced by Mycoplasma genitalium-derived lipid-associated membrane proteins (LAMPs) and its potential mechanism.
  
  
  METHODS
  Human endocervical epithelial End1/E6E7 cells were cultured in vitro and stimulated by different concentrations of LAMPs for 48 hours, and the expressions of hBD-2 mRNA and protein were detected by real-time RT-PCR and Western blotting, respectively. Toll-like receptor 2 (TLR2) and TLR6 neutralizing antibodies for End1/E6E7 cell cultivation, and dominant negative plasmids for cell transfection were used to analyze the roles of TLR2, TLR6 and MyD88 in mediating hBD-2 expression. Nuclear translocation of the nuclear factor κB (NF-κB) p65 subunit, and its DNA-binding activity were detected by Western blotting and electrophoretic mobility shift assay (EMSA), respectively. Pyrrolidine dithiocarbamate (PDTC), an inhibitor of NF-κB, was used to investigate the effect of NF-κB on hBD-2 expression.
  
  
  RESULTS
  Mycoplasma genitalium-derived LAMPs induced the expressions of hBD-2 mRNA and protein in End1/E6E7 cells. The expressions could be abrogated by TLR2 and TLR6 neutralizing antibodies, or their dominant negative plasmids. In addition, dominant negative plasmids of MyD88 significantly decreased LAMPs-induced hBD-2 expression. Western blotting showed that p65 was translocated to the nucleus, and the DNA binding activity of NF-κB was enhanced after LAMPs treatment. Furthermore, PDTC treatment decreased LAMPs- induced hBD-2 expression.
  
  
  CONCLUSIONS
  Mycoplasma genitalium-derived LAMPs can induce End1/E6E7 cells to express hBD-2, which may be involved in the TLR2, TLR6/Myd88/NF-κB pathways.
authors.affiliation: Departments of Obstetrics, Center Hospital of Hengyang, Hengyang 421001, China.
authors.forename: Jing
authors.initials: J
authors.lastname: Xie
authors.affiliation: Institution of Pathogenic Biology, Medical College, Hengyang 421001, China.
authors.forename: Xiaoxing
authors.initials: X
authors.lastname: You
authors.affiliation: Institution of Pathogenic Biology, Medical College, Hengyang 421001, China.
authors.forename: Yanhua
authors.initials: Y
authors.lastname: Zeng
authors.affiliation: Institution of Pathogenic Biology, Medical College, Hengyang 421001, China.
authors.forename: Ranhui
authors.initials: R
authors.lastname: Li
authors.affiliation: Institution of Pathogenic Biology, Medical College, Hengyang 421001, China.
authors.forename: Liesong
authors.initials: L
authors.lastname: Chen
authors.affiliation: Institution of Pathogenic Biology, Medical College, Hengyang 421001, China.
authors.forename: Cuiming
authors.initials: C
authors.lastname: Zhu
authors.affiliation: Institution of Pathogenic Biology, Medical College, Department of Clinical Laboratory, Affiliated Nanhua Hospital, University of South China, Hengyang 421001, China.
authors.forename: Jun
authors.initials: J
authors.lastname: He
authors.affiliation: Institution of Pathogenic Biology, Medical College, Hengyang 421001, China.
authors.forename: Minjun
authors.initials: M
authors.lastname: Yu
authors.affiliation: Institution of Pathogenic Biology, Medical College, Hengyang 421001, China.
authors.forename: Yimou
authors.initials: Y
authors.lastname: Wu
mesh_terms: D002478:Cells, Cultured; D002584:Cervix Uteri / Q000378:metabolism*; D004847:Epithelial Cells / Q000378:metabolism; D005260:Female; D006801:Humans; D058635:Lipid-Linked Proteins / Q000502:physiology*; D045704:Mycoplasma genitalium / Q000502:physiology*; D053594:Myeloid Differentiation Factor 88 / Q000502:physiology; D016328:NF-kappa B / Q000502:physiology; D051195:Toll-Like Receptor 2 / Q000502:physiology; D051216:Toll-Like Receptor 6 / Q000502:physiology; D023083:beta-Defensins / Q000235:genetics*
chemical_list: C106641:DEFB4A protein, human; D058635:Lipid-Linked Proteins; C507779:MYD88 protein, human; D053594:Myeloid Differentiation Factor 88; D016328:NF-kappa B; C495342:TLR2 protein, human; C495368:TLR6 protein, human; D051195:Toll-Like Receptor 2; D051216:Toll-Like Receptor 6; D023083:beta-Defensins