title: [Effect of antibacterial peptide hCAP18/LL-37 on ovarian cancer microenvironment and the regulatory mechanism of its expression].
abstract: OBJECTIVE
  To investigate the effect of antibacterial peptide hCAP18/LL-37 on ovarian cancer microenvironment and the regulatory mechanism of its expression.
  
  
  METHODS
  We assessed the effect of macrophage-promoted ovarian cancer cells invasion using BioCoat Matrigel invasion chamber. The expressions of hCAP18/LL-37 and versican V1 were determined by real-time PCR and Western blot analysis. SKOV3 cells were transfected with shRNA plasmid to abrogate the expression of versican V1, and then the expression of hCAP18/LL-37 in macrophages and the invasiveness of SKOV3 cells were assayed.
  
  
  RESULTS
  The Matrigel invasion assay showed that after co-culture with macrophages for 4 days, the number of penetrated SKOV3 cells was 112.8±17.1/per high power field, significantly higher than that in the SKOV3 cells cultured alone (8.2±1.9/per high power field) (P<0.05). Addition of hCAP/LL-37 neutralizing antibody into the co-cultured macrophage-SKOV3 cells markedly inhibited the macrophage-promoted SKOV3 cells invasion. The penetrated SKOV3 cells was 22.2±5.6/per high power field, significantly lower than the 100.6±25.2/per high power field in the control macrophage- SKOV3 co-cultured cells (P<0.05). The expressions of hCAP18/LL-37 mRNA and protein in macrophages were remarkably enhanced upon co-culture with SKOV3 cells, but not changed in SKOV3 cells cultured alone. The expression and secretion of versican V1 in the ovarian cancer cells were also significantly increased after co-cultured with macrophages. Knockdown of versican V1 in SKOV3 cells by small interfering RNA significantly reduced the expression of hCAP18/LL-37 mRNA and protein in the macrophages, as well as decreased the invasiveness of SKOV3 cells (P<0.05).
  
  
  CONCLUSIONS
  In the cancer microenvironment, the macrophage-secreted hCAP18/LL-37 promote the invasiveness of ovarian cancer cells, and the hCAP18/LL-37 expression is regulated by versican V1 protein released by ovarian cancer cells.
authors.affiliation: Department of Clinical Laboratory, Tongji Hospital of Tonji University, Shanghai 200065, China.
authors.forename: Qian
authors.initials: Q
authors.lastname: Lu
authors.affiliation: Department of Clinical Laboratory, Tongji Hospital of Tonji University, Shanghai 200065, China.
authors.forename: Wenqiang
authors.initials: W
authors.lastname: Quan
authors.affiliation: Department of Clinical Laboratory, Tongji Hospital of Tonji University, Shanghai 200065, China.
authors.forename: Junlu
authors.initials: J
authors.lastname: Wu
authors.affiliation: Department of Clinical Laboratory, Tongji Hospital of Tonji University, Shanghai 200065, China.
authors.forename: Xian
authors.initials: X
authors.lastname: Zhang
authors.affiliation: Department of Clinical Laboratory, Tongji Hospital of Tonji University, Shanghai 200065, China.
authors.forename: Wei
authors.initials: W
authors.lastname: Ma
authors.affiliation: Department of Clinical Laboratory, Tongji Hospital of Tonji University, Shanghai 200065, China.
authors.forename: Li
authors.initials: L
authors.lastname: Pang
authors.affiliation: Department of Clinical Laboratory, Tongji Hospital of Tonji University, Shanghai 200065, China; Email: 186ld@163.com.
authors.forename: Dong
authors.initials: D
authors.lastname: Li
mesh_terms: D023181:Antimicrobial Cationic Peptides / Q000378:metabolism / Q000494:pharmacology*; D045744:Cell Line, Tumor; D018920:Coculture Techniques; D003094:Collagen; D004338:Drug Combinations; D005260:Female; D006801:Humans; D007797:Laminin; D008264:Macrophages / Q000378:metabolism; D009361:Neoplasm Invasiveness; D009363:Neoplasm Proteins / Q000378:metabolism*; D010051:Ovarian Neoplasms / Q000378:metabolism / Q000473:pathology* / Q000503:physiopathology; D010957:Plasmids; D011509:Proteoglycans; D012333:RNA, Messenger / Q000378:metabolism; D034741:RNA, Small Interfering; D060888:Real-Time Polymerase Chain Reaction; D014162:Transfection; D059016:Tumor Microenvironment / Q000187:drug effects*; D053675:Versicans / Q000378:metabolism*; D054804:Cathelicidins
chemical_list: D023181:Antimicrobial Cationic Peptides; D004338:Drug Combinations; D007797:Laminin; D009363:Neoplasm Proteins; D011509:Proteoglycans; D012333:RNA, Messenger; D034741:RNA, Small Interfering; C059485:matrigel; D053675:Versicans; D003094:Collagen; D054804:Cathelicidins