title: Nef from SIV(mac239) decreases proliferation and migration of adenoid-cystic carcinoma cells and inhibits angiogenesis.
abstract: The HIV/SIV accessory protein Nef is known to down-modulate cell surface receptors that are required for virus entry such as CD4, CCR5 and CXCR4 to block lethal viral superinfection of the infected cell. The chemokine receptor CXCR4 also plays an important role in promoting cell proliferation, metastasis and tumor angiogenesis. Therefore it was of interest to evaluate if Nef can down-regulate CXCR4 in tumor cells since this could affect these critical prognostic parameters. The CXCR4-expressing cell line ACC3 that was derived from a salivary gland adenoid cystic carcinoma (ACC) of the head and neck was transfected with Nef from SIV(mac239) and cell surface expression of the receptor was monitored by FACS analysis. Real time proliferation of cells was measured with the xCELLigence system (Roche, Mannheim, Germany). Cell migration was detected by an in vitro scratch assay. Similarly, COS-7 cells were co-transfected with CXCR4 and Nef and were treated as described for ACC3. In vitro tube formation was deployed to assess the effect of Nef on angiogenesis. siRNA was used for CXCR4 knockdown. Cell surface down-modulation of endogenous CXCR4 could be observed in ACC3 cells after Nef-transfection as well as in COS-7 cells after co-transfection of CXCR4 and Nef. Proliferation as well as migration of Nef-transfected ACC3 tumor cells appeared significantly reduced. In vitro tube formation was significantly lowered after Nef-transfection or CXCR4 knockdown with siRNA. SIV-Nef could serve as an interesting tool to study the biologic behavior of CXCR4-expressing tumors such as ACC. Deploying SIV-Nef thereby could help in the discovery of new therapeutic approaches for the treatment of ACC and other CXCR4-expressing tumors.
authors.affiliation: Department of Otorhinolaryngology, Head and Neck Surgery, University Hospital Giessen and Marburg, Campus Marburg, Baldingerstrasse, D-35033 Marburg, Germany.
authors.forename: Chengzhong
authors.initials: C
authors.lastname: Cai
authors.forename: Fiona R
authors.initials: FR
authors.lastname: Rodepeter
authors.forename: Annette
authors.initials: A
authors.lastname: Rossmann
authors.forename: Afshin
authors.initials: A
authors.lastname: Teymoortash
authors.forename: Jin-Seok
authors.initials: JS
authors.lastname: Lee
authors.forename: Karl
authors.initials: K
authors.lastname: Quint
authors.forename: Pietro
authors.initials: P
authors.lastname: Di Fazio
authors.forename: Matthias
authors.initials: M
authors.lastname: Ocker
authors.forename: Jochen A
authors.initials: JA
authors.lastname: Werner
authors.forename: Robert
authors.initials: R
authors.lastname: Mandic
mesh_terms: D000818:Animals; D016282:Aspartic Acid Endopeptidases / Q000494:pharmacology*; D019556:COS Cells / Q000378:metabolism; D003528:Carcinoma, Adenoid Cystic / Q000378:metabolism*; D045744:Cell Line, Tumor; D002465:Cell Movement; D049109:Cell Proliferation; D002522:Chlorocebus aethiops; D015536:Down-Regulation; D006801:Humans; D066298:In Vitro Techniques; D009389:Neovascularization, Pathologic; D034741:RNA, Small Interfering; D019718:Receptors, CXCR4 / Q000378:metabolism*; D012468:Salivary Gland Neoplasms / Q000378:metabolism*; D014162:Transfection; D054334:Viral Regulatory and Accessory Proteins / Q000502:physiology*
chemical_list: C429284:NEF protein, SIV; D034741:RNA, Small Interfering; D019718:Receptors, CXCR4; D054334:Viral Regulatory and Accessory Proteins; D016282:Aspartic Acid Endopeptidases; C077164:SIV(mac) proteinase